CN113694201B - 用于控制生物体内热量产生的组合物、方法及用途 - Google Patents
用于控制生物体内热量产生的组合物、方法及用途 Download PDFInfo
- Publication number
- CN113694201B CN113694201B CN202110986914.XA CN202110986914A CN113694201B CN 113694201 B CN113694201 B CN 113694201B CN 202110986914 A CN202110986914 A CN 202110986914A CN 113694201 B CN113694201 B CN 113694201B
- Authority
- CN
- China
- Prior art keywords
- binding
- ebi
- receptor
- parameter
- organism
- 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
- 230000020169 heat generation Effects 0.000 title claims abstract description 37
- 238000000034 method Methods 0.000 title claims abstract description 35
- 239000000203 mixture Substances 0.000 title claims abstract description 16
- 102100036678 Interleukin-27 subunit alpha Human genes 0.000 claims abstract description 99
- 108010066979 Interleukin-27 Proteins 0.000 claims abstract description 64
- 230000036760 body temperature Effects 0.000 claims abstract description 5
- 150000001875 compounds Chemical class 0.000 claims description 36
- 102100040066 Interleukin-27 receptor subunit alpha Human genes 0.000 claims description 34
- 108040010246 interleukin-27 receptor activity proteins Proteins 0.000 claims description 34
- 238000004519 manufacturing process Methods 0.000 claims description 29
- 238000012360 testing method Methods 0.000 claims description 27
- 238000005259 measurement Methods 0.000 claims description 18
- 230000005764 inhibitory process Effects 0.000 claims description 11
- 238000012216 screening Methods 0.000 claims description 11
- 230000001737 promoting effect Effects 0.000 claims description 8
- 230000009467 reduction Effects 0.000 claims description 7
- 239000012634 fragment Substances 0.000 claims description 6
- 230000003247 decreasing effect Effects 0.000 claims description 4
- 241000124008 Mammalia Species 0.000 claims description 3
- 238000010171 animal model Methods 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 230000002401 inhibitory effect Effects 0.000 claims description 2
- 244000005700 microbiome Species 0.000 claims description 2
- 230000001413 cellular effect Effects 0.000 claims 1
- 238000002560 therapeutic procedure Methods 0.000 claims 1
- 241000699670 Mus sp. Species 0.000 abstract description 74
- 210000001789 adipocyte Anatomy 0.000 abstract description 26
- 239000000539 dimer Substances 0.000 abstract description 21
- 230000001276 controlling effect Effects 0.000 abstract description 19
- 108090000623 proteins and genes Proteins 0.000 abstract description 15
- 230000007783 downstream signaling Effects 0.000 abstract description 14
- 102000004169 proteins and genes Human genes 0.000 abstract description 11
- 102100029820 Mitochondrial brown fat uncoupling protein 1 Human genes 0.000 abstract description 4
- 108050002686 Mitochondrial brown fat uncoupling protein 1 Proteins 0.000 abstract description 4
- 230000037149 energy metabolism Effects 0.000 abstract description 4
- 230000001105 regulatory effect Effects 0.000 abstract description 4
- 230000002631 hypothermal effect Effects 0.000 abstract description 3
- 230000009456 molecular mechanism Effects 0.000 abstract description 3
- 230000008569 process Effects 0.000 abstract description 3
- 238000011160 research Methods 0.000 abstract description 3
- 230000001225 therapeutic effect Effects 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 32
- 108010050258 Mitochondrial Uncoupling Proteins Proteins 0.000 description 24
- 102000015494 Mitochondrial Uncoupling Proteins Human genes 0.000 description 24
- 101100340196 Mus musculus Il27ra gene Proteins 0.000 description 22
- 238000004458 analytical method Methods 0.000 description 22
- 102000005962 receptors Human genes 0.000 description 21
- 108020003175 receptors Proteins 0.000 description 21
- 230000000638 stimulation Effects 0.000 description 21
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 19
- 238000002474 experimental method Methods 0.000 description 18
- 210000003486 adipose tissue brown Anatomy 0.000 description 17
- 230000014509 gene expression Effects 0.000 description 17
- 241000699666 Mus <mouse, genus> Species 0.000 description 15
- 230000000694 effects Effects 0.000 description 15
- 238000003119 immunoblot Methods 0.000 description 14
- 230000011664 signaling Effects 0.000 description 14
- 210000001519 tissue Anatomy 0.000 description 14
- 230000004913 activation Effects 0.000 description 12
- 238000001514 detection method Methods 0.000 description 11
- OHCQJHSOBUTRHG-KGGHGJDLSA-N FORSKOLIN Chemical compound O=C([C@@]12O)C[C@](C)(C=C)O[C@]1(C)[C@@H](OC(=O)C)[C@@H](O)[C@@H]1[C@]2(C)[C@@H](O)CCC1(C)C OHCQJHSOBUTRHG-KGGHGJDLSA-N 0.000 description 10
- 108010017324 STAT3 Transcription Factor Proteins 0.000 description 10
- 102100024040 Signal transducer and activator of transcription 3 Human genes 0.000 description 10
- 239000003112 inhibitor Substances 0.000 description 10
- 102000004877 Insulin Human genes 0.000 description 9
- 108090001061 Insulin Proteins 0.000 description 9
- 210000000577 adipose tissue Anatomy 0.000 description 9
- 230000037396 body weight Effects 0.000 description 9
- 229940125396 insulin Drugs 0.000 description 9
- 230000002829 reductive effect Effects 0.000 description 9
- 230000008685 targeting Effects 0.000 description 9
- 102000017946 PGC-1 Human genes 0.000 description 8
- 108700038399 PGC-1 Proteins 0.000 description 8
- 238000000338 in vitro Methods 0.000 description 8
- 235000021590 normal diet Nutrition 0.000 description 8
- 238000007492 two-way ANOVA Methods 0.000 description 8
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- 230000002503 metabolic effect Effects 0.000 description 7
- 241001465754 Metazoa Species 0.000 description 6
- 102000023984 PPAR alpha Human genes 0.000 description 6
- 210000001185 bone marrow Anatomy 0.000 description 6
- 239000000284 extract Substances 0.000 description 6
- 230000001404 mediated effect Effects 0.000 description 6
