CN114303944B - Callus culture medium and extract of rosa tenuifolia, preparation method and application - Google Patents
Callus culture medium and extract of rosa tenuifolia, preparation method and application Download PDFInfo
- Publication number
- CN114303944B CN114303944B CN202011065238.4A CN202011065238A CN114303944B CN 114303944 B CN114303944 B CN 114303944B CN 202011065238 A CN202011065238 A CN 202011065238A CN 114303944 B CN114303944 B CN 114303944B
- Authority
- CN
- China
- Prior art keywords
- callus
- rosa
- tenuifolia
- callus proliferation
- rose
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 206010020649 Hyperkeratosis Diseases 0.000 title claims abstract description 187
- 239000001963 growth medium Substances 0.000 title claims abstract description 131
- 239000000284 extract Substances 0.000 title claims abstract description 115
- 235000011449 Rosa Nutrition 0.000 title claims abstract description 114
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims abstract description 64
- 241000220317 Rosa Species 0.000 claims abstract description 60
- GYDJEQRTZSCIOI-LJGSYFOKSA-N tranexamic acid Chemical compound NC[C@H]1CC[C@H](C(O)=O)CC1 GYDJEQRTZSCIOI-LJGSYFOKSA-N 0.000 claims abstract description 58
- 229960000401 tranexamic acid Drugs 0.000 claims abstract description 58
- KOGFZZYPPGQZFZ-QVAPDBTGSA-N (2s,3r,4s,5r)-2-(2-hydroxypropyl)oxane-3,4,5-triol Chemical compound CC(O)C[C@@H]1OC[C@@H](O)[C@H](O)[C@H]1O KOGFZZYPPGQZFZ-QVAPDBTGSA-N 0.000 claims abstract description 43
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 claims abstract description 32
- 229930003268 Vitamin C Natural products 0.000 claims abstract description 32
- 235000019154 vitamin C Nutrition 0.000 claims abstract description 32
- 239000011718 vitamin C Substances 0.000 claims abstract description 32
- 238000012258 culturing Methods 0.000 claims abstract description 16
- DFPAKSUCGFBDDF-ZQBYOMGUSA-N [14c]-nicotinamide Chemical compound N[14C](=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-ZQBYOMGUSA-N 0.000 claims abstract description 13
- 230000035755 proliferation Effects 0.000 claims description 80
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 claims description 74
- 239000002609 medium Substances 0.000 claims description 72
- 235000005152 nicotinamide Nutrition 0.000 claims description 37
- 229960003966 nicotinamide Drugs 0.000 claims description 37
- 239000011570 nicotinamide Substances 0.000 claims description 37
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 33
- 230000001954 sterilising effect Effects 0.000 claims description 17
- 238000004659 sterilization and disinfection Methods 0.000 claims description 15
- 229920001817 Agar Polymers 0.000 claims description 14
- 240000004153 Hibiscus sabdariffa Species 0.000 claims description 14
- 235000001018 Hibiscus sabdariffa Nutrition 0.000 claims description 14
- 229930006000 Sucrose Natural products 0.000 claims description 14
- 239000008272 agar Substances 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 14
- 239000005720 sucrose Substances 0.000 claims description 14
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 13
- 150000008442 polyphenolic compounds Chemical class 0.000 claims description 13
- 235000013824 polyphenols Nutrition 0.000 claims description 13
- 230000002829 reductive effect Effects 0.000 claims description 13
- 239000004480 active ingredient Substances 0.000 claims description 12
- 239000002537 cosmetic Substances 0.000 claims description 11
- 238000000227 grinding Methods 0.000 claims description 11
- 235000004789 Rosa xanthina Nutrition 0.000 claims description 10
- 239000007787 solid Substances 0.000 claims description 9
- 238000002137 ultrasound extraction Methods 0.000 claims description 9
- 229940088594 vitamin Drugs 0.000 claims description 9
- 229930003231 vitamin Natural products 0.000 claims description 9
- 235000013343 vitamin Nutrition 0.000 claims description 9
- 239000011782 vitamin Substances 0.000 claims description 9
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 9
- 230000002087 whitening effect Effects 0.000 claims description 9
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 8
- 229910052799 carbon Inorganic materials 0.000 claims description 8
- 238000005119 centrifugation Methods 0.000 claims description 7
- 239000000701 coagulant Substances 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- 229930195732 phytohormone Natural products 0.000 claims description 7
- 230000003712 anti-aging effect Effects 0.000 claims description 6
- 238000005286 illumination Methods 0.000 claims description 6
- 239000003963 antioxidant agent Substances 0.000 claims description 5
- 230000003078 antioxidant effect Effects 0.000 claims description 5
- 235000006708 antioxidants Nutrition 0.000 claims description 5
- 238000000605 extraction Methods 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- 239000008367 deionised water Substances 0.000 claims description 4
- 229910021641 deionized water Inorganic materials 0.000 claims description 4
- 238000003801 milling Methods 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 claims description 2
- 108010010803 Gelatin Proteins 0.000 claims description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 claims description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 2
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 239000008273 gelatin Substances 0.000 claims description 2
- 229920000159 gelatin Polymers 0.000 claims description 2
- 235000019322 gelatine Nutrition 0.000 claims description 2
- 235000011852 gelatine desserts Nutrition 0.000 claims description 2
- 239000008103 glucose Substances 0.000 claims description 2
- 239000012528 membrane Substances 0.000 claims description 2
- 239000011148 porous material Substances 0.000 claims description 2
- 238000009210 therapy by ultrasound Methods 0.000 claims description 2
- 241000109329 Rosa xanthina Species 0.000 claims 8
- 235000000659 Rosa rugosa Nutrition 0.000 claims 2
- 240000006066 Rosa rugosa Species 0.000 claims 2
- 125000000185 sucrose group Chemical group 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 143
- 230000000694 effects Effects 0.000 abstract description 36
- 210000004927 skin cell Anatomy 0.000 abstract description 20
- 238000004161 plant tissue culture Methods 0.000 abstract description 10
- 230000003064 anti-oxidating effect Effects 0.000 abstract description 5
- 230000008901 benefit Effects 0.000 abstract description 2
- 210000003491 skin Anatomy 0.000 description 37
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 16
- 210000002950 fibroblast Anatomy 0.000 description 15
- 102000003425 Tyrosinase Human genes 0.000 description 13
- 108060008724 Tyrosinase Proteins 0.000 description 13
- 241000282414 Homo sapiens Species 0.000 description 12
- FAIXYKHYOGVFKA-UHFFFAOYSA-N Kinetin Natural products N=1C=NC=2N=CNC=2C=1N(C)C1=CC=CO1 FAIXYKHYOGVFKA-UHFFFAOYSA-N 0.000 description 12
- QANMHLXAZMSUEX-UHFFFAOYSA-N kinetin Chemical compound N=1C=NC=2N=CNC=2C=1NCC1=CC=CO1 QANMHLXAZMSUEX-UHFFFAOYSA-N 0.000 description 12
- 229960001669 kinetin Drugs 0.000 description 12
- 230000001737 promoting effect Effects 0.000 description 12
- 230000032683 aging Effects 0.000 description 11
- 230000004792 oxidative damage Effects 0.000 description 11
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 8
- 230000001681 protective effect Effects 0.000 description 8
- 239000000523 sample Substances 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 7
- 230000015572 biosynthetic process Effects 0.000 description 7
- 230000002401 inhibitory effect Effects 0.000 description 7
- 210000002510 keratinocyte Anatomy 0.000 description 7
- 239000005631 2,4-Dichlorophenoxyacetic acid Substances 0.000 description 6
- 230000003833 cell viability Effects 0.000 description 6
- 230000006378 damage Effects 0.000 description 6
- 210000002615 epidermis Anatomy 0.000 description 6
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 description 6
- 230000005764 inhibitory process Effects 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 241000196324 Embryophyta Species 0.000 description 5
- 238000002835 absorbance Methods 0.000 description 5
- 229940079593 drug Drugs 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 229940123457 Free radical scavenger Drugs 0.000 description 4
- 230000007547 defect Effects 0.000 description 4
- 229940088598 enzyme Drugs 0.000 description 4
- 230000008099 melanin synthesis Effects 0.000 description 4
- 210000002752 melanocyte Anatomy 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000003647 oxidation Effects 0.000 description 4
- 238000007254 oxidation reaction Methods 0.000 description 4
- 230000036542 oxidative stress Effects 0.000 description 4
- 239000003375 plant hormone Substances 0.000 description 4
- 239000002516 radical scavenger Substances 0.000 description 4
- 239000012488 sample solution Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 102000008186 Collagen Human genes 0.000 description 3
- 108010035532 Collagen Proteins 0.000 description 3
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 3
- WTDRDQBEARUVNC-LURJTMIESA-N L-DOPA Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-LURJTMIESA-N 0.000 description 3
- 208000012641 Pigmentation disease Diseases 0.000 description 3
- 241000220222 Rosaceae Species 0.000 description 3
- 208000027418 Wounds and injury Diseases 0.000 description 3
- 230000005779 cell damage Effects 0.000 description 3
- 208000037887 cell injury Diseases 0.000 description 3
- 230000004663 cell proliferation Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 229920001436 collagen Polymers 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 229940074391 gallic acid Drugs 0.000 description 3
- 235000004515 gallic acid Nutrition 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 208000014674 injury Diseases 0.000 description 3
- 201000001441 melanoma Diseases 0.000 description 3
- 239000006870 ms-medium Substances 0.000 description 3
- 150000003254 radicals Chemical class 0.000 description 3
- 210000000434 stratum corneum Anatomy 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- PRPINYUDVPFIRX-UHFFFAOYSA-N 1-naphthaleneacetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CC=CC2=C1 PRPINYUDVPFIRX-UHFFFAOYSA-N 0.000 description 2
- -1 6-furfuryl methyl adenine Chemical compound 0.000 description 2
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 2
- 206010051246 Photodermatosis Diseases 0.000 description 2
- 235000000662 Rosa banksiae Nutrition 0.000 description 2
- 241001278830 Rosa banksiae Species 0.000 description 2
