CN103735566A - Application of soracan gum in inhibiting skin injury of ultraviolet ray - Google Patents
Application of soracan gum in inhibiting skin injury of ultraviolet ray Download PDFInfo
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- CN103735566A CN103735566A CN201310730754.8A CN201310730754A CN103735566A CN 103735566 A CN103735566 A CN 103735566A CN 201310730754 A CN201310730754 A CN 201310730754A CN 103735566 A CN103735566 A CN 103735566A
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Abstract
The invention discloses application of soracan gum in inhibiting skin injury of ultraviolet ray, and relates to application of soracan gum with different mass concentrations in protecting skin from injury due to ultraviolet ray UVA. A cell model adopted in the invention is an ultraviolet ray UVA induced cell injury model, wherein the dosage of soracan gum is 0.625-5 mu g/mu l; a mouse model adopted in the invention is an ultraviolet ray UVA induced mouse skin injury model, and the dosage of the soracan gum is 200mu l of 1 percent solution, the soracan gum can be used for improving the cell survival rate of cells in ultraviolet radiation, reducing inflammation factors in mouse skin and improving gene expression of synthetic collagen, so that skin injury caused by ultraviolet ray UVA can be inhibited. Cell and animal experiments prove that soracan gum has a remarkable effect of inhibiting skin injury of ultraviolet ray, so that soracan gum can be used in processes of preventing and repairing skin injury due to ultraviolet ray UVA, and can be used for preparing anti-ultraviolet radiation injury medicines and anti-ultraviolet radiation cosmetics.
Description
Technical field
The invention belongs to medicine, cosmetics technical field, the present invention relates to a kind of Salecan and suppressing ultraviolet to the application in skin injury.
Background technology
Ultraviolet is the general name of electromagnetic spectrum medium wavelength from 10nm to 400nm radiation, according to the difference of ultraviolet self wavelength, mainly ultraviolet is divided into three regions, i.e. short wave ultraviolet (UVC), ultraviolet B radiation (UVB) and long wave ultraviolet (UVA).Ultraviolet is different to the penetration degree of human body skin.Long wave ultraviolet can reach deep dermis, causes that dermal melanin is calm, makes skin tanning, lipid and collagen protein impaired, and accumulation, is the main cause that causes skin aging and grievous injury for a long time.
At present, the anti-ultraviolet sunscreen product on market mainly contains the sunscreen products that physical property sunscreen products, chemical sunscreen products and physical chemistry combine.What physical sunscreen frost was conventional is some very thin mineral particles; as zinc oxide, titanium dioxide etc.; also can add some Pulvis Talci and calcium simultaneously; form layer protecting film; be used for scattering and absorb ultraviolet, its shortcoming is: must be coated with very thickly, more greasy; easily stop up pore, and in the sun due to perspiration dissolve affect attractive in appearance.Chemistry is sun-proof is to utilize chemical reaction, for skin, absorb the ultraviolet in sunlight, rear effect nature for chemical sunscreen, seem relatively slim and graceful thoroughly clear, this is the advantage of chemical sun-proof maximum, and shortcoming is: chemical sunlight screening additive is more, and some effective ingredient wherein may cause the allergy of indivedual skin, and after being coated with, just play a role half an hour, and life-time service has certain injury to skin.The sunscreen products that Chemical Physics combines is the advantage that has merged chemical and two kinds of sunscreen productses of physical property, use commonplace.
Last century the forties, doctor Diluzio of Tu Lun university first from yeast cell wall separation obtain beta glucan, beta glucan has just obtained widely paying close attention to.In recent years, beta glucan has been widely used in the fields such as medicine, food, personal care articles, feedstuff, as the functional cosmetics that contains beta glucan can nourishes rough skin, improve skin appearance, and various dermatosiss are had to curative effect (Singh RP energetically, Agarwal R.Cosmeceuticals and silibinin.Clin Dermatol, 2009,27:479-484.).Beta glucan is a kind of natural high polysaccharide, can on skin, form layer of transparent, flexible, ventilative thin film, the effectively infringement of harmful substance to skin in isolated environment.