- 238000003753 real-time PCR Methods 0.000 description 6
- SUZLHDUTVMZSEV-UHFFFAOYSA-N Deoxycoleonol Natural products C12C(=O)CC(C)(C=C)OC2(C)C(OC(=O)C)C(O)C2C1(C)C(O)CCC2(C)C SUZLHDUTVMZSEV-UHFFFAOYSA-N 0.000 description 5
- 108010028924 PPAR alpha Proteins 0.000 description 5
- OHCQJHSOBUTRHG-UHFFFAOYSA-N colforsin Natural products OC12C(=O)CC(C)(C=C)OC1(C)C(OC(=O)C)C(O)C1C2(C)C(O)CCC1(C)C OHCQJHSOBUTRHG-UHFFFAOYSA-N 0.000 description 5
- 238000005265 energy consumption Methods 0.000 description 5
- 235000013305 food Nutrition 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 210000002966 serum Anatomy 0.000 description 5
- 230000019491 signal transduction Effects 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- 230000000476 thermogenic effect Effects 0.000 description 5
- 238000011870 unpaired t-test Methods 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- YASAKCUCGLMORW-UHFFFAOYSA-N Rosiglitazone Chemical compound C=1C=CC=NC=1N(C)CCOC(C=C1)=CC=C1CC1SC(=O)NC1=O YASAKCUCGLMORW-UHFFFAOYSA-N 0.000 description 4
- 230000003044 adaptive effect Effects 0.000 description 4
- 210000000593 adipose tissue white Anatomy 0.000 description 4
- 230000037406 food intake Effects 0.000 description 4
- 235000012631 food intake Nutrition 0.000 description 4
- 238000007446 glucose tolerance test Methods 0.000 description 4
- 210000002865 immune cell Anatomy 0.000 description 4
- 230000001965 increasing effect Effects 0.000 description 4
- CGIGDMFJXJATDK-UHFFFAOYSA-N indomethacin Chemical compound CC1=C(CC(O)=O)C2=CC(OC)=CC=C2N1C(=O)C1=CC=C(Cl)C=C1 CGIGDMFJXJATDK-UHFFFAOYSA-N 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 238000001543 one-way ANOVA Methods 0.000 description 4
- 230000035924 thermogenesis Effects 0.000 description 4
- 239000013598 vector Substances 0.000 description 4
- 230000003442 weekly effect Effects 0.000 description 4
- APIXJSLKIYYUKG-UHFFFAOYSA-N 3 Isobutyl 1 methylxanthine Chemical compound O=C1N(C)C(=O)N(CC(C)C)C2=C1N=CN2 APIXJSLKIYYUKG-UHFFFAOYSA-N 0.000 description 3
- 102100033295 Glial cell line-derived neurotrophic factor Human genes 0.000 description 3
- 101000974934 Homo sapiens Cyclic AMP-dependent transcription factor ATF-2 Proteins 0.000 description 3
- 101000997829 Homo sapiens Glial cell line-derived neurotrophic factor Proteins 0.000 description 3
- 101150104581 Il27ra gene Proteins 0.000 description 3
- 101150040067 STK11 gene Proteins 0.000 description 3
- 230000033228 biological regulation Effects 0.000 description 3
- 210000002798 bone marrow cell Anatomy 0.000 description 3
- 230000036757 core body temperature Effects 0.000 description 3
- 210000001671 embryonic stem cell Anatomy 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 238000000684 flow cytometry Methods 0.000 description 3
- 238000003205 genotyping method Methods 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 210000004185 liver Anatomy 0.000 description 3
- 238000001325 log-rank test Methods 0.000 description 3
- 230000036284 oxygen consumption Effects 0.000 description 3
- 102000002574 p38 Mitogen-Activated Protein Kinases Human genes 0.000 description 3
- 108010068338 p38 Mitogen-Activated Protein Kinases Proteins 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 238000007619 statistical method Methods 0.000 description 3
- 238000007920 subcutaneous administration Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 230000009885 systemic effect Effects 0.000 description 3
- JWZZKOKVBUJMES-UHFFFAOYSA-N (+-)-Isoprenaline Chemical compound CC(C)NCC(O)C1=CC=C(O)C(O)=C1 JWZZKOKVBUJMES-UHFFFAOYSA-N 0.000 description 2
- 108700028369 Alleles Proteins 0.000 description 2
- 238000011746 C57BL/6J (JAX™ mouse strain) Methods 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- 108700024394 Exon Proteins 0.000 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 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- 101710113864 Heat shock protein 90 Proteins 0.000 description 2
- 102100034051 Heat shock protein HSP 90-alpha Human genes 0.000 description 2
- 101001046686 Homo sapiens Integrin alpha-M Proteins 0.000 description 2
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 102100022338 Integrin alpha-M Human genes 0.000 description 2
- 101150051655 Lyz2 gene Proteins 0.000 description 2
- 101100339600 Mus musculus Hprt1 gene Proteins 0.000 description 2
- 102000000536 PPAR gamma Human genes 0.000 description 2
- 108010016731 PPAR gamma Proteins 0.000 description 2
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 2
- 238000002105 Southern blotting Methods 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 239000012190 activator Substances 0.000 description 2
- 230000011759 adipose tissue development Effects 0.000 description 2
- 238000003149 assay kit Methods 0.000 description 2
- 210000002459 blastocyst Anatomy 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000000749 co-immunoprecipitation Methods 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 230000002950 deficient Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 2
- 229960003957 dexamethasone Drugs 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 238000004520 electroporation Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 201000010063 epididymitis Diseases 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 210000004602 germ cell Anatomy 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 238000011532 immunohistochemical staining Methods 0.000 description 2
- 238000003364 immunohistochemistry Methods 0.000 description 2
- 229960000905 indomethacin Drugs 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 229940039009 isoproterenol Drugs 0.000 description 2
- 238000011813 knockout mouse model Methods 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 210000005228 liver tissue Anatomy 0.000 description 2