- 229930003270 Vitamin B Natural products 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- UQHKFADEQIVWID-UHFFFAOYSA-N cytokinin Natural products C1=NC=2C(NCC=C(CO)C)=NC=NC=2N1C1CC(O)C(CO)O1 UQHKFADEQIVWID-UHFFFAOYSA-N 0.000 description 2
- 239000004062 cytokinin Substances 0.000 description 2
- 210000004207 dermis Anatomy 0.000 description 2
- 210000001339 epidermal cell Anatomy 0.000 description 2
- 230000003203 everyday effect Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 229960000367 inositol Drugs 0.000 description 2
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 2
- 210000002780 melanosome Anatomy 0.000 description 2
- 230000008845 photoaging Effects 0.000 description 2
- 230000008832 photodamage Effects 0.000 description 2
- 230000019612 pigmentation Effects 0.000 description 2
- 230000008929 regeneration Effects 0.000 description 2
- 238000011069 regeneration method Methods 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 2
- 230000008591 skin barrier function Effects 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- 235000019156 vitamin B Nutrition 0.000 description 2
- 239000011720 vitamin B Substances 0.000 description 2
- OCZVHBZNPVABKX-UHFFFAOYSA-N 1,1-diphenyl-2-(2,4,6-trinitrophenyl)hydrazine;ethanol Chemical compound CCO.[O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1NN(C=1C=CC=CC=1)C1=CC=CC=C1 OCZVHBZNPVABKX-UHFFFAOYSA-N 0.000 description 1
- HXKWSTRRCHTUEC-UHFFFAOYSA-N 2,4-Dichlorophenoxyaceticacid Chemical compound OC(=O)C(Cl)OC1=CC=C(Cl)C=C1 HXKWSTRRCHTUEC-UHFFFAOYSA-N 0.000 description 1
- OVSKIKFHRZPJSS-DOMIDYPGSA-N 2-(2,4-dichlorophenoxy)acetic acid Chemical compound OC(=O)[14CH2]OC1=CC=C(Cl)C=C1Cl OVSKIKFHRZPJSS-DOMIDYPGSA-N 0.000 description 1
- CGIDKJRJBMFXKV-UHFFFAOYSA-N 6-n'-benzylpurine-6,6-diamine Chemical compound N1=CN=C2N=CN=C2C1(N)NCC1=CC=CC=C1 CGIDKJRJBMFXKV-UHFFFAOYSA-N 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- AHMIDUVKSGCHAU-UHFFFAOYSA-N Dopaquinone Natural products OC(=O)C(N)CC1=CC(=O)C(=O)C=C1 AHMIDUVKSGCHAU-UHFFFAOYSA-N 0.000 description 1
- 102000016942 Elastin Human genes 0.000 description 1
- 108010014258 Elastin Proteins 0.000 description 1
- 240000000731 Fagus sylvatica Species 0.000 description 1
- 235000010099 Fagus sylvatica Nutrition 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 229920002683 Glycosaminoglycan Polymers 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- WTDRDQBEARUVNC-UHFFFAOYSA-N L-Dopa Natural products OC(=O)C(N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-UHFFFAOYSA-N 0.000 description 1
- AHMIDUVKSGCHAU-LURJTMIESA-N L-dopaquinone Chemical compound [O-]C(=O)[C@@H]([NH3+])CC1=CC(=O)C(=O)C=C1 AHMIDUVKSGCHAU-LURJTMIESA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 244000141359 Malus pumila Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- 229940122791 Plasmin inhibitor Drugs 0.000 description 1
- 235000004469 Rosa graciliflora Nutrition 0.000 description 1
- 241000037110 Rosa graciliflora Species 0.000 description 1
- 230000037338 UVA radiation Effects 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 239000007640 basal medium Substances 0.000 description 1
- 210000002469 basement membrane Anatomy 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000005282 brightening Methods 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 230000002354 daily effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000002500 effect on skin Effects 0.000 description 1
- 229920002549 elastin Polymers 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 239000000686 essence Substances 0.000 description 1
- 238000005562 fading Methods 0.000 description 1
- 230000020764 fibrinolysis Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000007760 free radical scavenging Effects 0.000 description 1
- 230000003810 hyperpigmentation Effects 0.000 description 1
- 208000000069 hyperpigmentation Diseases 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical class C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 230000002934 lysing effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000003061 melanogenesis Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000000419 plant extract Substances 0.000 description 1
- 239000005648 plant growth regulator Substances 0.000 description 1
- 239000002806 plasmin inhibitor Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- ZUFQODAHGAHPFQ-UHFFFAOYSA-N pyridoxine hydrochloride Chemical compound Cl.CC1=NC=C(CO)C(CO)=C1O ZUFQODAHGAHPFQ-UHFFFAOYSA-N 0.000 description 1
- 229960004172 pyridoxine hydrochloride Drugs 0.000 description 1
- 235000019171 pyridoxine hydrochloride Nutrition 0.000 description 1
- 239000011764 pyridoxine hydrochloride Substances 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 230000011506 response to oxidative stress Effects 0.000 description 1
- 210000002374 sebum Anatomy 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000009759 skin aging Effects 0.000 description 1
- 210000001626 skin fibroblast Anatomy 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000011885 synergistic combination Substances 0.000 description 1
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 229960000344 thiamine hydrochloride Drugs 0.000 description 1
- 235000019190 thiamine hydrochloride Nutrition 0.000 description 1
- 239000011747 thiamine hydrochloride Substances 0.000 description 1
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
- 239000003104 tissue culture media Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
- 125000000969 xylosyl group Chemical class C1([C@H](O)[C@@H](O)[C@H](O)CO1)* 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/002—Culture media for tissue culture
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/52—Stabilizers
- A61K2800/522—Antioxidants; Radical scavengers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Birds (AREA)
- Developmental Biology & Embryology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Tropical Medicine & Parasitology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Cosmetics (AREA)
Abstract
The invention discloses a callus culture medium and extract of rosa tenuifolia, a preparation method and application. The callus culture medium of the rosa tenuifolia comprises the following components: basic culture medium for plant tissue culture, 1-50 mg/mL of vitamin C, 1-50 mg/mL of nicotinamide, 1-100 mg/mL of tranexamic acid and 0.002-0.1% of hydroxypropyl tetrahydropyran triol. Compared with the existing culture medium, the fine-stalk rose callus culture medium has the advantage that the effect of the fine-stalk rose callus cell extract obtained by culturing fine-stalk rose callus on the antioxidation of skin cells is obviously improved.
Description
Technical Field
The invention relates to a callus culture medium and extract of rosa tenuifolia, a preparation method and application thereof.
Background
Plant callus cells are widely applied to the industries of foods, medicines and cosmetics due to the abundant secondary metabolites, naturalness, environmental friendliness and controllable conditions. Taxol, an anticancer component obtained by using plant cell culture technology, was approved by FDA in 2000, and apple stem cells were used by many cosmetic brands for their cell protection function. The plant extracts of Rosaceae are used in foods and cosmetics, and have effects of improving skin color, regulating water-oil balance, and enhancing skin activity. The rosa tenuifolia (Rosa graciliflora Red. Et Wils) is a scarce rosaceae plant resource, is mainly distributed in the partial areas of Yunnan, sichuan, tibet and Shaanxi, and the roots and fruits of the rosa tenuifolia can be used as medicines, have the effects of improving immunity, converging and detumescence, and are rosa tenuifolia which is worthy of attention and development.
In recent years, various documents report that vitamin C, nicotinamide, tranexamic acid and hydroxypropyl tetrahydropyran triol have the effects of whitening skin, protecting skin cells, brightening skin color and the like, and are widely applied to cosmetic products. Studies show that vitamin C has the functions of resisting oxidation and assisting collagen synthesis in human skin, and can resist aging, reduce wrinkles, inhibit melanin generation and the like. The nicotinamide has the effects of inhibiting skin surface pigmentation and whitening skin, and the whitening mechanism is possibly related to the inhibition of transfer of melanosomes from melanocytes to keratinocytes, so that the nicotinamide can reduce percutaneous moisture loss, reduce sebum secretion, enhance skin barrier function, increase the cohesiveness and thickness of stratum corneum and increase the maturity of the stratum corneum when the nicotinamide is externally used. Tranexamic acid, also known as tranexamic acid, is a plasmin inhibitor that is associated with abnormal fibrinolysis and is able to reduce blood loss, in theory any damage to the formation of the stratum corneum may cause hyperpigmentation by the PA activation system, whereas tranexamic acid may inhibit the synthesis of melanin by inhibiting PA. Hydroxypropyl tetrahydropyran triol is a derivative of xylose extracted and separated from beech tree, and has the main effects of promoting synthesis of mucopolysaccharide and proteoglycan, promoting synthesis of collagen, restoring dermis-epidermis connection in aged skin, helping dermis maintain elasticity and promoting regeneration of damaged tissues. The functional components of the cosmetics are widely applied in the cosmetic industry at home and abroad, and are popular in recent years. However, no report on the use of the components for callus culture is available at home and abroad.
Disclosure of Invention
The invention aims to overcome the defect that the skin care effect of a cell extract obtained by culturing the callus of rosa tenuifolia by using a culture medium in the prior art is to be improved, and provides a callus culture medium of rosa tenuifolia, an extract, a preparation method and application. Compared with the existing culture medium, the culture medium for the fine-stalk rosa calluses provided by the invention has the advantage that the effect of the fine-stalk rosa calluses cell extract obtained by culturing the fine-stalk rosa calluses on the skin cell antioxidation is obviously improved.
The invention solves the technical problems by the following scheme:
a roselle callus culture medium comprising the following components:
basic culture medium for plant tissue culture,
Vitamin C1-50 mg/L,
1-50 mg/L nicotinamide,
1-100 mg/L of tranexamic acid and
and 0.002-0.1% of hydroxypropyl tetrahydropyran triol, wherein the percentage refers to the mass percentage of the hydroxypropyl tetrahydropyran triol in the fine-stalk rose callus culture medium.