Salecan is the extracellular polysaccharide that a kind of edaphic bacillus Agrobacterium sp.ZX09 secretion produces, and structure is beta glucan (Chinese patent, 201010146371.2).Through the evaluation of emergency toxicology and chronotoxicology, Salecan non-toxic and safe.
Found that at present Salecan has reduction blood glucose (Chinese patent, 201010146371.2) and reduce liver fat, blood fat, body fat, body weight and cholesterol (Chinese patent, 201010198223.5) and prevention and treatment constipation effect, the present invention is for to find that the skin injury that Salecan causes ultraviolet (UV) A has certain inhibition and/or repair first.
Summary of the invention
The object of the invention is to provide a kind of Salecan suppressing ultraviolet to the application in skin injury, that this Salecan has is easy to use, suppress and/or repair the function of ultraviolet to the damage of skin.
The technical scheme that realizes the object of the invention is: a kind of Salecan, suppressing ultraviolet to the application in skin injury, is directly smeared Salecan solution and skin surface.
The mass concentration of described Salecan solution is 0.5-1.5%.
Described ultraviolet is that ultraviolet (UV) A causes to the damage of skin.
Described application comprises people's mammal.
Compared with prior art, the present invention has the following advantages:
1. the production of the Salecan in the inventive method, simple and convenient extraction, can directly be dissolved in cold water, and its solution has excellent anti-ultraviolet function, has increased skin to ultraviolet toleration.
2. Salecan solution can improve the survival rate of skin flbroblast under ultraviolet (UV) A, in zoopery, with the normal mouse ratio irradiating through ultraviolet (UV) A, the inflammatory cytokine of smearing in the mouse skin tissue of Salecan solution obviously reduces, the related gene of synthetic collagen protein obviously improves, and HE section shows that Salecan has obviously alleviated the damage of ultraviolet (UV) A to skin.
3. after Salecan wiring solution-forming, can directly be applied in mammal skin, Salecan solution forms one deck protective film at skin surface, can effectively keep out ultraviolet, can be used for preparation and has the functional product of inhibition ultraviolet to the sun-proof series of skin injury.
Accompanying drawing explanation
Fig. 1 is that Salecan improves the cell survival rate of NIH3T3 l cell after ultra-vioket radiation.
Fig. 2 be Salecan on ultra-vioket radiation after the impact (A is the mrna expression of TNF-α, and B is the mrna expression of IL-1 β) of mouse skin expression of inflammatory cytokines.
Fig. 3 be Salecan on ultra-vioket radiation after the impact (A is the mrna expression of Col1 α 1, and B is the mrna expression of Col3 α 1) expressed of mouse skin collagen gene.
Fig. 4 be Salecan on ultra-vioket radiation after the impact of mouse skin metamorphosis.
The specific embodiment
Below in conjunction with embodiment, the present invention is described in further detail.
The preparation of embodiment 1 Salecan solution
The preparation of Salecan solution comprises the following steps:
1) Salecan crude product adopts the Sevag method purification after improving.Purification step is:
A. prepare 0.5% Salecan solution, (105 ℃, 15min), to be cooled during to room temperature, 7000r/10min, gets supernatant, to remove remaining thalline in heating.
B. in the ratio of 1:30, add the acetic acid of pH4.0---sodium acetate solution, to remove DNA.
C. in the ratio of 1:1, supernatant is mixed with phenol chloroformic solution (ratio in 1:1 is mixed with mixed solution), the about 15min of thermal agitation, 7000r/10min, gets supernatant.Repeat this step 2～3 time, thoroughly to remove Deproteinization.
D. to the industrial alcohol that adds 2 times of volumes in supernatant, sugar flocculates into bulk, pulls out, squeezes and removes ethanol.
E. use 75% ethanol washing precipitation 2～3 times, then use 95% ethanol washing precipitation 1～2 time.
F. vacuum lyophilization.
2) take the 30mg Salecan obtaining through step 1), be dissolved in 30ml distilled water, until completely dissolved, add 6 ‰ phenoxyethanol (antiseptic), after mix homogeneously, carry out being packaged as independent bottle after conventional sterilizing, be 1wt% Salecan solution.For the Salecan solution of cell experiment, without adding antiseptic, and to guarantee sterile working during subpackage.