- 210000004698 lymphocyte Anatomy 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 210000002540 macrophage Anatomy 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 210000000066 myeloid cell Anatomy 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 230000026731 phosphorylation Effects 0.000 description 2
- 238000006366 phosphorylation reaction Methods 0.000 description 2
- 210000000229 preadipocyte Anatomy 0.000 description 2
- 230000009822 protein phosphorylation Effects 0.000 description 2
- 230000006798 recombination Effects 0.000 description 2
- 238000005215 recombination Methods 0.000 description 2
- 229960004586 rosiglitazone Drugs 0.000 description 2
- -1 small molecule compound Chemical class 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 210000004003 subcutaneous fat Anatomy 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000001960 triggered effect Effects 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- 102000011690 Adiponectin Human genes 0.000 description 1
- 108010076365 Adiponectin Proteins 0.000 description 1
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 1
- 241000237519 Bivalvia Species 0.000 description 1
- 238000011740 C57BL/6 mouse Methods 0.000 description 1
- 101100026251 Caenorhabditis elegans atf-2 gene Proteins 0.000 description 1
- 108010077544 Chromatin Proteins 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 102000029816 Collagenase Human genes 0.000 description 1
- 108060005980 Collagenase Proteins 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 1
- 241001559542 Hippocampus hippocampus Species 0.000 description 1
- 102100024594 Histone-lysine N-methyltransferase PRDM16 Human genes 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 1
- 101000686942 Homo sapiens Histone-lysine N-methyltransferase PRDM16 Proteins 0.000 description 1
- 101001128431 Homo sapiens Myeloid-derived growth factor Proteins 0.000 description 1
- 102000003777 Interleukin-1 beta Human genes 0.000 description 1
- 108090000193 Interleukin-1 beta Proteins 0.000 description 1
- 102000014158 Interleukin-12 Subunit p40 Human genes 0.000 description 1
- 108010011429 Interleukin-12 Subunit p40 Proteins 0.000 description 1
- 108090000978 Interleukin-4 Proteins 0.000 description 1
- 108010002616 Interleukin-5 Proteins 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- 102000016267 Leptin Human genes 0.000 description 1
- 108010092277 Leptin Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- 102000043136 MAP kinase family Human genes 0.000 description 1
- 108091054455 MAP kinase family Proteins 0.000 description 1
- 101100220687 Mus musculus Cidea gene Proteins 0.000 description 1
- 101100018698 Mus musculus Il27 gene Proteins 0.000 description 1
- 101100154912 Mus musculus Tyrp1 gene Proteins 0.000 description 1
- NPGIHFRTRXVWOY-UHFFFAOYSA-N Oil red O Chemical compound Cc1ccc(C)c(c1)N=Nc1cc(C)c(cc1C)N=Nc1c(O)ccc2ccccc12 NPGIHFRTRXVWOY-UHFFFAOYSA-N 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 208000025174 PANDAS Diseases 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 208000021155 Paediatric autoimmune neuropsychiatric disorders associated with streptococcal infection Diseases 0.000 description 1
- 240000000220 Panda oleosa Species 0.000 description 1
- 235000016496 Panda oleosa Nutrition 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 229940122907 Phosphatase inhibitor Drugs 0.000 description 1
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 108010044012 STAT1 Transcription Factor Proteins 0.000 description 1
- 101100403108 Schizosaccharomyces pombe (strain 972 / ATCC 24843) mud1 gene Proteins 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 102100029904 Signal transducer and activator of transcription 1-alpha/beta Human genes 0.000 description 1
- 238000000692 Student's t-test Methods 0.000 description 1
- AUYYCJSJGJYCDS-LBPRGKRZSA-N Thyrolar Chemical compound IC1=CC(C[C@H](N)C(O)=O)=CC(I)=C1OC1=CC=C(O)C(I)=C1 AUYYCJSJGJYCDS-LBPRGKRZSA-N 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 1
- 101150022052 UCP1 gene Proteins 0.000 description 1
- 238000005411 Van der Waals force Methods 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 238000000540 analysis of variance Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 238000009529 body temperature measurement Methods 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 210000003483 chromatin Anatomy 0.000 description 1
- 235000020639 clam Nutrition 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 229960002424 collagenase Drugs 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000010219 correlation analysis Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000016396 cytokine production Effects 0.000 description 1
- 238000013480 data collection Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 235000020931 dietary conditions Nutrition 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000003828 downregulation Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethanethiol Chemical compound CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 description 1
- 238000010195 expression analysis Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000010363 gene targeting Methods 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 238000005469 granulation Methods 0.000 description 1
- 230000003179 granulation Effects 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 230000013632 homeostatic process Effects 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 102000056374 human MYDGF Human genes 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 238000012151 immunohistochemical method Methods 0.000 description 1
- 229940088592 immunologic factor Drugs 0.000 description 1
- 239000000367 immunologic factor Substances 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 229940039781 leptin Drugs 0.000 description 1