Preferably, the concentration of vitamin C in the callus culture medium of Rosa tenuifolia is 3-45 mg/L, for example 10-20mg/L, further for example 5, 8, 10, 12, 14, 15, 20, 25, 30, 35 or 40mg/L; more preferably 8 to 12mg/L, for example 10mg/L.
Preferably, the concentration of said nicotinamide in said callus medium of Rosa tenuifolia is 3-45 mg/L, for example 10-20mg/L, further for example 5, 8, 10, 15, 20, 25, 30, 35 or 40mg/L; more preferably 8 to 12mg/L, for example 10mg/L.
Preferably, the concentration of tranexamic acid in the callus culture medium of the fine rose is 3-95 mg/L, for example 18-60 mg/L; for example, 10, 20, 30, 40, 50, 60, 70, 80 or 90mg/L; more preferably 18 to 22mg/L, for example 20mg/L.
Preferably, the hydroxypropyl tetrahydropyran triol accounts for 0.002-0.09% of the mass of the fine rose callus culture medium, for example 0.002-0.005%, and further for example 0.003, 0.005, 0.006, 0.007, 0.008 or 0.009. More preferably 0.002 to 0.003%, for example 0.002%.
Preferably, the mass ratio of the vitamin C to the nicotinamide to the tranexamic acid is: 1:1:2.
preferably, the mass concentration of the vitamin C, the nicotinamide and the tranexamic acid in the fine-stalk rose callus culture medium is 10mg/L, 10mg/L and 20mg/L respectively.
Preferably, the mass concentration of the vitamin C, the nicotinamide, the tranexamic acid and the hydroxypropyl tetrahydropyran triol in the fine-stalk rose callus culture medium is respectively 10mg/L, 20mg/L and 0.002-0.1%.
In the present invention, the fine-stalk rosa is generally a fine-stalk rosa (rosa. Gratiflora) plant of rosa genus of rosaceae family.
In the present invention, the basal medium may be selected from a medium conventionally used in plant tissue culture in the art, such as MS medium, N6 medium, B5 medium, SH medium or White medium, preferably MS medium.
Preferably, the fine-stalk rose callus culture medium further comprises glacial water, and the water source of the glacial water is preferably Himalayan mountain, such as the middle north slope of the Himalayan mountain; the water intake of glacier water is preferably day karst Bacounty Qu Dengni MABingchuan.
In the invention, the rosa tenuifolia callus culture medium can also comprise phytohormone according to the routine in the field.
The plant hormone may be selected from plant hormones conventional in the art, preferably from one or more of α -NAA, 6-BA, 2,4-D and KT; more preferably, 2,4-D and KT.
Wherein, the alpha-NAA refers to alpha-naphthylacetic acid, which is a plant growth regulator conventional in the field.
Wherein, 6-BA refers to 6-benzylaminoadenine, which is a cytokinin synthesized by man in the art.
Wherein, the 2,4-D refers to 2, 4-dichlorophenoxyacetic acid, which is a synthetic auxin analogue conventional in the art.
Wherein KT refers to kinetin, which is an endogenous cytokinin and has a chemical name of 6-furfuryl amino purine (or 6-furfuryl methyl adenine).
Preferably, the concentration of the plant hormone in the callus culture medium of the rosa tenuifolia is 3.0-4.0 mg/L, more preferably 3.5mg/L.
When the plant hormone is 2,4-D and KT, the concentration of 2,4-D in the fine-stalk rose callus culture medium is preferably 3mg/L, and the concentration of KT in the fine-stalk rose callus culture medium is preferably 0.5mg/L.
The pH of the callus culture medium of Rosa tenuifolia according to the present invention may be pH of a medium conventionally used in the art for plant tissue culture, for example, 5 to 7, preferably 5.8.
The callus culture medium of Rosa tenuifolia according to the present invention may further comprise a carbon source such as one or more of sucrose, maltose and glucose, preferably sucrose, according to the conventional culture medium requirements in the art. The concentration of the carbon source may be a concentration of a carbon source in a medium conventional in the art, for example, a concentration of 28 to 35g/L, preferably 30g/L in the callus culture medium of Rosa tenuifolia.
The callus culture medium of Rosa tenuifolia according to the present invention may further comprise a coagulating agent, such as agar and/or gelatin, preferably agar, according to the conventional culture medium requirements in the art. The concentration of the coagulant may be a concentration conventional in the art, for example, a concentration of 6 to 8g/L, preferably 7g/L in the callus culture medium of Rosa tenuifolia.
In a preferred embodiment of the invention, the callus culture medium of Rosa tenuifolia comprises the following components: MS, 2, 4-D3.0 mg/L, KT 0.5.5 mg/L, sucrose 30g/L, agar 7g/L, glacial water, vitamin C10mg/L, nicotinamide 10mg/L, tranexamic acid 20mg/L, and hydroxypropyl tetrahydropyran triol 0.002%. Preferably, the fine-stalk rose callus culture medium consists of the following components: MS, 2, 4-D3.0 mg/L, KT 0.5.5 mg/L, sucrose 30g/L, agar 7g/L, glacial water, vitamin C10mg/L, nicotinamide 10mg/L, tranexamic acid 20mg/L, and hydroxypropyl tetrahydropyran triol 0.002%.
The invention also overcomes the defect that the proliferation rate of the fine-stalk rose callus cultured by the culture medium in the prior art is not high enough and the skin care effect of resisting UVB is to be improved, and provides the fine-stalk rose callus culture medium which can obviously improve the proliferation rate of the fine-stalk rose callus, promote the growth of the fine-stalk rose callus and obviously improve the effect of resisting UVB on skin cells (HaCaT cells and FB cells).
The invention solves the technical problems by the following scheme: a roselle callus culture medium comprising the following components: a basal culture medium for plant tissue culture and 45-55 mg/L of vitamin C.
Wherein the concentration of vitamin C is preferably 50mg/L.
Wherein, the fine-stalk rose callus culture medium is preferably composed of the following components: a basal culture medium for plant tissue culture and 45-55 mg/L of vitamin C. More preferably, the concentration of the vitamin C in the callus culture medium of the rosa tenuifolia is 50mg/L.
The invention also overcomes the defect that the callus cell extract of the rosa tenuifolia cultivated by the culture medium in the prior art has the effect of promoting the proliferation of skin cells and the antioxidation of the skin to be improved, and provides the callus culture medium of the rosa tenuifolia, which can obviously promote the proliferation of the skin cells and the antioxidation of the skin.
The invention solves the technical problems by the following scheme: a roselle callus culture medium comprising the following components: basic culture medium for plant tissue culture and 90-110 mg/L of tranexamic acid.
Wherein, the tranexamic acid is preferably 100mg/L.
Wherein, the fine-stalk rose callus culture medium is preferably composed of the following components: basic culture medium for plant tissue culture and 90-110 mg/L of tranexamic acid. More preferably, the concentration of the tranexamic acid in the callus culture medium of the rosa tenuifolia is 100mg/L.
The invention also overcomes the defect that the cell extract obtained from the callus tissue of the rosa tenuifolia cultivated by the culture medium in the prior art has insufficient proliferation and tyrosinase inhibition rate on skin cells, and provides the callus tissue culture medium of the rosa tenuifolia, which has obvious effects on proliferation and tyrosinase inhibition on skin cells (HaCaT cells and FB cells).
The invention solves the technical problems by the following scheme: a roselle callus culture medium comprising the following components: 0.009-0.015% of a basic culture medium for plant tissue culture.
Wherein the concentration of the hydroxypropyl tetrahydropyran triol is preferably 0.01%.
Wherein, the fine-stalk rose callus culture medium is preferably composed of the following components: 0.009-0.015% of a basic culture medium for plant tissue culture. More preferably, the concentration of the hydroxypropyl tetrahydropyran triol is 0.01%.
Further, the invention also provides a preparation method of the rose-stalk callus culture medium, which comprises the following steps: mixing the components of the fine rose callus culture medium.
Wherein, the preparation method of the fine-stalk rose callus culture medium can also comprise a sterilization step according to the routine in the field, such as high-temperature sterilization.
When the callus culture medium of the rosa tenuifolia comprises vitamin C, the method preferably further comprises the step of adding the vitamin C after the temperature is reduced to below 65 ℃ after high-temperature sterilization;
alternatively, when the callus medium of Rosa tenuifolia comprises nicotinamide, it is preferable to add said nicotinamide after the temperature is reduced to below 65 ℃ after high temperature sterilization;
or when the callus culture medium of the rosa tenuifolia comprises tranexamic acid, the method preferably further comprises the step of adding the tranexamic acid after the temperature is reduced to below 65 ℃ after high-temperature sterilization;
or, when the fine-stalk rose callus culture medium comprises hydroxypropyl tetrahydropyran triol, preferably, after high-temperature sterilization, the hydroxypropyl tetrahydropyran triol is added after the temperature is reduced to below 65 ℃;
alternatively, when the callus culture medium of Rosa tenuifolia comprises vitamin C, nicotinamide, tranexamic acid and hydroxypropyl tetrahydropyran triol, it is preferable to add the vitamin C, nicotinamide, tranexamic acid and hydroxypropyl tetrahydropyran triol after the temperature is reduced to 65 ℃ or lower after high temperature sterilization.
The invention further provides a fine-stalk rosa calluses cell extract, which is obtained by extracting the cultured fine-stalk rosa calluses cells, wherein the cultured fine-stalk rosa calluses cells are obtained by culturing the fine-stalk rosa calluses by the fine-stalk rosa calluses culture medium.
Wherein, the conditions for culturing can be conventional plant callus culture conditions, preferably: the temperature is 20+/-1 ℃, the humidity is 60% -75%, the illumination intensity is 1500 Lx-2000 Lx, and the illumination is carried out for 12 hours every day.
