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Objective: white female mice in this experiment replication UVB irradiation melasma experimental animal models given skin soup gavage, the final study of the biochemical indicators and pathological observations melasma mechanism of phase combination, to detect, analyze, and to further explore the pharmacodynamic mechanism of skin soup prevention chloasma; significant effect for clear skin soup treatment of melasma provide reliable experimental methods and theoretical basis. Materials and methods: select white female mice of clean grade 72, weight about 30g, screening qualified mice were randomly divided into six groups, each group of 12. Were blank control group, model control group, skincare the soup low dose group, skin soup in dose group, skincare the soup high dose group, the positive control group. White female mice UVB irradiation copy melasma experimental animal models. Group numbers, all the animals were housed separately. Before the experiment 24h all animal spine on both sides, and we rely on the buttocks the skin back hair removal with 10% NaS2 aqueous solution, respectively, then the residue with water to clean, to expose the back most of the skin. Weekly hair removal time, a total of four weeks before, drawing further one hair removal and drawn one hour before fasting. In addition to the normal control group, the wavelength of 320nm UVB irradiated mice daily skin on the back of irradiation time 60min, the animals and the distance from the light source to 20cm. The continuous irradiation around, and skin changes observed in mice hair removal site. The UVB irradiation white female mice 4 weeks after gavage skin decoction, in accordance with the standards of human clinical dose, for every 100 pounds of body weight given 285g skin Soup crude drug. Translation resulting average mice per 10g weight given 0.6g crude drug amount, per body weight of mice an average of 30 persons 2g only seen the average daily crude drug mice 0.18g / standard (equivalent to human clinical dose). Blank control group, model control group, two groups of animals were fed with normal saline 0.5ml / only for six weeks; skincare soup is low, medium and high dose groups were fed with 2, 4, and 8 times the human clinical standard dose skin soup decoction once a day for six weeks. The positive control group: given a vitamin C pill 0.1g / only, ground into powder, dissolved in 0.5ml of distilled water orally, once a day, for six weeks. Deadline drawn after 6 weeks of treatment, to remove the dead mice by gavage during, drawing a total of 72 mice in once again drawn hair removal, and in the last 9 drenching 60min after all mice were sacrificed, quickly whichever test liver and hair removal, skin tissue samples (1.5 * 1.5cm), rinsed after pre-cooling saline (4 ° C) specimens, and then drying with filter paper to remove blood and subcutaneous fat and connective tissue, and cut skin and liver specimens 0.5g, placed in a 9-fold, respectively, the tissue within the tissue specimen of a small tube of the precooling saline Shredded was poured into a glass homogenizer tube, with cell pulverizer homogenate (liver 2 skin four times), each 5S. The homogenate good 10% homogenate using a centrifuge to 3500r/min centrifugal 15min, the supernatant, in strict accordance with tyrosine SOD kit manual operation, the parameters were measured. Other were taken to the hair in mice skin a (1.0X 1.0cm), 10% formalin-fixed, paraffin-embedded sections, melanin markers using immunohistochemical techniques, dewaxing to water, antigen retrieval first antibody HMB45 (melanoma melanoma), two-step immunohistochemistry, DAB color, hematoxylin antibody was replaced with PBS as negative control for the detection of pathological morphology. Results: 1. Successful copy UVB irradiation white female mice chloasma experimental animal models. (1) UVB on mice skin, liver tyrosine the SOD content around the impact of ultraviolet radiation, compared to the model control group of mice with the blank control group, skin and liver tyrosine content was significantly higher significant differences (P lt; 0.05); skin and liver super oxide dismutase (SOD) activity was significantly decreased, with a significant difference (P lt; 0.05). (2) UVB on the number of melanocytes in the skin of mice, the number density, mean gray, with an average optical density content affect the model control group mice compared with the blank control group, the number of melanin-positive cells in the target for several density were significantly increased, with a significant difference (P lt; 0.05); decreased melanin-positive cells in the average gray, a significant difference (P lt; 0.05); melanin-positive cells in the mouse skin mean optical density was significantly increased, a statistically significance (P lt; 0.05). Skin soup can effectively affect indicators of melasma experimental animal models (1) skin soup on mouse skin, liver tyrosine SOD content of skin soup, in the low-dose group and model control group, tyrosine content decreased significantly, superoxide dismutase (SOD) activity was significantly higher, significant differences were statistically significant (P lt; 0.05), high skin Soup, middle dose group and positive control group In comparison, the tyrosine content was significantly reduced, superoxide dismutase (SOD) activity was significantly higher, the difference is significant, statistically significant (P lt; 0.05), skin soup low dose group compared with the positive control group, casein acid content and superoxide dismutase (SOD) activity without significant difference was not statistically significant (P gt; 0.05). (2) skin soup impact on the number of melanocytes in the skin of mice skin soup in the low-dose group after gavage skin soup, melanin-positive cell number downward trend compared with the model group, statistically significant (P lt; 0.05). Skin soup, middle dose group compared with the positive control group was significantly decreased, the difference was significant (P lt; 0.05). Skin soup low-dose group compared with the positive control group was not statistically significant (P gt; 0.05). (3) skin soup impact on the number of mouse skin density content high skin Soup, in the low-dose group after gavage skin soup, the number of melanocytes in the skin of mice a target density downward trend, with the model control group, statistically significant (P lt; 0.05). Skin soup, middle dose group compared with the positive control group was significantly decreased, the difference was significant (P lt; 0.05). Skin soup low-dose group compared with the positive control group was not statistically significant (P gt; 0.05). (4) skin soup melanin-positive cells of mouse skin mean gray skin the soup low, medium and high dose group fed with skin soup, melanin-positive cells in the skin of mice a target average gray upward trend compared with the model control group was statistically significant (P lt; 0.05). Skin soup, the high-dose group compared with the positive control group, improved significantly, the difference was significant (P lt; 0.05). Skin soup low-dose group compared with the positive control group was not statistically significant (P gt; 0.05). (5) skin soup impact on the mouse skin melanocytes average optical density skin soup is high in the low-dose group after gavage skin soup, mouse skin, a target melanin-positive cells mean optical density downward trend compared with the model control group was statistically significant (P lt; 0.05). Skin soup, middle dose group compared with the positive control group was significantly decreased, the difference was significant (P lt; 0.05). Skin soup low-dose group compared with the positive control group was not statistically significant (P gt; 0.05). Conclusion: Copy UVB irradiation white female mice the melasma experimental animal model success. Skin soup prevention chloasma mechanism antioxidant, reducing the tyrosine content of the skin and liver tissues, increase SOD activity, effectively inhibit skin melanocyte proliferation, thereby reducing melanin synthesis related. The 3 skin Decoction chloasma significant.
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