- NRYBAZVQPHGZNS-ZSOCWYAHSA-N leptin Chemical compound O=C([C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CC(C)C)CCSC)N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CS)C(O)=O NRYBAZVQPHGZNS-ZSOCWYAHSA-N 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 230000004132 lipogenesis Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 235000020938 metabolic status Nutrition 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 230000017513 negative regulation of heat generation Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 206010033675 panniculitis Diseases 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 1
- 108091008725 peroxisome proliferator-activated receptors alpha Proteins 0.000 description 1
- 230000009038 pharmacological inhibition Effects 0.000 description 1
- 230000037081 physical activity Effects 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000001698 pyrogenic effect Effects 0.000 description 1
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 238000000528 statistical test Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 210000003207 subcutaneous adipocyte Anatomy 0.000 description 1
- 101150074703 sun gene Proteins 0.000 description 1
- 230000009469 supplementation Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000012353 t test Methods 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 1
- 229940035722 triiodothyronine Drugs 0.000 description 1
- 238000012762 unpaired Student’s t-test Methods 0.000 description 1
- 210000000689 upper leg Anatomy 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 210000000636 white adipocyte Anatomy 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
- A01K67/0275—Genetically modified vertebrates, e.g. transgenic
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/177—Receptors; Cell surface antigens; Cell surface determinants
- A61K38/1793—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
- A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/715—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons
- C07K14/7155—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons for interleukins [IL]
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
- G01N33/5041—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects involving analysis of members of signalling pathways
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/07—Animals genetically altered by homologous recombination
- A01K2217/075—Animals genetically altered by homologous recombination inducing loss of function, i.e. knock out
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/20—Animal model comprising regulated expression system
- A01K2217/203—Animal model comprising inducible/conditional expression system, e.g. hormones, tet
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/20—Animal model comprising regulated expression system
- A01K2217/206—Animal model comprising tissue-specific expression system, e.g. tissue specific expression of transgene, of Cre recombinase
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2500/00—Screening for compounds of potential therapeutic value
- G01N2500/02—Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Immunology (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Urology & Nephrology (AREA)
- Gastroenterology & Hepatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Biochemistry (AREA)
- Cell Biology (AREA)
- Diabetes (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Obesity (AREA)
- Food Science & Technology (AREA)
- General Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Endocrinology (AREA)
- Rheumatology (AREA)
Abstract
本发明公开用于控制生物体内热量产生的组合物、方法及用途,本发明的组合物包含用于调节EBI‑3和p28结合或两者的二聚体与其受体的结合、或者下游信号通路的分子。本发明揭示了IL27信号在调节热量产生中的作用。分子机制研究证实其直接作用于脂肪细胞,激活p38MAPK‑PGC1信号并刺激UCP1的产生。IL‑27的治疗性给药逆转EBI‑3KO小鼠低温诱导的低体温症。因此,本发明揭示IL‑27信号在能量代谢过程中的关键作用,为控制体温提供了一种可行方案。
Description
技术领域
本发明涉及生物技术领域,具体地涉及组合物、方法及用途,尤其涉及用于控制生物体内热量产生的组合物、方法及用途。
背景技术
棕色和米色脂肪产热在维持体温的发展方面发挥着关键作用。尽管棕色/米色脂肪的定向和激活已为人们所知,但是脂肪组织中免疫因子的多样性和丰富性尚不清楚。
US20170128410A1公开了一种用于实现哺乳动物能量平衡的方法和组合物,尤其是在脂肪生成抑制的过程中使用福司柯林实现哺乳动物能量平衡的方法,其中将福司柯林分开添加到分化前的前脂肪细胞以及还添加到成熟脂肪细胞以比较地评估脂肪生成抑制潜力,所述过程包括在由未分化的前脂肪细胞组成的预接种微板中添加一定量浓度的福司柯林的步骤。
熊燕在Lkb1调控棕色脂肪组织的功能及机制研究中,采用Adipoq-Cre/Loxp***建立脂肪组织特异Lkb1KO小鼠模型(Ad-Lkb1),从组织形态学、分子水平及体内外实验相结合研究Lkb1调控棕色脂肪产热及机体能量代谢的影响,并且通过染色质免疫共沉淀、蛋白免疫共沉淀及双荧光素酶报告分析,揭示Lkb1调控机体能量代谢及Ucp1表达的分子机制。
背景技术中的信息仅仅在于说明本发明的总体背景,不应视为承认或以任何形式暗示这些信息构成本领域一般技术人员所公知的现有技术。
发明内容
为解决现有技术中的至少部分技术问题,本发明提供用于控制生物体内热量产生的组合物、方法及用途。本发明研究发现(EBI-3/p28)信号在改善体内热量产生方面具有重要作用,证实IL-27直接作用于脂肪细胞,激活p38MAPK-PGC1信号并刺激UCP1的产生。具体地,本发明包括以下内容。
本发明的第一方面,提供一种用于控制生物体内热量产生的组合物,其包含用于调节EBI-3和p28结合或两者的二聚体与其受体的结合、或者下游信号通路的分子。
根据本发明所述的组合物,优选地,所述调节是指抑制或降低EBI-3和p28的结合,或者抑制或降低两者的二聚体与其受体的结合,或者下调下游信号通路,且所述控制是指抑制热量产生,进而降低所述生物体的温度。
根据本发明所述的组合物,优选地,所述分子包括生物大分子或小分子化合物,如抗EBI-3抗体、抗p28抗体、抗IL-27R抗体、EBI-3可溶性片段、p28可溶性片段、IL-27R可溶性片段等。
根据本发明所述的组合物,优选地,所述调节是指促进或增强EBI-3和p28的结合,或者促进或增强两者的二聚体与其受体的结合,或者上调下游信号通路,且所述控制是指促进热量产生,进而提高所述生物体的温度。
根据本发明所述的组合物,优选地,所述分子包括重组IL-27蛋白及其修饰物、具备激活IL-27R活性的多肽或小分子化合物。
本发明的第二方面,提供一种用于控制生物体内热量产生的方法,向所述生物体施用用于调节EBI-3和p28结合或两者的二聚体与其受体的结合、或者下游信号通路的分子。
根据本发明所述的用于控制生物体内热量产生的方法,优选地,所述生物体包括人类受试者或其组织或细胞、哺乳动物或其组织或细胞、或者微生物。
根据本发明所述的用于控制生物体内热量产生的方法,优选地,所述组织或细胞为体外组织或细胞。
本发明的第三方面,提供一种用于筛选能够控制生物体内热量产生的候选化合物的方法,其包括测量施用所述待测化合物前后,EBI-3和p28结合情况,或两者的二聚体与其受体的结合情况、或者由于所述结合引发的下游信号通路的步骤。
根据本发明所述的用于筛选能够控制生物体内热量产生的候选化合物的方法,优选地,当施用所述待测化合物后,EBI-3和p28的结合被抑制或降低、或者两者的二聚体与其受体的结合被抑制或降低,或者由于所述结合引发的下游信号被下调时,将所述待测化合物鉴定为抑制生物体内热量产生的候选化合物。
根据本发明所述的用于筛选能够控制生物体内热量产生的候选化合物的方法,优选地,当施用所述待测化合物后,EBI-3和p28的结合被促进或增强、或者两者的二聚体与其受体的结合被促进或增强,或者由于所述结合引发的下游信号被上调时,将所述待测化合物鉴定为促进生物体内热量产生的候选化合物。
根据本发明所述的用于筛选能够控制生物体内热量产生的候选化合物的方法,优选地,包括以下步骤:
(1)测量生物体模型内EBI-3和p28结合的参数,或两者的二聚体与其受体结合的参数、或者下游信号通路活化情况的参数,得到第一测量值的步骤;
(2)向所述生物体模型施用待测化合物后,测量生物体模型内EBI-3和p28结合的参数,或两者的二聚体与其受体结合的参数、或者下游信号通路活化情况的参数,得到第二测量值的步骤;和
(3)比较第一测量值和第二测量值的步骤。
根据本发明所述的用于筛选能够控制生物体内热量产生的候选化合物的方法,优选地,包括以下步骤:
(1)测量第一生物体模型内EBI-3和p28结合的参数,或两者的二聚体与其受体结合的参数、或者下游信号通路活化情况的参数,得到第一测量值的步骤;
(2)测量第二生物体模型内EBI-3和p28结合的参数,或两者的二聚体与其受体结合的参数、或者下游信号通路活化情况的参数,得到第二测量值的步骤,其中所述第一生物体模型与所述第二生物体模型属于相同的模型,且所述第一生物体模型未施用待测化合物,所述第二生物体模型施用待测化合物;和
(3)比较第一测量值和第二测量值的步骤。
根据本发明所述的用于筛选能够控制生物体内热量产生的候选化合物的方法,优选地,包括以下步骤:
(1)测量施用待测化合物后生物体模型内EBI-3和p28结合的参数,或两者的二聚体与其受体结合的参数、或者下游信号通路活化情况的参数,得到测量值的步骤;