The time of the cultivation may be a cultivation time conventional in the art, preferably 20 days.
Preferably, the solid content of the extract of the callus cells of Rosa tenuifolia is 3-5 mg/mL, more preferably 3.0-4.5 mg/mL, for example 3.26, 3.77, 3.98 or 4.07mg/mL. The solid content of the fine-stalk rosa calluses cell extract refers to the mass percentage of the mass of the rest part of the fine-stalk rosa calluses cell extract after drying.
The content of polyphenols in the extract of callus cells of Rosa tenuifolia is preferably 3.0-4.5%, for example 3.4, 3.5, 4.0 or 4.1%. The content of the polyphenol is the mass percentage of the polyphenol in the extract of the calluses of the rosa tenuifolia.
The content of Vc in the fine-stalk rosa-banksiae callus cell extract is preferably 1-2 mg/kg, for example 1.1, 1.2, 1.5 or 1.7mg/kg, and the content of Vc is the mass of Vc in each 1kg of fine-stalk rosa-banksiae callus cell extract based on the wet weight of cells.
The nicotinamide content of the callus cell extract of Rosa tenuifolia is preferably 100-600 mg/kg, for example 117.6, 200.8 or 518.6mg/kg. The content of the nicotinamide is the mass of the nicotinamide in each 1kg of the callus cell extract of the rosa tenuifolia based on the wet weight of cells.
The content of tranexamic acid in the callus cell extract of Rosa tenuifolia is preferably 100-500 mg/kg, for example 120, 216 or 518mg/kg. The content of the tranexamic acid is the mass of the tranexamic acid in each 1kg of the callus cell extract of the rosa tenuifolia based on the wet weight of cells.
The extraction method may be any extraction method conventional in the art. Such as ultrasonic extraction. Preferably, the method comprises the following steps: taking fresh cultured fine-stalk rosa calluses, grinding, adding a proper amount of water, performing ultrasonic treatment, centrifuging, taking supernatant, and filtering to obtain the fine-stalk rosa calluses extract.
Wherein the rate of grinding may be a grinding rate conventional in the art, such as 50 to 70Hz, for example 60Hz; the milling time may be a milling time conventional in the art, for example, 50 to 70 seconds, for example, 60 seconds; the number of grinding times may be a number of grinding times conventional in the art, for example, 1 to 3 times, and further, for example, 2 times.
Wherein the water is preferably deionized water, and the volume of the water is preferably 10 times that of the fresh cultured rose-stalk callus cells.
The power of the ultrasonic extraction may be the power of ultrasonic extraction conventional in the art, for example 500W; the time of the ultrasonic extraction may be an extraction time conventional in the art, for example 30 minutes.
The rate of centrifugation may be a centrifugation rate conventional in the art, such as 5000rmp; the centrifugation time may be a centrifugation time conventional in the art, for example 30min.
The pore size of the filter membrane used for the filtration may be conventional in the art, preferably 0.45 μm.
Furthermore, the invention also provides an application of the extract of the callus cells of the rosa tenuifolia in preparing external skin preparations, wherein the extract of the callus cells of the rosa tenuifolia is used as an antioxidant active ingredient, an anti-aging active ingredient or a whitening active ingredient in the external skin preparations.
Among them, the external preparation for skin may be a preparation for external use which is conventional in the art, for example, a whitening, antioxidant or anti-aging cosmetic.
On the basis of conforming to the common knowledge in the field, the above preferred conditions can be arbitrarily combined to obtain the preferred examples of the invention.
The reagents and materials used in the present invention are commercially available.
The invention has the positive progress effects that:
1) According to the invention, vitamin C, nicotinamide, tranexamic acid and hydroxypropyl tetrahydropyran triol are added into the fine-stalk rose callus culture medium, and the synergistic combination of the four components can improve and promote proliferation of fine-stalk rose callus cells, and the content of active ingredients in the fine-stalk rose callus cell extract obtained by culture and the protection of the cell extract on oxidative damage of skin cells are obviously improved, so that the fine-stalk rose callus culture medium has good skin antioxidation effect.
2) Furthermore, the extract of the callus cells of the rosa tenuifolia, which is obtained by culturing the callus culture medium of the rosa tenuis of the invention, can also promote the effect of promoting the proliferation of skin cells (HaCat cells and FB cells), inhibiting tyrosinase and the like, and has good application prospects in the aspects of promoting the regeneration of skin cells, resisting aging, whitening and the like.
Detailed Description
The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.
In the following examples, the sources of the raw materials are as follows:
callus cells of rose tenuifolia:
leaves of a sterile fine-stalk rose (Rosa gracififolia) plantlet were cut to a length of about 0.8mm and inoculated right side up into a culture medium of the formula 1/2MS+6-BA0.2mg/L+α -NAA0.1mg/L+30 g/L of sucrose+6 g/L of agar to induce callus. And taking a proper amount of callus cells as required for subsequent experiments.
The MS medium formulation is shown in Table a below:
table a
| Component (A) | Content (mg/L) |
| NH 4 NO 3 | 1650 |
| KNO 3 | 1900 |
| Anhydrous CaCl 2 | 332.2 |
| Anhydrous MgSO 4 | 180.7 |
| KH 2 PO 4 | 170 |
| KI | 0.83 |
| H 3 BO 3 | 6.2 |
| MnSO 4 ·H 2 O | 16.9 |
| ZnSO 4 ·7H 2 O | 8.6 |
| Na 2 MoO 4 ·2H 2 O | 0.25 |
| CuSO 4 ·5H 2 O | 0.025 |
| CoCl 2 ·6H 2 O | 0.025 |
| FeSO 4 ·7H 2 O | 27.8 |
| Na 2 -EDTA·2H 2 O | 37.26 |
| Inositol (inositol) | 100 |
| Nicotinic acid | 0.5 |
| Pyridoxine hydrochloride (vitamin B) 6 ) | 0.5 |
| Thiamine hydrochloride (vitamin B) 1 ) | 0.1 |
| Glycine (Gly) | 2.0 |
Glacier water is obtained from the middle north slope of Himalayan mountain, and at 5128 m of day karya Bacounty Qu Dengni ma glacier.
In the following examples, the composition of medium A was: MS+2,4-D3.0 mg/L+KT0.5 mg/L+sucrose 30 g/L+agar 7g/L.
In the following examples, the% of the content of hydroxypropyl-tetrahydropyran triol means the mass percentage of the hydroxypropyl-tetrahydropyran triol in the whole medium.
In the following examples and comparative examples, the preparation method of the medium is as follows: taking a reagent MS culture medium, 2, 4-dichlorophenoxyacetic acid (2, 4-D), kinetin (KT), sucrose, agar and glacial water, preparing the culture medium according to a formula MS, 2, 4-D3.0 mg/L, KT 0.5.5 mg/L, 30g/L of sucrose and 7g/L of agar, regulating pH to about 5.8, sterilizing in a sterilizing pot at 121 ℃ for 20min for standby, and adding other components into the culture medium when the temperature of the culture medium is reduced to below 65 ℃.
Example 1
The culture medium comprises the following components: culture medium A, vitamin C10mg/L, nicotinamide 10mg/L, hydroxypropyl tetrahydropyran triol 0.002%, and tranexamic acid 20mg/L.
Example 2
The culture medium comprises the following components: culture medium A, vitamin C1mg/L, nicotinamide 1mg/L, hydroxypropyl tetrahydropyran triol 0.002%, and tranexamic acid 1mg/L.
Example 3
The culture medium comprises the following components: culture medium A, vitamin C20 mg/L, nicotinamide 20mg/L, hydroxypropyl tetrahydropyran triol 0.005% and tranexamic acid 20mg/L.
Example 4
The culture medium comprises the following components: culture medium A, vitamin C50 mg/L, nicotinamide 50mg/L, hydroxypropyl tetrahydropyran triol 0.01% and tranexamic acid 100mg/L.
Examples 5 to 19
Example 5: adding 10mg/L vitamin C to the culture medium A
Example 6: adding 50mg/L vitamin C to the culture medium A
Example 7: adding 100mg/L vitamin C to the culture medium A
Example 8: adding 1000mg/L vitamin C into the culture medium A
Example 9: adding nicotinamide 10mg/L on culture medium A
Example 10: adding nicotinamide 50mg/L on culture medium A
Example 11: adding nicotinamide 100mg/L on culture medium A
Example 12: nicotinamide 1000mg/L was added to medium A
Example 13: adding 1mg/L of tranexamic acid to the culture medium A
Example 14: adding 10mg/L of tranexamic acid to the culture medium A
Example 15: adding 100mg/L of tranexamic acid to the culture medium A
Example 16: adding 1000mg/L of tranexamic acid to the culture medium A
Example 17: adding hydroxypropyl tetrahydropyran triol 0.01% to Medium A
Example 18: adding hydroxypropyl tetrahydropyran triol 0.1% to Medium A
Example 19: adding hydroxypropyl tetrahydropyran triol 1% to culture medium A
Effect example 1 cultivation of callus cells of Rosa tenuifolia and Effect of different culture Medium on proliferation of callus cells of Rosa tenuifolia
Culturing the callus cells of the rose tenuifolia: the culture mediums prepared according to the components of the examples and the comparative examples are adopted, each treatment is carried out in parallel, the volume of the culture medium of each culture bottle is 50mL, about 0.5g of roseus aculeatus callus cells are inoculated in the culture bottle, the culture conditions are that the temperature is 20+/-1 ℃, the humidity is 60% -75%, the illumination intensity is 1500 Lx-2000 Lx, the illumination is carried out for 12 hours every day, and the culture time is 20 days.
As is clear from the results of Table 1, the addition of an active ingredient at an appropriate concentration significantly promoted the proliferation of the fine rose cells, and the proliferation promoting effects of examples 1 to 4, example 6, example 10, example 15 and example 17 were the best. These 8 media were therefore selected for further investigation.