Experimental cell is NIH3T3 l cell.Under standard test conditions, cultivate: cell is used containing the DMEM high glucose medium of the mycillin of 10% calf serum and 1% and cultivates (complete medium), is placed in 37 ℃, 5%CO
2biochemical cultivation case in, within every 2～3 days, change a subculture.Cell is used the trypsin solution containing 0.25% pancreas enzyme-EDTA to digest, go down to posterity.
Experiment adopts to be carried out after different disposal cell, and ultraviolet (UV) A irradiates and causes cell injury, and after irradiating by detection ultraviolet (UV) A, the survival rate of cell, evaluates cell injury degree and the degree of protection of Salecan to cell.
NIH3T3 l cell is with 2 * 10
4density kind, in 96 orifice plates, arranges and is divided into 6 groups, every group of 5 holes.Experiment grouping situation is: CK, and UVA, Salecan component is 0.625 μ g/ μ l, 1.25 μ g/ μ l, 2.5 μ g/ μ l, tetra-dosage groups of 5 μ g/ μ l, wherein CK is experiment contrast group, UVA group and tetra-dosage groups of Salecan.Cultivate after 24 hours, discard culture medium, every Kong Junxian add 100 μ l not containing the DMEM culture medium of calf serum, by group, process, wherein CK group and UVA organize and respectively add 100 μ lPBS again, and tetra-dosage groups of Salecan respectively add 0.125%, 0.25%, 0.5% and 1% the Salecan solution of 100 μ l, putting into incubator cultivates, after 3 hours, the ultraviolet (UV) A that UVA group and Salecan group are accepted 30min irradiates, and radiation dose is 18J/cm
2, radiation length is 20cm.After irradiation finishes, together with CK group, discard together culture medium, be replaced by complete medium.After 24 hours, by triumphant base MTT cell proliferation/toxicity detection kit, carry out cell survival rate detection.
Embodiment 3 Salecans to ultra-vioket radiation after the research of mouse skin model
Laboratory animal is the male C 57 BL/6 J mouse in 8 week age.Under standard test condition, raise: 12 hours illumination-12 hour dark cycle, freely absorb water and food.By all experiment mices, by lumbar injection 10% chloral hydrate (4ml/kg) anesthesia, wait anaesthetizing when abundant, fixedly mice makes its back upward.Take back vertebra as intermediate line, with shears, back Mus hair is cut off, size is 2 * 2cm approximately.After one week, mice is divided into 3 groups at random, is divided into CK group, UVA group, Salecan group, and wherein, CK group is Normal group, and UVA group is ultraviolet (UV) A irradiation group, and these two groups give corresponding solvent (being distilled water) and process in experimentation; Salecan group, for Salecan solution-treated group, is being accepted ultraviolet (UV) A pre-irradiation, smears the Salecan solution of 200 μ l, accepts ultra-vioket radiation after half an hour, within every 4 hours, smears skin surface once.UVA group and Salecan group are accepted the ultraviolet (UV) A irradiation of 48h altogether in 6 days, and radiation dose is 18J/cm
2, radiation length is 20cm.Last irradiation after 24 hours, puts to death mice, gets the skin histology in unhairing region, back, for evaluating skin degree of impairment.
1) extraction of skin histology RNA
Get fresh skin of back and organize about 25mg, according to Invitrogen RNA, extract the explanation of test kit and M-MLV reverse transcription test kit, extract RNA and reverse transcription cDNA.After cDNA stock solution is diluted 20 times, for RT-PCR reaction, reaction system is 20 μ l:
Pcr amplification reaction condition is: 50 ℃, and 2min; 95 ℃, 10min; 95 ℃ of 15s, 60 ℃ of 30s, 40 circulations; 95
Gene relative expression quantity is with 2
-△ △ CTmethods analyst, take GAPDH as internal reference.
3) expression of inflammatory cytokine in skin histology
Ultra-vioket radiation can cause that inflammatory reaction appears in skin, secretion produces a large amount of inflammatory cytokine, wherein, tumor necrosis factor (TNF-α) and il-1 β (IL-1 β) are two kinds of Main Factors of proinflammatory phase, and the abnormal rising of these inflammatory cytokine makes skin occur the phenomenons such as red and swollen.As shown in Figure 2, in UVA group, the expression of TNF-α and IL-1 β is obviously risen compared with CK group, and in Salecan group, the expression of TNF-α recovers normally substantially, and the expression of IL-1 β obviously reduces.Result shows, Salecan can obviously reduce the expression of inflammatory cytokine, alleviates the inflammatory reaction of skin after ultra-vioket radiation.