(2)将所述测量值与参考值比较的步骤,其中,所述参考值为由生物体模型在未施用待测化合物时测量得到的EBI-3和p28结合的参数,或两者的二聚体与其受体结合的参数、或者下游信号通路活化情况的参数。
根据本发明所述的用于筛选能够控制生物体内热量产生的候选化合物的方法,优选地,所述生物体模型包括动物模型或细胞模型。
本发明的第四方面,提供第一方面的组合物在制备用于代谢疾病制剂中的用途。
本发明揭示了IL-27(EBI-3/p28)信号在改善体内热量产生中的作用。证实IL-27直接作用于脂肪细胞,激活p38 MAPK-PGC1信号并刺激UCP1的产生。IL-27的治疗性给药可逆转EBI-3KO小鼠低温诱导的低体温症。
附图说明
图1为IL-27信号促进热量产生和能量消耗的图。
图2为IL-27直接靶向脂肪细胞以促进热量产生的图。
图3为IL-27促进热量产生的图。
图4显示IL-27信号缺陷小鼠的能量消耗和热量产生减少。
图5显示IL-27Rα信号的产热作用不是通过直接作用于CD2+淋巴细胞或Lyz2+骨髓细胞。
图6为在HFD诱导的适应性热产生作用的IL-27RαKO和WT嵌合体的表型。
图7显示IL-27上调UCP1并改善皮下白色脂肪组织的褐变。
图8显示IL-27促进热量生产作用。
具体实施方式
现详细说明本发明的多种示例性实施方式,该详细说明不应认为是对本发明的限制,而应理解为是对本发明的某些方面、特性和实施方案的更详细的描述。
应理解本发明中所述的术语仅仅是为描述特别的实施方式,并非用于限制本发明。另外,对于本发明中的数值范围,应理解为具体公开了该范围的上限和下限以及它们之间的每个中间值。在任何陈述值或陈述范围内的中间值以及任何其他陈述值或在所述范围内的中间值之间的每个较小的范围也包括在本发明内。这些较小范围的上限和下限可独立地包括或排除在范围内。
除非另有说明,否则本文使用的所有技术和科学术语具有本发明所述领域的常规技术人员通常理解的相同含义。虽然本发明仅描述了优选的方法和材料,但是在本发明的实施或测试中也可以使用与本文所述相似或等同的任何方法和材料。本说明书中提到的所有文献通过引用并入,用以公开和描述与所述文献相关的方法和/或材料。在与任何并入的文献冲突时,以本说明书的内容为准。除非另有说明,否则“%”为基于重量的百分数。
本文所用术语“热量产生”可以术语“热产生”、“热产生作用”和“生热作用”互换使用。在本文中生热作用是指哺乳动物,包括人调节热量产生的过程。
本发明中,EBI-3和p28结合是指EBI-3亚单位和p28亚单位通过相互作用形成具有一定空间排布的异二聚体形式的结构,相互作用可以是以二硫键方式进行连接。
本文中所用术语“受体”是指是指能够同上述二聚体结合并能引起细胞功能变化的生物大分子。其至少含有识别并结合所述二聚体的活性部位和负责产生应答反应的功能活性部位,并由此启动一系列的生化反应,最终导致靶细胞产生生物效应。受体与本发明的二聚体之间结合的结果是受体被激活,并产生受体激活后续信号传递的基本步骤。在生理条件下,受体与所述二聚体之间的结合不通过共价键介导,主要靠离子键、氢键、范德华力和疏水作用而相互结合。
本文所用术语“信号通路”信号通路是指当细胞里要发生某种反应时,信号从细胞外到细胞内传递了一种信息,细胞要根据这种信息来做出反应的现象。本发明的信号通路尤其是指能将细胞外的分子信号经细胞膜传入细胞内发挥效应的一系列酶促反应通路。
[用于控制生物体内热量产生的组合物]
本发明提供一种用于控制生物体内热量产生的组合物,其包含用于调节EBI-3和p28结合或两者的二聚体与其受体的结合、或者下游信号通路的分子。优选地,本发明的组合物包含用于调节IL27信号或IL27与其受体结合或下游信号通路的分子。其中,下游信号通路包括但不限于JAK/STAT3、p38MAPK等。本发明中,产热以及能力消耗由脂肪细胞介导,所述脂肪细胞包括但不限于棕色/米色脂肪细胞以及白色脂肪细胞。在本发明中,上述脂肪细胞作为IL27靶细胞,而不是免疫细胞。
[用于筛选能够控制生物体内热量产生的候选化合物的方法]
本发明中,筛选方法至少包括使用试剂测量施用所述待测化合物前后,EBI-3和p28结合情况,或两者的二聚体与其受体的结合情况、或者由于所述结合引发的下游信号通路的步骤。上述结合情况还包括通过使用试剂测量相关参数的步骤,所述参数包括二者形成的二聚体的量,或是代谢表型,或是产热蛋白的量。其所述产热蛋白包括但不限于UCP1、PPARα,PGC-1α。在本发明中,下游信号通路活化(有时也称为激活)是指下游靶基因的进一步表达。优选地,所述试剂包括SEQIDNO.:1-30所示的序列。
实施例
1.实验材料
1.1材料
1.1.1小鼠
白介素-27RαKO(B6N.129P2-Il27ratm1Mak/J,编号:018078)和EBI-3KO(B6.129X1-Ebi3tm1Rsb/J,编号:008691)小鼠购自Jackson Laboratory。Adipoq-cre小鼠由Yong Liu教授提供。IL-27Rαf/f小鼠[本文]由本实验室制备。动物实验按照暨南大学机构动物保护与利用委员会和耶鲁大学机构动物护理与利用委员会批准的伦理规范和协议进行。所有的实验都使用了性别和年龄匹配的小鼠组,每个笼子里有4到6只动物。随机分组进行IL-27治疗实验和骨髓嵌合体的产生。小鼠组饲养在温度和湿度控制的特定无菌动物设施中,温度为25℃,光照:黑暗周期为12:12h,可自由获得食物和水。
1.1.2试剂
使用Bio-Plex-ProTMHuman Cytokine 17-Plex分析试剂盒(Cat.No.M5000031YV)和Bio-Plex ProTMHuman Inflammation Panel 1(Cat.No.171-AL001M)在Bio-Plex***(BioRad)上分析人血清样品。人IL-27ELISA试剂盒(434607)购自Biolegend。葡萄糖(63005518)购自中国国药化学试剂公司。血糖试纸(one-touch)购自Johnson和Jonhson。抗小鼠pAKT单抗(4060)、抗小鼠AKT单抗(4685)、抗小鼠GAPDH单抗(5174)、抗小鼠HSP90单抗(4877)、抗小鼠PPARγ单抗(2435)、抗小鼠pSTAT3单抗(9145)、抗小鼠STAT3单抗(9139)、抗小鼠p-p38 MAPK(9211)、抗小鼠p38 MAPK(9212)、抗小鼠pATF2单抗(9221)、抗小鼠ATF2单抗(35031)购自Cell Signaling Technology。抗小鼠UCP1单抗(ab10983)、抗小鼠PGC-1α单抗(ab54481)、抗小鼠PPARα单抗(ab8934)、抗小鼠PRDM16单抗(ab202344)、抗小鼠IL-27Rα单抗(ab5997)购自Abcam。APC-CY7-抗小鼠CD45(克隆30-F11)购自BD。APC抗小鼠CD11b(克隆M1/70)、PE抗小鼠F4/80(克隆BM8)购自Sungene Biotech(中国天津)。重组小鼠IL-27(rmIL-27)(577408)购自Biolegend。STAT3抑制剂SH-4-54和p38 MAPK抑制剂SB203580购自Selleck Chemicals。棕色/米色脂肪分化剂***(D4902)、罗格列酮(R2408)、吲哚美辛(I7378)、异丙肾上腺素(I5627)、福司可林(F6886)、3-异丁基-1-甲基黄嘌呤(IBMX)(I5879)、甘油三酯检测试剂盒(T2449)、胰岛素(I3536)购自Sigma。瘦素检测试剂盒(MOB00)、脂联素检测试剂盒(MRP300)购自R和D。胰岛素检测试剂盒(90080)购自CrystalChem。
2.实验方法
2.1酶联免疫吸附试验
如上所述获得人血清。在喂食HFD 10周后收集小鼠血清。人IL-27和小鼠胰岛素使用ELISA试剂盒按使用说明书进行检测。
2.2 Luminex免疫分析法
如上所述采集人血清。使用Bio-Plex-ProTMHuman Cytokine 17-Plex分析试剂盒(Cat.No.M5000031YV)和Bio-Plex ProTMHuman Inflammation Panel1(Cat.No.171-AL001M)在Bio-Plex 200流式液相芯片分析***(BioRad)上按照用户手册的说明进行人血清炎症因子的检测。
2.3蛋白质印迹
使用补充有完整的蛋白酶抑制剂(Roche)和磷酸酶抑制剂Cocktail2(P5726,Sigma)的RIPA裂解缓冲液(Beyotime,China)提取全细胞裂解液或组织裂解液,并使用上清液进行后续分析。蛋白质在Loading染料(BL502A,Bio-sharp)中稀释,在95℃加热℃ 10分钟,在4-12%聚丙烯酰胺凝胶上跑胶。将蛋白质转移到聚偏氟乙烯膜上,并用图中提到的商业抗体和方法-试剂进行蛋白质印迹实验。
2.4组织学
将脂肪或肝组织在4℃下在4%多聚甲醛/1×PBS中固定过夜并在切片之前包埋在石蜡中。切片用苏木精和伊红(H和E)或油红O(liver)染色并在亮视野显微镜下照相。
2.5免疫组织化学
根据制造商的说明,使用兔特异性HRP/DAB(ABC)检测IHC试剂盒(ab64261,Abcam)进行UCP1和免疫组化。
2.6核心体温测量和冷刺激实验
在温度控制的冷室内进行冷刺激实验。将小鼠在4℃单独放置在单独的笼子(没有垫底)中。小鼠可自由获得食物和水。每2小时使用数字温度计和直肠热电偶探头(TH-212,HICHANCE,China)记录直肠核心体温。如果核心体温降到20℃以下,则将个体小鼠安乐死,并作为存活分析的事件进行评分。
2.7代谢笼
使用耶鲁大学环境控制啮齿动物培养箱内的综合实验室动物监测***(CLAMS,Columbus Instruments,Columbus,OH)代谢笼测定小鼠的能量损耗和能量消耗。在数据收集之前,将小鼠单独饲养并在代谢室中适应48小时。老鼠可以自由获得食物和水。对每只老鼠进行体力活动的连续监测和食物摄入。在实验期间,每只小鼠每小时收集4次CO2/O2浓度。
2.8RNA提取及基因表达分析
使用TRNzol通用试剂(Tiangen,北京)从冷冻组织中提取总RNA,使用Nanodrop2000紫外可见分光光度计(Thermo)进行定量。cDNA采用PrimeS criptTMRT Reagent试剂盒(TAKARA,大连)用1μg总RNA经逆转录聚合酶链反应检测制备得到。cDNA实时定量PCR采用TBGreenTM Premix-EX-TaqTM II试剂盒(TAKARA)在CFX96TM实时PCR检测***(Bio-Rad)上进行。以小鼠HPRT为内参照,通过△△Ct法采用免疫组化方法计算表达中倍数变化。引物对序列如下。小鼠HPRT:forward-TCATTATGCCGAGGATTTG(SEQIDNO.1),reverse-GCCTCCCATCTCCTTCAT(SEQIDNO.2);TNF-α:forw ard-CTACTGAACTTCGGGGTGAT(SEQIDNO.3),reverse-CAGGCTTGTCACTCGAATT(SEQIDNO.4);IL-6:forward-TCCAGTTGCCTTCTTGGGAC(SEQIDNO.5),reverse-GTGTAATTAAGCGCCGACTTG(SEQIDNO.6);IL-12p40:forward-GGCTGGTGCAAAGAAACATGGACTTGA(SEQIDNO.7),reverse-TGCAGACAGAGACGCCATTCCACAT(SEQIDNO.8);IL-1β:forward-CAACCAACAAGTGATATTCTCCATG(SEQIDNO.9),reverse-GATCCACACTCTCCAGCTGCA(SEQIDNO.10);IL-4:forward-GAAAACTCCATGCTTGAAGAA(SEQIDNO.11),reverse-TCTTTCAGTGATGTGGACTTG(SEQIDNO.12),IL-5:forward-TCACCGAGCTCTGTTGACAA(SEQIDNO.13),reverse-CCACACTTCTCTTTTTGGCG(SEQIDNO.14);IL-13:forward-TGAGCAACATCACACAAGACC(SEQIDNO.15),reverse-GGCCTTGCGGTTACAGAGG(SEQIDNO.16);Ucp1:forward-ACTGCCACACCTCCAGTCATT(SEQIDNO.17),reverse-CTTTGCCTCACTCAGGATTGG(SEQIDNO.18);Cox8b:forward-GAACCATGAAGCCAACGACT(SEQIDNO.19),reverse-GCGAAGTTCACAGTGGTTCC(SEQIDNO.20);Cidea:forward-TGCTCTTCTGTATCGCCCAGT(SEQIDNO.21),reverse-GCCGTGTTAAGGAATCTGCTG(SEQIDNO.22);Prdm16:forward-CAGCACGGTGAAGCCATTC(SEQIDNO.23),reverse-GCGTGCATCCGCTTGTG(SEQIDNO.24);Adiponectin:forward-GAATCATTATGACGGCAGCA(SEQIDNO.25),reverse-TCATGTACACCGTGATGTGGTA(SEQIDNO.26);Elovl3:forward-TCCGCGTTCTCATGTAGGTCT(SEQIDNO.27),reverse-GGACCTGATGCAACCCTATGA(SEQIDNO.28)。
2.9骨髓嵌合体
以10周龄的IL-27RαKO和WT小鼠作为骨髓细胞供体。从后肢股骨分离骨髓,经红细胞溶解后,通过细胞过滤器(40μm)过滤,用无菌PBS洗涤细胞球3次,计数并保存于冰上以备注射。对10周龄的IL-27RαKO和WT小鼠进行致死性辐射(900rads),经眼静脉注射移植1x107骨髓细胞,将小鼠置于特定的无病原体设施中,补充消毒水和饲料8周,以重建免疫***,然后进行代谢研究。