TABLE 1 multiplication factor of fine rose cells cultured in different media
| Sample of | Multiplication factor | Growth state |
| Medium A cells | 4.3 times | Preferably, it is |
| Example 1 cells | 5.3 times | Good (good) |
| Example 2 cells | 3.6 times | Preferably, it is |
| Example 3 cells | 4.5 times | Preferably, it is |
| Example 4 cells | 2.9 times | In general |
| Example 5 cells | 4.1 times | Good (good) |
| Example 6 cells | 6.5 times | Good (good) |
| Examples7-cells | 1.9 times | In general |
| Example 8 cells | 0.2 times | Difference of difference |
| Example 9 cells | 3.7 times | Preferably, it is |
| EXAMPLE 10 cells | 5.3 times | Good (good) |
| Example 11 cells | 1.9 times | In general |
| EXAMPLE 12 cells | 0.3 times of | Difference of difference |
| Example 13 cells | 2.5 times | In general |
| EXAMPLE 14 cells | 3.2 times | Preferably, it is |
| EXAMPLE 15 cells | 4.6 times | Preferably, it is |
| EXAMPLE 16 cells | 2.4 times | In general |
| EXAMPLE 17 cells | 5.5 times | Good (good) |
| EXAMPLE 18 cells | 3.9 times | Preferably, it is |
| EXAMPLE 19 cells | 0.43 times | Difference of difference |
Effect example 2 extracts of fine rose cells obtained from different media
Fresh rose cells (1.5-2.0 g) obtained after the culture in the above effect example 1 were taken, accurately weighed, ground and crushed by a grinder (60 Hz,60 s. Times.2 times), then 10 times of deionized water was added, 500W ultrasonic extraction was performed for 30min,5000rmp centrifugation was performed for 30min, and the supernatant was taken and filtered at 0.45. Mu.M to obtain rose cell extract.
1. Determination of the solid content in the extracts of the cells of Rosa tenuifolia obtained from different combinations of culture media
A certain volume of the extract was dried and weighed to determine the solid content (see table 2). From the results shown in Table 2, it was found that the medium containing the active ingredient can promote the accumulation of substances in the cells of Rosa tenuifolia.
TABLE 2 extraction yields of various combinations of Rosa tenuifolia cell extracts
| Sample of | Solid content (mg/mL) |
| Culture medium A-cell extract | 2.82 |
| Example 1 cell extracts | 3.77 |
| Example 2 cell extracts | 3.26 |
| EXAMPLE 3 cell extracts | 3.98 |
| Example 4 cell extracts | 4.07 |
| EXAMPLE 6 cell extracts | 3.30 |
| EXAMPLE 10 cell extracts | 3.58 |
| EXAMPLE 15 cell extracts | 3.86 |
| EXAMPLE 17 cell extracts | 3.23 |
2. Determination of the polyphenol content in the cell extracts of Rosa tenuifolia obtained from different combinations of culture media
Total polyphenol content FoThe lin-Ciocalteu method uses gallic acid as a reference substance for detection. Taking 0.1mL of gallic acid standard solution and sample solution to be tested, adding 0.5mL of 10% Folin-Ciocalteu reagent for 3 times in parallel, mixing, standing at room temperature for 5min, adding 0.4mL of 7.5% NaCO 3 The solution was left at room temperature for 60 minutes after mixing, and the OD at 765nm was measured. The content of polyphenols in the samples was calculated from the standard curves by plotting the OD520 of each tube of the standard solution against the content of gallic acid (see Table 3).
From the results of Table 3, it is understood that examples 6, 10 and 17 can promote the synthesis of polyphenol substances by the cells of Rosa tenuifolia.
TABLE 3 polyphenol content in Rosa tenuifolia cell extracts
| Sample of | Polyphenol content (%) |
| Culture medium A-cell extract | 3.6％ |
| Example 1 cell extracts | 3.5％ |
| Example 2 cell extracts | 3.4％ |
| Example 3 cell extracts | 3.5％ |
| Example 4 cell extracts | 3.4％ |
| Example 6 cell extracts | 4.1％ |
| Example 10 cell extracts | 4.0％ |
| Example 15 cell extracts | 3.5％ |
| Example 17 cell extracts | 4.0％ |
3. Vc content in rose cell extracts obtained from different combination of culture media
The Vc content of the rose-pelargonium cell extract was determined using LS50B fluorescence detector ((SNF/instrument-106-2001).
TABLE 4 Vc content in Rosa tenuifolia cell extracts
4. Nicotinamide content in Rosa tenuifolia cell extracts obtained from different combinations of culture media
The content of nicotinamide is determined by high performance liquid chromatography, and the determination method is described in Lin Yikan, gu Yuxiang, wang Chengping et al, which refer to high performance liquid chromatography for determining the content of nicotinamide in cosmetics, perfume and essence cosmetics, 2015, 003 (29-32).
TABLE 5 nicotinamide content in Rosa tenuifolia cell extracts
5. Content of tranexamic acid in the rose cell extract obtained from different combinations of culture media
Determination of tranexamic acid content reference Tangping, tang Xin et al, high performance liquid chromatography for determination of tranexamic acid content in cosmetics, daily chemical industry, 2018,1 (57-60).
TABLE 6 content of tranexamic acid in Rosa tenuifolia cell extract
Effect example 3
1. Effect of the extracts of Rosa tenuifolia cells obtained from different combinations of media on the free radical scavenging Rate
1, 1-diphenyl-2-trinitrophenylhydrazine (DPPH) is a stable nitrogen-centered organic radical. DPPH method was proposed in 1958 and is widely used to measure the anti-aging capacity of biological samples, classified substances and foods. The method is based on the fact that DPPH free radical has single electron, has strong absorption at 517nm, and the alcohol solution is purple. When the free radical scavenger exists, the free radical scavenger is paired with single electron to make its absorption gradually disappear, and its fading degree is quantitatively related to the number of electrons accepted by the free radical scavenger, so that a spectrophotometer can be used for quick quantitative analysis to detect the free radical scavenger, thereby evaluating the anti-aging capability of the sample.
Adding 0.1mL of extract sample solution into a test tube, adding 0.1mL of DPPH ethanol solution, mixing, reacting at room temperature in dark place for 30min at room temperature in dark place, measuring OD value at 525nm, and calculating clearance according to the following formula: clearance I (%) = [1- (T-T0)/(C-C0) ] ×100%, wherein: t0: absorbance of 0.1mL sample solution plus 0.1mL 95% ethanol; t: absorbance of 0.1mL sample solution plus 0.1mL ldpph solution; c0: absorbance of 0.1mL water plus 0.1mL 95% ethanol; c: absorbance of 0.1mL water plus 0.1mL DPPH solution.
As is clear from the results in Table 7, the DPPH clearance of the cell extracts of Rosa tenuifolia in examples 1 to 4 and examples 6, 10, 15 and 17 was lower than that of the control group.
TABLE 7 DPPH clearance of different cell extracts of Rosa tenuifolia
2. Effect of the extract of Rosa tenuifolia cells obtained from different combinations of culture media on skin cell proliferation
Keratinocytes are the most important cells in the epidermis and participate in the formation of the physical barrier of the skin, preventing invasion of adverse factors such as physical, chemical and microbial factors, and protecting the skin. One of the main characteristics of skin after aging is the thinning of the epidermis and the slow rate of wound healing, which is mainly caused by the reduced regenerative capacity of keratinocytes in the epidermis layer and the reduced proliferative capacity of the keratinocyte system. Human skin fibroblasts are the most important cells in the dermis reticulation layer of the skin, and are one of the main repair cells after skin aging and cell damage. It not only can promote migration, proliferation and differentiation of epidermal cells, but also can secrete a large amount of collagen, elastin and various cell repair factors, and has strong self-renewal capacity, so that aged skin can be repaired.
The prepared samples are prepared into a solution by deionized water, added into human immortalized epidermal HaCat cells or human primary Fibroblast (FB) culture solution, and the solvent without the samples is taken as a blank control. After 48 hours of culture, the cells were stained by MTT method, absorbance at 550nm was measured by an enzyme-labeled instrument, the proliferation rate of the blank control was 100%, and the effect on HaCat or FB cell proliferation was evaluated by reference to the blank control.
As is clear from the results shown in Table 8, the extract of fine rose cells obtained by culturing the following combination medium has a certain proliferation promoting effect on both skin cell epidermis HaCat cells and fibroblasts, and the promoting effect of examples 1 and 3 and examples 10, 15 and 17 is stronger than that of the medium A group, and the promoting effect of example 1 is the best.
TABLE 8 Effect of different combinations of Rosa tenuifolia cell extracts on skin cell proliferation
3. Protective effect of Rosa tenuifolia cell extracts obtained from different combinations of culture media on oxidative damage of skin cells
The free radicals can cause various irreversible oxidative damages to organisms at the cellular level, the molecular level and even the tissue organ level, accelerate the aging process of organism cells and even the whole organism, and induce various diseases related to aging. Hydrogen peroxide (hydrogen peroxide, H) 2 O 2 ) Is an important oxygen free radical in cells, and can cause direct oxidative stress reaction and oxidative damage caused by oxidative stress to human skin cells. In vitro utilization of H 2 O 2 Can induce and accelerate aging process of cells caused by oxidative stress, and simulate pathological process of oxidative damage in vivo.
Human immortalized epidermic HaCat cells or human primary Fibroblasts (FB) are planted in 96-well plates, cultured for 24 hours, and then added with drugs and H 2 O 2 (1 mM) culture medium for 24 hours, and MTT method was used to measure cell viability. By not adding H 2 O 2 Treated as a control group, the cell viability of the control was 100%, the control was referenced to the control, and H was evaluated 2 O 2 And the effect of sample processing on cell viability.
From the results shown in Table 9, the cell extracts of examples 1 to 4 have an obvious protective effect on the cell oxidative damage caused by hydrogen peroxide, and the cell extracts of examples have an obvious protective effect on the cell oxidative damage caused by hydrogen peroxide in the HaCaT cell model, and the protective effect is stronger than that of the culture medium A group. Among the two cell oxidative damage models, the protection effect of the combination of the four effective components is best. In combination with the DPPH clearance test result, the clearance rate of the control group is higher than that of the test group, and it is speculated that the extract of the rosa tenuifolia cells cultured by adding other functional components into the culture medium may have different antioxidant pathways compared with the culture medium A.