4) expression of collagen gene in skin histology
Ultra-vioket radiation causes the destruction of collagen fibers of skin, reduces the content of collagen protein in skin, makes cutis laxa, occurs the catabiosis such as microgroove.Col1 α 1 and Col3 α 1 are respectively the main synthetic gene of NTx and III Collagen Type VI.As shown in Figure 3, compare with CK group, the Col1 α 1 of UVA group and the gene expression of Col3 α 1 reduce, and the Col1 α 1 of Salean group and the gene expression of Col3 α 1 and CK group are basic identical.Result shows, Salecan can obviously be protected the collagen protein of skin, reduces the injury of ultraviolet (UV) A to skin.
3) skin histology Epidemiological Analysis
Skin histology is fixed with 10% formalin, specimens paraffin embedding slices then, then dye with hematoxylin-eosin HE.By the degree of impairment of microscopic examination skin.As shown in Figure 4, CK group epiderm skin thickness homogeneous, skin corium arrangement of collagen fibers is neat; UVA group skin corium thickens, and arrangement of collagen fibers is disorderly, obviously degeneration, skewness; Salecan group epidermal thickness approaches normal, and arrangement of collagen fibers is evenly neat, compared with UVA group, has clear improvement, and approaches CK group.
Claims (4)
1. Salecan is suppressing ultraviolet to the application in skin injury.
2. application according to claim 1, is characterized in that described ultraviolet is that ultraviolet (UV) A causes to the damage of skin.
3. application according to claim 1, is characterized in that described Salecan is that mass concentration is the solution of 0.5-1.5%.
4. application according to claim 1, is characterized in that described application comprises people's mammal.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111773239A (en) * | 2020-07-31 | 2020-10-16 | 四川合泰新光生物科技有限公司 | Application of beta-1, 3-glucan in preparation of medicines and daily chemical products for repairing skin injuries |
| CN111778300A (en) * | 2020-07-31 | 2020-10-16 | 四川合泰新光生物科技有限公司 | Beta-1, 3-glucan and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE19710368A1 (en) * | 1997-03-13 | 1998-09-17 | Henkel Kgaa | Use of water-soluble beta-glucans as active ingredients for the production of therapeutic agents for skin treatment |
| WO2000038645A1 (en) * | 1998-12-28 | 2000-07-06 | Celanese Ventures Gmbh | Utilisation of water-insoluble linear poly-alpha-glucan as uv filter |
| CN1347306A (en) * | 1999-03-12 | 2002-05-01 | 生物技术公司 | Sun protection agents |
-
2013
- 2013-12-26 CN CN201310730754.8A patent/CN103735566A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE19710368A1 (en) * | 1997-03-13 | 1998-09-17 | Henkel Kgaa | Use of water-soluble beta-glucans as active ingredients for the production of therapeutic agents for skin treatment |
| WO2000038645A1 (en) * | 1998-12-28 | 2000-07-06 | Celanese Ventures Gmbh | Utilisation of water-insoluble linear poly-alpha-glucan as uv filter |
| CN1347306A (en) * | 1999-03-12 | 2002-05-01 | 生物技术公司 | Sun protection agents |
Non-Patent Citations (1)
| Title |
|---|
| 鲁晋南: "《索拉胶（Soracan gum）的流变学性质及其生物活性》", 《中国优秀硕士学位论文全文数据库》, 1 June 2008 (2008-06-01) * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111773239A (en) * | 2020-07-31 | 2020-10-16 | 四川合泰新光生物科技有限公司 | Application of beta-1, 3-glucan in preparation of medicines and daily chemical products for repairing skin injuries |
| CN111778300A (en) * | 2020-07-31 | 2020-10-16 | 四川合泰新光生物科技有限公司 | Beta-1, 3-glucan and preparation method and application thereof |
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Application publication date: 20140423 |