2.10原代米色脂肪细胞制备
将腹股沟皮下脂肪组织切碎,于含有胶原酶II(1mg/ml)的PBS中37℃消化45分钟,组织悬浮液通过100μm细胞滤器过滤,以600g离心5min,使基质血管部分(SVF)颗粒化。颗粒化后进一步通过40μm细胞滤器过滤,并将其置于胶原涂层板上。过夜培养后,去除含有未粘附细胞的上清液。前体脂肪细胞在含10%FBS+胰岛素(5μg/ml)的DMEM中生长融合。使用***(1μM),3-异丁基-1-甲基黄嘌呤(IBMX,0.5mM),胰岛素(5μg/ml),吲哚美辛(125nM)和罗格列酮(1μM)处理2天诱导融合细胞分化,然后使用胰岛素(5μg/ml)和三碘甲状腺原氨酸(T3,1nM)单独处理另外的5天。在第7天,细胞在有和没有IL-27(100ng/ml)的情况下预处理过夜,然后用异丙肾上腺素(10μM)或forskolin(10μM)处理4-6小时。第7天加入IL-27(100ng/ml)处理0-120min用于蛋白磷酸化分析,或12-24h用于蛋白表达水平检测。对于信号抑制实验,STAT3抑制剂(C188-9,10μM)或p38 MAPK抑制剂(SB203580,10μM)在IL-27治疗前0.5h和治疗期间加入培养基。
2.11流式细胞分析(FACS)。
如上所述,将附睾脂肪组织切碎并消化。SVF颗粒用于APC-CY7抗小鼠CD45、APC抗小鼠CD11b、PE抗小鼠F4/80的表面染色,孵育15min后,用PBS洗涤细胞,用BD-FACSVerse流式细胞仪(BD)分析。
2.12产生IL-27Rα条件敲除小鼠。利用基因打靶技术进行IL-27Rα条件敲除小鼠的开发工作。简而言之,利用高保真Taq技术从BAC克隆中扩增出一个包含IL27ra基因外显子3和外显子4的997bps CKO区(GenBank NM_016671.3)、一个5.2kb的5'同源臂和一个3kb的3'同源臂,并与重组位点和选择标记一起组装成靶向载体。经多重限制性内切酶切和全序列分析确定最终的靶向载体,并通过电穿孔法将其转化为129s小鼠胚胎干细胞。鉴定了正确的胚胎干细胞克隆,并将其注射到C57BL/6J小鼠囊胚中。明确鉴定出嵌合体小鼠后,与C57BL/6J小鼠回交。基因分型PCR证实了种系传播。然后将具有IL-27Rαf/w基因型的小鼠与B6 adipoq-cre小鼠饲养。实验选用期望基因型小鼠。检测flox或wt带的基因分型引物:forward:CTGGTTCTGGTATGGTTTGGGGTT(SEQIDNO.29),reverse:TGAAAGAACTCAACAGTGGGCCGG(SEQIDNO.30)。
2.13 IL-27治疗
8周龄的WT小鼠HFD喂养32周并随机分为2组。小鼠腹腔注射rmI L-27(100μg/kg)或PBS,连续15天,然后进行代谢分析。12月龄EBI-3KO小鼠随机分为2组,腹腔注射rmIL-27(100μg/kg)或PBS,连续7天。然后把老鼠放在4℃冷激发实验。
2.14统计分析
使用Graphpad Prism软件(版本7)进行绘图和统计分析。所有的统计检验都在图表图例中有充分的描述,并且符合方差相似的正态分布的标准。未使用统计方法预先确定样本量。两组间比较采用t检验。对于相关样本的评估数据,进行重复测量方差分析(AVOVA)。对于两组以上的评估,采用单因素方差分析和多重比较。对于两个自变量之间的评估,采用多重比较的双因素方差分析。生存分析采用对数秩检验。相关分析采用线性回归分析。否则数据以平均值±s.e.m.表示,除非另有说明。p<0.05被认为具有统计学意义,表示为*p<0.05,**p<0.01,**p<0.001,NS,不显著。
3.实验结果
图1为IL-27信号促进热量产生和能量消耗的图。a-c.IL-27RαKO小鼠和WT对照小鼠HFD喂食4周后置于代谢笼中饲养。在24小时内(n=7-8)监测食物摄入(图中a所示)、氧气消耗(图中b所示)和能量消耗(图中c所示)。d和e:IL-27RαKO小鼠和WT对照小鼠分别HFD喂食6周,然后进行冷刺激(4℃)。存活曲线如图中(d,n=6-8)所示,直肠温度如图中(e,n=11-15)所示。F和g:采用HFD喂食IL-27RαKO和WT小鼠6周,然后进行25℃(图中f所示)或4℃冷刺激2小时(图中g所示)。收集皮下脂肪(SCW)和棕色脂肪组织(BAT)进行UCP1免疫印迹分析。h-j:以正常饮食喂养的IL-27RαKO和WT小鼠置于25℃温度下或者在4℃冷刺激48小时。收集SCW和BAT进行UCP1免疫印迹分析(图中h所示),i为UCP1免疫组织化学染色结果(比例尺=100μm),j为BAT和SCW组织的H和E染色(比例尺=50μm)。(f-h):用ImageJ定量条带密度并归一化。每个泳道代表一个生物学独立的样品,实验至少重复两次,结果相似。数据是生物独立样本的平均值±s.e.m.,双因素方差分析(a-c和e);对数秩检验(d)。*p<0.05,**p<0.01,***p<0.001。
图2为IL-27直接靶向脂肪细胞以促进热量产生的图。a.用IL-27RαKO或WT小鼠作为供体或受体,喂养HFD 10周产生四组骨髓嵌合体。每周记录体重(n=10-13)。b.冷刺激对正常饲料喂养的嵌合小鼠直肠温度的影响(4℃,n=6-7)。c.喂食正常食物的嵌合小鼠在4℃冷刺激12小时后收集SCW进行免疫印迹。d.8周龄的Adipoq-CreIl27raf/f小鼠和Il27raf/f对照组喂食HFD 10周。每周记录体重(n=7-9)。e.冷刺激对正常饮食小鼠直肠温度的影响(4℃,n=16-17)。f.喂食正常食物的Adipoq-Cre Il27raf/f和Il27raf/f小鼠在4℃冷刺激12小时后,收集SCW和BAT进行免疫印迹。g.8周龄的Ucp1-CreIl27raf/f小鼠和Il27raf/f对照组喂食HFD 10周。每周记录体重(n=5-6)。h.冷刺激对正常饮食小鼠直肠温度的影响(4℃,n=5-8)。i.喂食正常食物的Ucp1-CreIl27raf/f和Il27raf/f小鼠在4℃冷刺激12小时后,收集SCW和BAT进行免疫印迹(c,f和i),用ImageJ定量条带密度并归一化。每个泳道代表一个独立的生物样本,实验重复至少两次,结果相似。数据是生物独立样本的平均值±s.e.m.。双因素方差分析(a,b,d,e,g和h)。*p<0.05,**p<0.01,**p<0.001。
图3为IL-27促进热量生产的作用。a.从WTSCW的SVF体外产生原代米色脂肪细胞,然后用rmIL-27(100ng/ml)或PBS处理24小时。裂解细胞,并将蛋白提取物用于免疫印迹分析。b.用rmIL-27(100ng/ml)处理指定时间的WT原代米色脂肪细胞提取物中蛋白质磷酸化的免疫印迹分析。c.来源于WTSCW的原代米色脂肪细胞是在体外产生的。STAT3抑制剂(C188-9,10μM)或p38 MAPK抑制剂(SB203580,10μM)在培养前0.5小时和rmIL-27处理(100ng/ml,持续12小时)持续时间内添加到培养基中。细胞裂解以及蛋白质提取物的代表性免疫印迹如图所示。d-j:用HFD喂食WT小鼠32周,然后腹膜内注射。每隔一天注射rmIL-27(100μg/kg)或PBS注射15天。d.在指定的时间点记录体重(n=8)。e.rmIL-27处理15天后,收集脂肪组织并称重(n=6-8)进行葡萄糖耐量试验(f,n=8)和胰岛素耐量测试(g,n=8)。h.将小鼠禁食过夜,然后腹膜内注射0.75U/kg的胰岛素(15分钟)。进行附睾脂肪中pAKT(pSer473)的免疫印迹分析。i.rmIL-27治疗15天后进行油红O肝组织染色。比例尺=50μm。j-m:UCP1-KO小鼠用HFD喂养20周,然后每隔一天腹腔注射rmIL-27(100μg/kg)或PBS,连续15天。j.在指定的时间点记录体重(n=6)。k.在rmIL-27治疗15天后(n=6)收集脂肪组织和肝脏并称重,在rmIL-27治疗15天后进行GTT(l,n=6)和ITT(m,n=4)试验。用ImageJ定量条带密度并进行归一化(a和c),重复实验至少两次,结果相似。数据是生物独立样本的平均值±s.e.m.。非配对学生t检验(e和k);双因素方差分析(d,f,g,j,l和m)。*p<0.05。
图4显示了IL-27信号缺陷小鼠的能量消耗和热量产生减少。a-c:将8周龄的IL-27RαKO和WT小鼠置于代谢笼中。其中,食物摄取量(a)、氧气消耗量(b)和能量消耗量(c)(n=7-8)如图所示。d.从6-8周龄的WT-ND和IL-27RαKO中分离BAT和SCW组织,切成小块(BAT约0.003g,SCW约0.004g),用Seahorse XF分析仪检测基础耗氧率(n=5只/组,4-5块/组/小鼠)。e.IL-27RαKO和WT小鼠喂食ND对冷刺激的反应存活曲线(4℃,n=9-11)。f.冷刺激对ND喂养小鼠直肠温度的影响(4℃,n=9-11)。g.来自于正常饮食的IL-27RαKO和WT小鼠基因在SCW中的表达采用实时PCR检测(n=9-11)。h-j:EBI-3KO小鼠和以正常食物喂养的WT对照组在4℃冷刺激记录,其存活曲线(h)和直肠温度(i)(n=5-6)如图所示。冷刺激12小时后,收集BAT和SCW,裂解并用于UCP1(j)的免疫印迹分析。用ImageJ定量条带密度,归一化UCP1/HSP90比值。实验进行两次显示结果相似。数据是生物独立样本的平均值±s.e.m.。非配对t检验(d和g);双因素方差分析(a-c,f和i);对数秩检验(e和h)。*p<0.05,**p<0.01,**p<0.001。
图5显示IL-27Rα信号的产热作用不是通过直接作用于CD2+淋巴细胞或Lyz2+骨髓细胞。其中,a.Il27raflox/flox小鼠产生的示意模型。Il27ra位点(top)被靶向载体(second)靶向,该靶向载体包含Il27ra的同源序列,包括外显子3和4两侧的两个LoxP位点和一个Neo选择盒。随后通过电穿孔将线性化载体递送至胚胎干细胞(C57BL/6),随后进行药物选择、PCR筛选和Southern印迹确认。同源重组产生了floxed等位基因(Third)。在通过Southern印迹确定正确靶向ES克隆后,将所选克隆用于囊胚显微注射以产生F0代。经鉴定,将F0与Flp-deleter杂交,确认F1为种系传代以删除Neo盒(Forth)。Cre重组后,floxedIl27ra等位基因的错配将导致外显子3和4的缺失(bottom)。b.Il27raf/f小鼠的基因分型。c.采用实时荧光定量PCR方法检测Il27raf/f和WT小鼠脾脏中Il27ra基因的表达(n=3)。d.用实时荧光定量PCR(n=3,左)检测Il27raf/f和Cd2-Cre Il27raf/f小鼠脾CD3 T细胞中Il27ra基因的表达。将乙硫醇酸盐腹腔注射到Il27raf/f和Lyz2-Cre Il27raf/f小鼠体内4天,然后收集腹腔巨噬细胞,通过实时PCR检测Il27ra基因的表达(n=5-6,右图)。e-h:8周龄的Il27raf/f、Cd2-Cre-Il27raf/f和Lyz2-Cre-Il27raf/f小鼠喂养HFD 10周。其中,e.每周记录体重(n=11-21)。在HFD处理10周后进行GTT(f)和ITT(g)试验(n=11-19)。h.在HFD处理10周后收集脂肪组织并称重(n=11-21)。数据是生物独立样本的平均值±s.e.m.。非配对t检验(c,d);双因素方差分析(e-g);单因素方差分析(h)*p<0.05,***p<0.001。
图6为在HFD诱导的适应性热产生作用的IL-27RαKO和WT嵌合体的表型。将来自CD45.1 WT株的骨髓细胞转到辐照后的CD45.2 IL-27RαKO或CD45.2 WT宿主(1x107细胞/小鼠)产生嵌合体。小鼠安置8周以重建免疫***。如图所示,从不同组织中分离免疫细胞,通过流式细胞仪分析在CD4+T(顶部)中供体(CD45.1+)和宿主(CD45.2+)细胞、CD19+B(中部)或F4/80+巨噬细胞(底部)中的百分比(n=3)。b-d:如图中a所示得到骨髓嵌合体并喂食HFD。经过10周的HFD处理后进行GTT(b,n=10-13)和ITT(c,n=5-7)试验。d.在HFD处理10周后收集脂肪组织和肝脏并称重(n=4-6)。e-g.喂食正常食物的嵌合小鼠在4℃冷刺激12小时,收集SCW和BAT进行组织学分析(e,H和E,比例尺=100μm)或UCP1免疫组化染色(f,比例尺=100μm)。进行SCW的基因表达的实时PCR分析(g,n=12)。数据是生物独立样本的平均值±s.e.m.。非配对t检验(a,f和g);单因素方差分析(d);双因素方差分析(b和c)。*p<0.05,**p<0.01,**p<0.001。