TABLE 9 protection of skin oxidative damage by different combinations of Rosa tenuifolia cell extracts
4. Ultraviolet injury protection effect of Rosa tenuifolia cell extracts obtained from different combination culture mediums
Aging of skin is classified into endogenous aging and exogenous aging, and sunlight, particularly Ultraviolet (UV) irradiation, is a major factor in the formation of exogenous aging, so exogenous aging is also called photoaging. Associated with skin photoaging are mainly UVA (320 nm-400 nm) and small amounts of UVB (280-320 nm). Many data indicate that 95% of the uv light that is irradiated to the skin is absorbed by keratinocytes, and that exposure of the skin to UVA radiation generates ROS and causes various damage, including oxidative stress. Ultraviolet UVB directly acts on epidermal cells, and the skin barrier function is further affected by the induction of oxidative stress and inflammatory processes, so that photodamage is formed. We therefore established a model of photodamage to skin by irradiation of dermal Fibroblast fibroblastist cells with UVA/UVB, and used this model to evaluate the protective effect of actives on skin uv damage.
Human immortalized epidermal HaCat cells or human primary Fibroblasts (FB), seeded in 96-well plates at 37 ℃ with 5% CO 2 Culturing in a cell culture box for 24 hours. Then adding the mixture into the whole culture medium containing the drug or solvent control for culturing for 24 hours. The medium was replaced with PBS and subjected to UVA/UVB irradiation at a dose of 10J/cm 2 Or 40J/cm 2 . After PBS was removed, the culture medium containing the sample to be tested was added and the culture was continued for 24 hours, and the MTT method was used to measure the cell viability. The non-UVA/UVB-irradiated group was used as a blank control group (UVA-/UVB-), and the UVA/UVB-irradiated group was used as a control group (UVA+/UVB+). Cell viability of the blank was100%, reference blank, assess the effect of UVA/UVB and sample treatment on cell viability.
As is clear from the results in Table 10-1, the cell extracts of Rosa tenuifolia obtained from the different combinations of the following media had less cell damage to UVA than the group A media, and had no significant protective effect on UVA.
TABLE 10-1 protection of skin UVA injury by Rosa tenuifolia cell extract
As is clear from the results in Table 10-2, the cell extracts of Rosa tenuifolia had a remarkable protective effect against UVB-induced cell damage, and the cell extracts of examples 6 and 10 were remarkably more protective than the group A medium.
TABLE 10-2 protection of skin UVB lesions by different Rosa tenuifolia cells
5. Whitening effect of rose cell extracts obtained from different combination of culture media
Recent studies have demonstrated that pigmentation is caused by ultraviolet light activating melanogenesis enzymes in melanocytes in the epidermis, producing pigment. Melanocytes in basal lamina in the human epidermis are in contact with and associated with surrounding keratinocytes to form "epidermal melanocytes," where melanin is a nitrogen-containing complex synthesized in melanosomes. Generally, the amount and quality of melanin in the skin is an important factor in determining the skin color. In the melanin production process, tyrosinase converts tyrosine into dopa, and dopaquinone is polymerized into macromolecular melanin through non-enzymatic oxidation. Thus, by inhibiting tyrosinase activity, melanin production can be restricted and skin pigmentation can be improved.
The skin color is derived from melanin stored within keratinocytes. Generally, people with increased storage melanin are darker in their skin and are also more protected from solar radiation. The amount and quality of melanin is an important factor in determining skin color in skin melanin. Tyrosinase is a copper-containing oxidoreductase with a complex structure, and is widely used in microorganisms, animals, plants and human bodies. Tyrosinase is a key enzyme in the synthesis of melanin by the skin in humans.
The inhibition of tyrosinase by the samples was determined by L-Dopa oxidation. The melanoma B16 cells of the mice are planted in a 96-well plate, after 24 hours, the complete culture medium containing the drugs is added for culturing for 48 hours, the complete culture medium is washed by PBS for 2 times, 1 percent TritonX-100 (in PBS) is added, and the mixture is quickly placed in a refrigerator at-80 ℃ for 30 minutes. Thawing at room temperature or 37 ℃, lysing cells, and shaking and mixing uniformly. Adding 0.2% dopa water solution, shaking and mixing, reacting at 37 ℃ for 3 hours, and measuring OD490nm. The enzyme activity was calculated as follows: tyrosinase inhibition ratio = [1- (experimental group OD value/control group OD value ] ×100% ], a group without extract addition was used as a blank control.
From the results shown in Table 11, the extract of the cells of Rosa tenuifolia of example 1 and example 17 has an inhibitory effect on the tyrosinase activity of the B16 cells of melanoma in mice, and the inhibitory effect is stronger than that of the group A culture medium.
TABLE 11 Effect of different Rosa tenuifolia cell extracts on the tyrosinase activity of murine melanoma B16 cells
| Sample of | Testing concentration (mg/mL) | Tyrosinase inhibition rate (%) |
| Blank control group | - | 0 |
| Culture medium A-cell extract | 0.5 | 20 |
| Example 1 cell extracts | 0.5 | 28 |
| Example 2 cell extracts | 0.5 | 21 |
| EXAMPLE 3 cell extracts | 0.5 | 25 |
| Example 4 cell extracts | 0.5 | 22 |
| EXAMPLE 6 cell extracts | 0.5 | 10 |
| Example 10 cell extracts | 0.5 | 17 |
| Example 15 cell extracts | 0.5 | 22 |
| Example 17 cell extracts | 0.5 | 29 |
Taken together, the results show that, compared with the culture medium A, examples 1-4 are significantly improved in terms of resisting oxidative damage to both HaCaT skin cells and FB skin cells, while examples 5-19 are not improved in terms of resisting oxidative damage to both HaCaT and FB skin cells. The example 1 is superior to the culture medium A in promoting the proliferation of the fine-stalk rose cells, and the fine-stalk rose extract has the effects of promoting the proliferation of HaCaT and FB skin cells, resisting oxidation injury, inhibiting tyrosinase activity and the like, and has a slightly improved UVA resistance to the HaCaT cells.
Claims (26)
1. The callus proliferation culture medium for rosa tenuifolia is characterized by comprising the following components:
MS culture medium;
a phytohormone, said phytohormone being 2, 4-D3 mg/L and KT 0.5 mg/L;
The active ingredients comprise 5-45 mg/L of vitamin C, 5-45 mg/L of nicotinamide, 10-50 mg/L of tranexamic acid and 0.002-0.01% of hydroxypropyl tetrahydropyran triol;
wherein: mg/L refers to the concentration in the callus proliferation medium of the fine rose;
the percentage refers to the mass percentage of the hydroxypropyl tetrahydropyran triol in the callus proliferation culture medium of the rosa tenuifolia.
2. The callus proliferation medium of fine rose according to claim 1, wherein the concentration of vitamin C in the callus proliferation medium of fine rose is 5-40 mg/L;
and/or the concentration of the nicotinamide in the callus proliferation medium of the rose sessilifolia is 5-40 mg/L;
and/or the concentration of the tranexamic acid in the callus proliferation medium of the rose sessilifolia is 10-40 mg/L;
and/or, the hydroxypropyl tetrahydropyran triol accounts for 0.002-0.009% of the callus proliferation culture medium of the rosa tenuifolia;
and/or the mass ratio of the vitamin C to the nicotinamide to the tranexamic acid is 1:1:2;
and/or, the callus proliferation culture medium of the rosa tenuifolia also comprises glacial water, and the water source of the glacial water is Himalayan mountain;
And/or the pH of the callus proliferation culture medium of the rose sessiliflora is 5-7;
and/or, the rose-stalk callus proliferation medium further comprises a carbon source;
and/or, the callus proliferation medium of the rosa tenuifolia further comprises a coagulating agent.
3. The roseus tenuifolia callus proliferation medium according to claim 2, wherein the concentration of vitamin C in the roseus tenuifolia callus proliferation medium is 10-20mg/L;
and/or the concentration of the nicotinamide in the callus proliferation medium of the fine-stalk rosa rugosa is 10-20mg/L;
and/or the concentration of the tranexamic acid in the callus proliferation medium of the fine rose is 18-40 mg/L;
and/or the hydroxypropyl tetrahydropyran triol accounts for 0.002-0.005% of the mass of the callus proliferation culture medium of the rosa tenuifolia;
and/or the water source area of glacier water is a north slope in the middle section of a Himalayan mountain;
and/or, the pH of the callus proliferation medium of the rosa tenuifolia is 5.8;
and/or the carbon source is one or more of sucrose, maltose and glucose;
and/or the concentration of the carbon source in the callus proliferation medium of the rosa tenuifolia is 28-35 g/L;
And/or, the coagulant is agar and/or gelatin;
and/or the concentration of the coagulant in the callus proliferation medium of the rose-radicle is 6-8 g/L.
4. The roseus tenuius callus proliferation medium of claim 2, wherein the concentration of vitamin C in the roseus tenuius callus proliferation medium is 5, 8, 10, 12, 14, 15, 20, 25, 30, 35 or 40mg/L;
and/or the concentration of nicotinamide in the callus proliferation medium of rose-capillipes is 5, 8, 10, 15, 20, 25, 30, 35 or 40mg/L;
the concentration of the tranexamic acid in the callus proliferation culture medium of the rose sessiliflorus is 10, 20, 30 and 40;
and/or, the hydroxypropyl tetrahydropyran triol accounts for 0.003%, 0.005%, 0.006%, 0.007%, 0.008% or 0.009% of the mass of the callus proliferation medium of the rosa tenuifolia;
and/or the water intake of glacier water is day karst Bacounty Qu Dengni MABingchuan;
and/or, the carbon source is sucrose;
and/or the concentration of the carbon source in the callus proliferation medium of the fine rose is 30 g/L;
and/or, the coagulant is agar;
and/or the concentration of the coagulant in the callus proliferation medium of the fine rose is 7 g/L.
5. The callus proliferation medium of fine rose according to claim 2, wherein the concentration of vitamin C in the callus proliferation medium of fine rose is 8-12 mg/L;
and/or the concentration of the nicotinamide in the callus proliferation medium of the rose sessiliflora is 8-12 mg/L;
and/or the concentration of the tranexamic acid in the callus proliferation medium of the fine rose is 18-22 mg/L;
and/or the hydroxypropyl tetrahydropyran triol accounts for 0.002-0.003% of the mass of the callus proliferation culture medium of the rosa tenuifolia.
6. The roseus tenuifolia callus proliferation medium according to claim 2, wherein the concentration of vitamin C in the roseus tenuifolia callus proliferation medium is 10 mg/L;
and/or the concentration of the nicotinamide in the callus proliferation medium of the fine-stalk rosa rugosa is 10 mg/L;
and/or the concentration of the tranexamic acid in the callus proliferation medium of the fine rose is 20 mg/L;
and/or, the hydroxypropyl tetrahydropyran triol accounts for 0.002% of the mass of the callus proliferation medium of the rosa tenuifolia.
7. The callus proliferation medium of fine rose according to claim 2, wherein the mass concentration of the vitamin C, the nicotinamide, the tranexamic acid and the hydroxypropyl tetrahydropyran triol in the callus proliferation medium of fine rose is 10 mg/L, 10 mg/L, 20 mg/L and 0.002-0.1% respectively;
Alternatively, the callus proliferation medium of the rosa tenuifolia comprises the following components: MS, 2, 4-D3.0 mg/L, KT 0.5.5 mg/L, sucrose 30 g/L, agar 7 g/L, glacial water, vitamin C10 mg/L, nicotinamide 10mg/L, tranexamic acid 20 mg/L, hydroxypropyl tetrahydropyran triol 0.002%;
or, the callus proliferation culture medium of the rosa tenuifolia consists of the following components: MS, 2, 4-D3.0 mg/L, KT 0.5.5 mg/L, sucrose 30 g/L, agar 7 g/L, glacial water, vitamin C10 mg/L, nicotinamide 10mg/L, tranexamic acid 20 mg/L, hydroxypropyl tetrahydropyran triol 0.002%.
8. The callus proliferation culture medium for rosa tenuifolia is characterized by comprising the following components: MS culture medium;
a phytohormone, said phytohormone being 2, 4-D3 mg/L and KT0.5 mg/L;
the active ingredient is 90-110 mg/L of tranexamic acid.
9. The callus proliferation medium according to claim 8, wherein the concentration of tranexamic acid is 100 mg/L.
10. The fine rose callus proliferation medium according to claim 8, wherein the fine rose callus proliferation medium consists of: MS culture medium, glacier water, 2, 4-D3 mg/L, KT, 0.5 mg/L, sucrose 30 g/L, agar 7 g/L and tranexamic acid 90-110 mg/L.
11. The roseus tenuifolia callus proliferation medium according to claim 10, wherein the concentration of tranexamic acid in the roseus tenuifolia callus proliferation medium is 100 mg/L.
12. The callus proliferation culture medium for rosa tenuifolia is characterized by comprising the following components: MS culture medium;
a phytohormone, said phytohormone being 2, 4-D3 mg/L and KT 0.5 mg/L;
and the active ingredient is hydroxypropyl tetrahydropyran triol accounting for 0.009-0.015%.
13. The roselle callus proliferation medium according to claim 12 wherein the concentration of hydroxypropyl tetrahydropyran triol is 0.01%.
14. The roselle callus proliferation medium of claim 12, wherein the roselle callus proliferation medium consists of: MS culture medium, glacial water, 2, 4-D3 mg/L, KT, 0.5 mg/L, sucrose 30 g/L, agar 7 g/L and hydroxypropyl tetrahydropyran triol 0.009-0.015%.
15. The roselle callus proliferation medium according to claim 14 wherein the concentration of hydroxypropyl tetrahydropyran triol is 0.01%.
16. The preparation method of the callus proliferation culture medium of the rosa tenuifolia is characterized by comprising the following steps of: mixing the components in the callus proliferation medium of fine rose according to any one of claims 1 to 15.
17. The method for preparing a callus proliferation medium of rose-capitula according to claim 16, wherein the method for preparing a callus proliferation medium of rose-capitula further comprises a sterilization step.
18. The method for preparing a callus proliferation medium according to claim 17, wherein the sterilization step is high-temperature sterilization.
19. The method for preparing a callus proliferation medium according to claim 18, wherein when the callus proliferation medium comprises vitamin C, the vitamin C is added after the high temperature sterilization and after the temperature is reduced to 65 ℃ or lower;
or when the callus proliferation medium of the rosa tenuifolia comprises nicotinamide, after the high-temperature sterilization, the nicotinamide is added after the temperature is reduced to below 65 ℃;
or when the callus proliferation culture medium of the rosa tenuifolia comprises tranexamic acid, after the high-temperature sterilization, the tranexamic acid is added after the temperature is reduced to below 65 ℃;
or when the callus proliferation medium of the rosa tenuifolia comprises hydroxypropyl tetrahydropyran triol, after the high-temperature sterilization, adding the hydroxypropyl tetrahydropyran triol after the temperature is reduced to below 65 ℃;
Or, when the callus proliferation medium of rosa tenuifolia comprises vitamin C, nicotinamide, tranexamic acid and hydroxypropyl tetrahydropyran triol, after the high temperature sterilization, the vitamin C, the nicotinamide, the tranexamic acid and the hydroxypropyl tetrahydropyran triol are added after the temperature is reduced to below 65 ℃.
20. An extract of rose-stem callus cells, which is obtained by extracting rose-stem callus cells after culturing, wherein the rose-stem callus cells after culturing are obtained by culturing rose-stem callus with the rose-stem callus proliferation medium according to any one of claims 1 to 15.
21. The roselle callus cell extract of claim 20, wherein the culturing conditions are: the temperature is 20+/-1 ℃, the humidity is 60% -75%, the illumination intensity is 1500 Lx% -2000 Lx, and the illumination is carried out for 12 hours each day;
and/or, the time of the culturing is 20 days;
and/or the solid content of the fine-stalk rosa calluses cell extract is 3-5 mg/mL, wherein the solid content of the fine-stalk rosa calluses cell extract is the mass percentage of the mass of the rest part of the fine-stalk rosa calluses cell extract after drying;
And/or the content of polyphenol in the extract of the calluses of the fine roses is 3.0-4.5%, wherein the content of the polyphenol is the mass percentage of the polyphenol in the extract of the calluses of the fine roses;
and/or the content of Vc in the extract of the calluses of the fine roses is 1-2 mg/kg, and the content of Vc is the mass of Vc in the extract of the calluses of the fine roses per 1kg based on the wet weight of cells;
and/or the content of nicotinamide in the extract of the calluses of the fine roses is 100-600 mg/kg, and the content of nicotinamide is the mass of nicotinamide in the extract of the calluses of the fine roses per 1kg by cell wet weight;
and/or the content of the tranexamic acid in the extract of the calluses of the fine roses is 100-500 mg/kg, and the content of the tranexamic acid is the mass of the tranexamic acid in the extract of the calluses of the fine roses per 1kg by cell wet weight;
and/or the extraction method is ultrasonic extraction.
22. The roselle callus cell extract of claim 21, wherein the roselle callus cell extract has a solids content of 3.0-4.5 mg/mL;
And/or the content of polyphenol in the extract of the calluses of the rosa tenuifolia is 3.4, 3.5, 4.0 or 4.1%;
and/or the Vc content in the extract of the callus cells of the rosa tenuifolia is 1.1, 1.2, 1.5 or 1.7 mg/kg;
and/or, the nicotinamide content of the extract of the callus cells of the rosa tenuifolia is 117.6, 200.8 or 518.6 mg/kg;
and/or the content of tranexamic acid in the extract of the callus cells of the rosa tenuifolia is 120, 216 or 518 mg/kg;
and/or, the ultrasonic extraction comprises the following steps: taking fresh cultured fine-stalk rosa calluses, grinding, adding a proper amount of water, performing ultrasonic treatment, centrifuging, taking supernatant, and filtering to obtain the fine-stalk rosa calluses extract.
23. The roselle callus cell extract of claim 22, wherein the roselle callus cell extract has a solids content of 3.26, 3.77, 3.98 or 4.07 mg/mL;
and/or the grinding speed is 50-70 Hz;
and/or the grinding time is 50-70 s;
and/or the grinding times are 1-3 times;
and/or the water is deionized water, and the volume of the water is 10 times that of the fresh cultured rose-stalk callus cells;
And/or the power of the ultrasonic extraction is 500W;
and/or, the ultrasonic extraction time is 30min;
and/or, the rate of centrifugation is 5000 rpm;
and/or, the centrifugation time is 30min;
and/or the pore size of the filtration membrane used in the filtration is 0.45 μm.
24. The roselle callus cell extract of claim 23, wherein the rate of milling is 60Hz;
and/or, the grinding time is 60s;
and/or the number of grinding is 2.
25. The use of a rose-stalk callus cell extract according to any one of claims 20 to 24 as an antioxidant active ingredient, an anti-aging active ingredient or a whitening active ingredient in a skin external preparation.