图7显示IL-27上调UCP1并改善皮下白色脂肪组织的褐变。a-c:用HFD喂养8周龄的Il27raf/f和Ucp1-CreIl27raf/f小鼠10周。在HFD处理10周后进行GTT(a)和ITT(b)(n=5-6)试验,收集脂肪组织和肝脏并称重(c,n=5-6)。d.喂食正常食物的Il27raf/f和Ucp1-CreIl27raf/f小鼠在4℃冷刺激12小时后,收集SCW和BAT进行组织学分析(H和E)。比例尺=100μm。e.从WTSCW的SVF中体外产生原代米色脂肪细胞,然后用rmIL-27(100ng/ml)或PBS处理24小时。细胞裂解,蛋白提取物用于免疫印迹分析。WT小鼠的BAT组织作为阳性对照。f.用rmIL-27(100ng/ml)处理指定时间,从WT原代米色脂肪细胞提取物中STAT1磷酸化的免疫印迹分析。g.体外培养WT小鼠皮下脂肪细胞。两种不同的p38 MAPK抑制剂(SB203580,10μM和SB202190,5μM)或三种STAT3抑制剂(C188-9,10μM;Stattic,10μM或HO3867,20μM)在rmIL-27处理前0.5h和处理期间(100ng/ml,12h)加入培养基。免疫印迹法检测UCP1的表达。h-i:来自WT(h)或IL-27RαKO(i)小鼠的SCW的原代米色脂肪细胞经体外培养获得。p38 MAPK抑制剂(SB203580,10μM)或STAT3抑制剂(C188-9,10μM)在rmIL-27处理前0.5h和处理期间(100ng/ml,12h)加入培养基。UCP1或PPARα的表达通过免疫印迹分析。j-m:EBI-3KO小鼠腹腔注射rmIL-27(100μg/kg)或PBS,连续7天,然后在4℃进行冷刺激。j.小鼠对冷刺激的直肠温度(n=6)。冷刺激24小时后SCW和BAT蛋白提取物用于免疫印迹分析。UCP1染色(l)和组织学分析(m)(比例尺=100μm)如图所示。代表性部分如图所示。用ImageJ定量条带密度并归一化。实验重复两次显示结果相似。数据是生物独立样本的平均值±s.e.m.。非配对t检验(c);双因素差分析(a、b和j)*p<0.05,**p<0.01,**p<0.001。
图8显示IL-27促进热量生产的作用。a-i:Il27raf/f和Adipoq-CreIl27raf/f小鼠喂食HFD 32周后每隔一天腹腔注射rmIL-27(100μg/kg)或PBS,共15天。其中,a.采用Bio-RadBio-Plex200多功能分析仪检测血清中的炎症因子。b.指示组织的代表性组织切片(H和E)。c.浸润的CD4或CD8T细胞百分比和肝脏CD4 T细胞的细胞因子的产生用流式细胞仪分析。检测并记录Il27raf/f小鼠的体重(d)、GTT(e)和ITT(f)(n=5-8)。检测并记录Adipoq-CreIl27raf/f小鼠的体重(g)、GTT(h)和ITT(i)(n=4-5)。j-m:用HFD喂养Ucp1-CreIl27raf/f小鼠16周,然后每隔一天腹腔注射rmIL-27(100μg/kg)或PBS,共15天。测定并记录体重(j)、组织重量(k)、葡萄糖耐量试验(l)和胰岛素耐量试验(m)(n=5-6)。数据是生物独立样本的平均值±s.e.m.。单因素方差分析(a和c),双因素方差分析(d-j,l和m);非配对t检验(k)。*p<0.05,**p<0.01,**p<0.001。
4.结论
白色脂肪组织的褐变使小鼠易于增加能量消耗。为了检查全身代谢状况,本发明检测了在正常饮食或HFD处理下的IL-27RαKO小鼠和WT对照组的能量消耗和营养消耗。正常饮食的IL-27RαKO小鼠的能量消耗在不改变食物摄入量的情况下显著减少(图7中a-c所示)。当喂食HFD时,表型更为显著(图1中a-c所示)。这些数据揭示IL-27信号具有维持全身代谢稳态的作用。为了确定IL-27Rα缺陷对能量消耗的影响是否可能归因于产热减少,本发明接下来检查了IL-27信号缺失对适应性热产生作用的影响。
IL-27RαKO小鼠在正常饮食条件下对冷诱导的低温敏感(图4中f和g所示),在HFD处理后,这种耐受不良更为严重(图1中d和e所示)。调控产热的关键蛋白解偶联蛋白1(UCP1)在IL-27RαKO小鼠中的表达也显著降低(图1中f-h所示)。组织切片也显示IL-27RαKO小鼠棕色脂肪组织(BAT)和皮下白色脂肪组织(SCW)中的褐变较少(图1中i所示)和含有多腔脂滴的细胞较少(图1中j所示)。这些数据清楚地表明IL-27RαKO小鼠的产热活性严重受损。值得注意的是,UCP1在IL-27RαKO小鼠中在冷刺激前已经降低(图1中f和h所示),与其他产热基因(图4中f所示)相同,这与***能量消耗减少一致(图1中b和c所示,图4中b和c所示)。此外,EBI-3KO小鼠的冷刺激重现了IL-27RαKO小鼠中的发现(图4中g-i所示),进一步证实了IL-27信号的致热效应。
接下来,解剖负责IL-27介导的信号传导的细胞类型。首先利用IL-27RαKO或WT小鼠的骨髓产生4组嵌合体小鼠(WT>WT、WT>KO、KO>WT和KO>KO),并对这些小鼠进行HFD处理。令人惊讶的是,只有那些用IL-27RαKO制备的嵌合体,无论供体细胞是什么,都极易受HFD诱导的脂肪积聚和冷刺激的影响(图2中a-c所示)。与野生型嵌合体相比,IL-27RαKO嵌合体的热发生始终受到抑制,表现为体温降低,多腔脂滴减少和UCP 1产生减少(图2中a-c所示)。这些研究表明IL-27的主导目标是脂肪细胞而不是免疫细胞。事实上,IL-27Rα在成熟脂肪细胞和原代米色脂肪细胞上表达。此外,IL-27Rα△adipo小鼠的热量产生也因对冷刺激的不耐受、褐变减少和UCP1产生减少而受到抑制(图2中c,e,f,h所示)。总之,上述数据表明IL-27通过直接靶向脂肪细胞来促进热量产生。
为了探讨IL-27对脂肪细胞作用的分子机制,我们在体外直接用IL-27处理原代米色脂肪细胞。IL-27处理后UCP1的表达显著增加,同时上调过氧化物酶体增殖物激活受体α(PPARα)、过氧化物酶体增殖物激活受体γ共激活剂1α(PGC-1α)和介导能量代谢的关键转录共激活物的表达(图3中a所示)。在免疫细胞中IL-27Rα的典型下游信号通路是JAK/STAT3和p38 MAPK。接下来我们检查IL-27是否通过这些途径促进脂肪细胞的热量产生。p38MAPK的磷酸化在IL-27刺激下显著增加,而pSTAT3仅短暂诱导并迅速消退(图3中b所示)。已知p38 MAPK激活ATF2介导的PGC-1α转录,并且PGC-1α是UCP1和热产生的主要调节者。正如预期的那样,IL-27处理也增强了原代米色脂肪细胞中ATF2的激活,并随之上调PGC-1α,其可通过药理学抑制p38 MAPK的活性来消除(图3中a-c所示)。此外,具有p38 MAPK抑制的细胞对IL-27诱导的UCP1促进耐受(图3中c所示)。然而,IL-27上调UCP1对STAT3抑制的反应基本上不受影响,尽管它也降低了PGC-1α的表达(图3中c所示)。这些结果表明,IL-27主要通过p38 MAPK-PGC-1α信号通路增强热量产生作用。有趣的是,p38和STAT3的抑制都不能消除PPARα的上调,提示IL-27的作用可能涉及额外的通路。
此外,向EBI-3KO小鼠补充IL-27可改善UCP1介导的热量产生并避免冷刺激中的低温(图4)。
总之,我们的结果表明IL-27信号是完整的适应性热量产生作用所必需的。IL-27直接靶向脂肪细胞诱导p38 MAPK的激活,从而驱动ATF2的激活和随后的PGC-1α和UCP1的表达。综上所述,这些发现提供了新的见解,扩大了我们对体内热量产生机理的理解。
尽管本发明已经参考示例性实施方案进行了描述,但应理解本发明不限于公开的示例性实施方案。在不背离本发明的范围或精神的情况下,可对本发明说明书的示例性实施方案做多种调整或变化。权利要求的范围应基于最宽的解释以涵盖所有修改和等同结构与功能。
Claims (13)
1.分子在制备用于控制生物体内热量产生的组合物中的用途,其特征在于,所述分子为用于调节EBI-3和p28结合的分子,或用于调节IL-27与其受体结合的分子。
2.根据权利要求1所述的用途,其特征在于,所述调节是指抑制或降低EBI-3和p28的结合,或者抑制或降低IL-27与其受体的结合,且所述控制是指抑制热量产生,进而降低所述生物体的体温。
3.根据权利要求2所述的用途,其特征在于,所述分子包括生物大分子或小分子化合物,其包括抗EBI-3抗体、抗p28抗体、抗IL-27R抗体、EBI-3可溶性片段、p28可溶性片段、IL-27R可溶性片段。
4.根据权利要求1所述的用途,其特征在于,所述分子为重组IL-27蛋白,所述控制是指促进热量产生,进而提高所述生物体的体温。
5.一种用于控制生物体热量产生的非治疗方法,其特征在于,向所述生物体施用用于调节EBI-3和p28结合的分子,或IL-27与其受体结合的分子。
6.根据权利要求5所述的方法,其特征在于,所述生物体包括人类受试者、哺乳动物或者微生物。
7.一种用于筛选能够控制生物体内热量产生的候选化合物的方法,其特征在于,包括测量施用待测化合物前后,EBI-3和p28结合情况,或IL-27与其受体的结合情况的步骤。
8.根据权利要求7所述的方法,其特征在于,当施用所述待测化合物后,EBI-3和p28的结合被抑制或降低、或者IL-27与其受体的结合被抑制或降低时,将所述待测化合物鉴定为抑制生物体内热量产生的候选化合物。
9.根据权利要求7所述的方法,其特征在于,当施用所述待测化合物后,EBI-3和p28的结合被促进或增强、或者IL-27与其受体的结合被促进或增强时,将所述待测化合物鉴定为促进生物体内热量产生的候选化合物。
10.根据权利要求7所述的方法,其特征在于,包括以下步骤:
(1) 测量生物体模型内EBI-3和p28结合的参数,或IL-27与其受体结合的参数,得到第一测量值的步骤;
(2) 向所述生物体模型施用待测化合物后,测量生物体模型内EBI-3和p28结合的参数,或IL-27与其受体结合的参数,得到第二测量值的步骤;和
(3) 比较第一测量值和第二测量值的步骤。
11.根据权利要求7所述的方法,其特征在于,包括以下步骤:
(1) 测量第一生物体模型内EBI-3和p28结合的参数,或IL-27与其受体结合的参数,得到第一测量值的步骤;
(2) 测量第二生物体模型内EBI-3和p28结合的参数,或IL-27与其受体结合的参数,得到第二测量值的步骤,其中所述第一生物体模型与所述第二生物体模型属于相同的模型,且所述第一生物体模型未施用待测化合物,所述第二生物体模型施用待测化合物;和
(3) 比较第一测量值和第二测量值的步骤。
12.根据权利要求7所述的方法,其特征在于,包括以下步骤:
(1) 测量施用待测化合物后生物体模型内EBI-3和p28结合的参数,或IL-27与其受体结合的参数,得到测量值的步骤;
(2) 将所述测量值与参考值比较的步骤,其中,所述参考值为由生物体模型在未施用待测化合物时测量得到的EBI-3和p28结合的参数,或IL-27与其受体结合的参数。
13.根据权利要求10-12任一项所述的方法,其特征在于,所述生物体模型包括动物模型或细胞模型。
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110986914.XA CN113694201B (zh) | 2021-08-26 | 2021-08-26 | 用于控制生物体内热量产生的组合物、方法及用途 |
PCT/CN2022/114928 WO2023025258A1 (zh) | 2021-08-26 | 2022-08-25 | 用于控制生物体内热量产生的组合物、方法及用途 |
EP22860604.2A EP4248998A4 (en) | 2021-08-26 | 2022-08-25 | COMPOSITION AND METHOD FOR REGULATING HEAT GENERATION WITHIN AN ORGANISM, AND USE |
KR1020247009619A KR20240052004A (ko) | 2021-08-26 | 2022-08-25 | 유기체 내 열량 발생을 제어하기 위한 조성물, 방법 및 용도 |
US18/321,062 US20230391843A1 (en) | 2021-08-26 | 2023-05-22 | Composition and method for controlling heat generation within organism, and use |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110986914.XA CN113694201B (zh) | 2021-08-26 | 2021-08-26 | 用于控制生物体内热量产生的组合物、方法及用途 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113694201A CN113694201A (zh) | 2021-11-26 |
CN113694201B true CN113694201B (zh) | 2022-09-02 |
Family
ID=78655048
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110986914.XA Active CN113694201B (zh) | 2021-08-26 | 2021-08-26 | 用于控制生物体内热量产生的组合物、方法及用途 |
Country Status (5)
Country | Link |
---|---|
US (1) | US20230391843A1 (zh) |
EP (1) | EP4248998A4 (zh) |
KR (1) | KR20240052004A (zh) |
CN (1) | CN113694201B (zh) |
WO (1) | WO2023025258A1 (zh) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113694201B (zh) * | 2021-08-26 | 2022-09-02 | 暨南大学 | 用于控制生物体内热量产生的组合物、方法及用途 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2018075740A1 (en) * | 2016-10-21 | 2018-04-26 | Merck Sharp & Dohme Corp. | Treating cancer with a combination of pd-1 antagonist and an il-27 antagonist |
Family Cites Families (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101039959A (zh) * | 2004-06-30 | 2007-09-19 | 杜门蒂斯有限公司 | 用于治疗炎性疾病的组合物和方法 |
CN101500610A (zh) * | 2006-08-11 | 2009-08-05 | 国立大学法人名古屋大学 | 抗肥胖药及其利用 |
WO2010118243A2 (en) * | 2009-04-08 | 2010-10-14 | Genentech, Inc. | Use of il-27 antagonists to treat lupus |
WO2011133931A1 (en) * | 2010-04-22 | 2011-10-27 | Genentech, Inc. | Use of il-27 antagonists for treating inflammatory bowel disease |
EP2700409A1 (en) * | 2012-08-23 | 2014-02-26 | Deutsches Rheuma-Forschungszentrum Berlin | IL-27 for modulation of immune response in acute lung disease |
JP2015047157A (ja) * | 2013-09-05 | 2015-03-16 | 国立大学法人佐賀大学 | 疼痛発症モデル動物および疼痛治療薬 |
WO2016073704A1 (en) * | 2014-11-06 | 2016-05-12 | Children's Research Institute, Children's National Medical Center | Immunotherapeutics for cancer and autoimmune diseases |
CN105854019A (zh) * | 2016-05-06 | 2016-08-17 | 暨南大学 | Il-27的受体激活剂在制备治疗肥胖症及其并发症产品中的应用 |
CN107661497A (zh) * | 2016-07-31 | 2018-02-06 | 复旦大学附属妇产科医院 | 白细胞介素(il)‑27及其受体抗体在制药中的用途 |
CN113694201B (zh) * | 2021-08-26 | 2022-09-02 | 暨南大学 | 用于控制生物体内热量产生的组合物、方法及用途 |
-
2021
- 2021-08-26 CN CN202110986914.XA patent/CN113694201B/zh active Active
-
2022
- 2022-08-25 EP EP22860604.2A patent/EP4248998A4/en active Pending
- 2022-08-25 WO PCT/CN2022/114928 patent/WO2023025258A1/zh unknown
- 2022-08-25 KR KR1020247009619A patent/KR20240052004A/ko active Search and Examination
-
2023
- 2023-05-22 US US18/321,062 patent/US20230391843A1/en active Pending
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2018075740A1 (en) * | 2016-10-21 | 2018-04-26 | Merck Sharp & Dohme Corp. | Treating cancer with a combination of pd-1 antagonist and an il-27 antagonist |
Non-Patent Citations (1)
Title |
---|
"Interleukin-27 decreases ghrelin production through signal transducer and activator of transcription 3—mechanistic target of rapamycin signaling";Sylvain Laverdure等;《Acta Pharm Sin B.》;20200107;第10卷(第5期);第838页右栏第2段 * |
Also Published As
Publication number | Publication date |
---|---|
EP4248998A1 (en) | 2023-09-27 |
CN113694201A (zh) | 2021-11-26 |
WO2023025258A1 (zh) | 2023-03-02 |
KR20240052004A (ko) | 2024-04-22 |
US20230391843A1 (en) | 2023-12-07 |
EP4248998A4 (en) | 2024-03-13 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Dmitrieva-Posocco et al. | Cell-type-specific responses to interleukin-1 control microbial invasion and tumor-elicited inflammation in colorectal cancer | |
Betz et al. | Batf coordinates multiple aspects of B and T cell function required for normal antibody responses | |
Boutant et al. | Mfn2 is critical for brown adipose tissue thermogenic function | |
Bageghni et al. | Cardiac fibroblast-specific p38α MAP kinase promotes cardiac hypertrophy via a putative paracrine interleukin-6 signaling mechanism | |
Meixner et al. | JunD regulates lymphocyte proliferation and T helper cell cytokine expression | |
Odegaard et al. | Alternative M2 activation of Kupffer cells by PPARδ ameliorates obesity-induced insulin resistance | |
Turpin et al. | Obesity-induced CerS6-dependent C16: 0 ceramide production promotes weight gain and glucose intolerance | |
Sinclair et al. | Lymphoid apoptosis and myeloid hyperplasia in CCAAT displacement protein mutant mice | |
Stiles et al. | Selective deletion of Pten in pancreatic β cells leads to increased islet mass and resistance to STZ-induced diabetes | |
Aouadi et al. | Inhibition of p38MAPK increases adipogenesis from embryonic to adult stages | |
Baldassarre et al. | Onset of natural killer cell lymphomas in transgenic mice carrying a truncated HMGI-C gene by the chronic stimulation of the IL-2 and IL-15 pathway | |
Ferretti et al. | Hypomorphic mutation of the TALE gene Prep1 (pKnox1) causes a major reduction of Pbx and Meis proteins and a pleiotropic embryonic phenotype | |
Rozas et al. | Targeted overexpression of tumor necrosis factor-α increases cyclin-dependent kinase 5 activity and TRPV1-dependent Ca2+ influx in trigeminal neurons | |
Hall et al. | Transforming growth factor-β3 (TGF-β3) knock-in ameliorates inflammation due to TGF-β1 deficiency while promoting glucose tolerance | |
Jansen et al. | Expression of the vanin gene family in normal and inflamed human skin: induction by proinflammatory cytokines | |
JP2009261956A (ja) | 造血及び血管生成の調節方法 | |
Zhang et al. | The LIM homeodomain transcription factor LHX6: a transcriptional repressor that interacts with pituitary homeobox 2 (PITX2) to regulate odontogenesis | |
Zirngibl et al. | Enhanced endotoxin sensitivity in fps/fes-null mice with minimal defects in hematopoietic homeostasis | |
Park et al. | A point mutation in the murine Hem1 gene reveals an essential role for Hematopoietic protein 1 in lymphopoiesis and innate immunity | |
CN113694201B (zh) | 用于控制生物体内热量产生的组合物、方法及用途 | |
Collier et al. | Pancreatic, but not myeloid-cell, expression of interleukin-1alpha is required for maintenance of insulin secretion and whole body glucose homeostasis | |
Zhang et al. | Nrf1 is time-dependently expressed and distributed in the distinct cell types after trauma to skeletal muscles in rats | |
Moenning et al. | Sustained platelet-derived growth factor receptor α signaling in osteoblasts results in craniosynostosis by overactivating the phospholipase C-γ pathway | |
US9109043B2 (en) | Screening method for antidiabetic agent using newly identified insulin secretion regulation factor | |
Li et al. | Enhanced skeletal muscle growth in myostatin-deficient transgenic pigs had improved glucose uptake in stretozotocin-induced diabetes |
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 | ||
TR01 | Transfer of patent right |
Effective date of registration: 20221207 Address after: Room 702, Block B, Cuiheng Building, No. 18 Hexin Road, Cuiheng New District, Zhongshan, Guangdong 528400 Patentee after: Guangdong Jiantbo Biotechnology Co.,Ltd. Address before: 510632 No. 601, Whampoa Avenue, Guangzhou, Guangdong Patentee before: Jinan University |
|
TR01 | Transfer of patent right |