26. The use according to claim 25, wherein the external preparation for skin is a whitening, antioxidant or anti-aging cosmetic.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011065238.4A CN114303944B (en) | 2020-09-30 | 2020-09-30 | Callus culture medium and extract of rosa tenuifolia, preparation method and application |
| CN202310835582.4A CN116918701A (en) | 2020-09-30 | 2020-09-30 | Callus culture medium and extract of rosa tenuifolia, preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011065238.4A CN114303944B (en) | 2020-09-30 | 2020-09-30 | Callus culture medium and extract of rosa tenuifolia, preparation method and application |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310835582.4A Division CN116918701A (en) | 2020-09-30 | 2020-09-30 | Callus culture medium and extract of rosa tenuifolia, preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114303944A CN114303944A (en) | 2022-04-12 |
| CN114303944B true CN114303944B (en) | 2023-08-11 |
Family
ID=81032259
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310835582.4A Pending CN116918701A (en) | 2020-09-30 | 2020-09-30 | Callus culture medium and extract of rosa tenuifolia, preparation method and application |
| CN202011065238.4A Active CN114303944B (en) | 2020-09-30 | 2020-09-30 | Callus culture medium and extract of rosa tenuifolia, preparation method and application |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310835582.4A Pending CN116918701A (en) | 2020-09-30 | 2020-09-30 | Callus culture medium and extract of rosa tenuifolia, preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN116918701A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116918701A (en) * | 2020-09-30 | 2023-10-24 | 伽蓝（集团）股份有限公司 | Callus culture medium and extract of rosa tenuifolia, preparation method and application |
Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007107803A2 (en) * | 2006-03-23 | 2007-09-27 | Adrienne Seres | Process and modified media for preparing callus- and cell suspension cultures of hypericum perforatum l. |
| CN101139571A (en) * | 2007-08-29 | 2008-03-12 | 清华大学 | Method for producing saussurea involucrate secondary metabolites |
| CN106236674A (en) * | 2016-09-22 | 2016-12-21 | 上海新高姿化妆品有限公司 | A kind of compositions with white-skinned face function and the application in cosmetics thereof |
| JP2017132694A (en) * | 2016-01-25 | 2017-08-03 | 日本メナード化粧品株式会社 | Methods for preparing callus of epiphyllum oxypetalum and external and internal preparations for skin containing the extract of the callus as effective ingredients |
| CN108419671A (en) * | 2018-01-19 | 2018-08-21 | 安徽霍山县松鹤堂中药材有限公司 | A kind of stem tip tissue culture breeding and seedling method of David's-harp |
| CN109646381A (en) * | 2019-03-05 | 2019-04-19 | 陈伟伟 | A kind of moisturizing lotion and preparation method thereof added with plant extracts |
| CN109880789A (en) * | 2019-04-25 | 2019-06-14 | 杨友霞 | A kind of print chinaberry cell suspension cultures base and cell suspension cultures method |
| CN110724659A (en) * | 2019-09-27 | 2020-01-24 | 伽蓝（集团）股份有限公司 | Anti-browning culture method for jade dew cells |
| CN111202777A (en) * | 2020-03-06 | 2020-05-29 | 伽蓝（集团）股份有限公司 | A Rosa tenuifolia callus cell extract and its application in preparing skin external preparation |
| CN111280063A (en) * | 2020-04-02 | 2020-06-16 | 安赛搏(重庆)生物技术有限公司 | Rapid callus induction and large-scale suspension culture method for desert rose |
| WO2020188535A1 (en) * | 2019-03-21 | 2020-09-24 | Demethra Biotech S.R.L. | Phytocomplex and extract of a meristematic cell line selected from echinacea purpurea |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1675992A (en) * | 2005-04-19 | 2005-10-05 | 扬州大学 | Chinese herbeceous peony invitro tissue culture method |
| JP3830960B1 (en) * | 2005-08-12 | 2006-10-11 | 建仁 河乃 | Hair growth substance |
| CN101486995B (en) * | 2009-02-24 | 2011-01-05 | 中国检验检疫科学研究院 | Culture medium for tissue engineering skin convey and formulating method thereof |
| CN102440192B (en) * | 2011-10-14 | 2013-04-10 | 扬州大学 | Culture media for peony high frequency embryogenic callus differentiation, and culture method thereof |
| KR101791641B1 (en) * | 2016-03-30 | 2017-10-30 | (주)셀트리온 | Nelumbo Nucifera callus extract with an enhanced content of gallic acid, manufacturing method of the same and composition for skin whitening comprising the same |
| CN116918701A (en) * | 2020-09-30 | 2023-10-24 | 伽蓝（集团）股份有限公司 | Callus culture medium and extract of rosa tenuifolia, preparation method and application |
-
2020
- 2020-09-30 CN CN202310835582.4A patent/CN116918701A/en active Pending
- 2020-09-30 CN CN202011065238.4A patent/CN114303944B/en active Active
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007107803A2 (en) * | 2006-03-23 | 2007-09-27 | Adrienne Seres | Process and modified media for preparing callus- and cell suspension cultures of hypericum perforatum l. |
| CN101139571A (en) * | 2007-08-29 | 2008-03-12 | 清华大学 | Method for producing saussurea involucrate secondary metabolites |
| JP2017132694A (en) * | 2016-01-25 | 2017-08-03 | 日本メナード化粧品株式会社 | Methods for preparing callus of epiphyllum oxypetalum and external and internal preparations for skin containing the extract of the callus as effective ingredients |
| CN106236674A (en) * | 2016-09-22 | 2016-12-21 | 上海新高姿化妆品有限公司 | A kind of compositions with white-skinned face function and the application in cosmetics thereof |
| CN108419671A (en) * | 2018-01-19 | 2018-08-21 | 安徽霍山县松鹤堂中药材有限公司 | A kind of stem tip tissue culture breeding and seedling method of David's-harp |
| CN109646381A (en) * | 2019-03-05 | 2019-04-19 | 陈伟伟 | A kind of moisturizing lotion and preparation method thereof added with plant extracts |
| WO2020188535A1 (en) * | 2019-03-21 | 2020-09-24 | Demethra Biotech S.R.L. | Phytocomplex and extract of a meristematic cell line selected from echinacea purpurea |
| CN109880789A (en) * | 2019-04-25 | 2019-06-14 | 杨友霞 | A kind of print chinaberry cell suspension cultures base and cell suspension cultures method |
| CN110724659A (en) * | 2019-09-27 | 2020-01-24 | 伽蓝（集团）股份有限公司 | Anti-browning culture method for jade dew cells |
| CN111202777A (en) * | 2020-03-06 | 2020-05-29 | 伽蓝（集团）股份有限公司 | A Rosa tenuifolia callus cell extract and its application in preparing skin external preparation |
| CN111280063A (en) * | 2020-04-02 | 2020-06-16 | 安赛搏(重庆)生物技术有限公司 | Rapid callus induction and large-scale suspension culture method for desert rose |
Non-Patent Citations (1)
| Title |
|---|
| 李惠玲等.细梗蔷薇愈伤组织提取物在人真皮成纤维细胞及3D表皮模型上的护肤功效探究.日用化学工业（中英文）.2023,第53卷(第03期),第300-307页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114303944A (en) | 2022-04-12 |
| CN116918701A (en) | 2023-10-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106038386B (en) | Composition with white-skinned face function and preparation method and application | |
| CN111096941B (en) | Whitening composition containing symbiotic bacteria combined fermentation product, whitening essence and preparation method of whitening essence | |
| KR20080001791A (en) | Skin anti-wrinkle cosmetics composition containing forsythiae fruit extract | |
| KR101824770B1 (en) | Anti-wrinkle cosmetic composition comprising essentially Polygonum multiflorum adventitious extract | |
| CN114303944B (en) | Callus culture medium and extract of rosa tenuifolia, preparation method and application | |
| CN114533618A (en) | Whitening and repairing composition and preparation method and application thereof | |
| KR100521768B1 (en) | Cosmetic Compositions for a Skin Whitening | |
| CN111202777B (en) | A Rosa tenuifolia callus cell extract and its application in preparing skin external preparation | |
| KR20110018091A (en) | Composition for skin whitening | |
| CN112386547B (en) | Skin health care application of pineapple extract | |
| KR20210144725A (en) | Anti-aging, antioxidant, anti-inflammatory and whitening agents, and cosmetics | |
| KR101995557B1 (en) | Cosmetic composition including fermented water lily extract and method for manufacturing the same | |
| KR20180039301A (en) | Oriental cosmetic composition comprising the extract of coptis chinensis | |
| KR20200098153A (en) | Composition for preventing or improving of skin wrinkle or skin photo-aging comprising fermented extract of Dendropanax morbifera and purple sweet potato as an active ingredient | |
| CN113384510B (en) | Fermented composition with anti-photoaging, relieving and cooling effects and preparation method and application thereof | |
| KR20120092229A (en) | Cosmetic composition comprising the fermented extracts of tissue cultures of brassica napus l, punica granatum and panax ginseng) | |
| KR20090081956A (en) | Skin whitening cosmetics composition containing an effective ingredient extracted from Acanthopanax sessiliflorum Seeman and manufacturing method thereof | |
| KR102689835B1 (en) | Cosmetic composition comprising upland rice fermented extracts | |
| KR100837517B1 (en) | Peptide for suppressing melanogenesis, preparation method, and skin external application composition comprising the said peptide | |
| KR20020035925A (en) | Cosmetics composition comprising extract powder with vitis vinifera l.fermented solution and having whitening and anti-wrinkle effect | |
| KR20180052151A (en) | Anti-aging and anti-wrinkle of skin improving cosmetic composition comprising the extract of Sparassis crispa as an active ingredient and preparation method of the same | |
| KR20100073275A (en) | Cosmetics comprising cistanche tubulosa extract and method of making the same | |
| CN114437242B (en) | Usnic polysaccharide, preparation method and application thereof | |
| KR20150081201A (en) | Cosmetic composition for whitening and manufacturing method thereof | |
| KR101810825B1 (en) | Cosmetics compositions containing fermentation extract from leaf Daphniphyllum macropodum Miq. |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CP03 | Change of name, title or address |
Address after: 201403 Shanghai City, Fengxian District Lifenglu No. 12 Patentee after: Shanghai Natural Hall Group Co.,Ltd. Address before: 201403 Shanghai City, Fengxian District Lifenglu No. 12 Patentee before: JALA Group Co. |
|
| CP03 | Change of name, title or address |