| Purpose:This experiment copy medium-wave ultraviolet against white female mice to build mode method, skin, will eventually serve soup filling the biochemical research income and pathology observations and age spot mechanism combined, inspection, analysis, further to explore the pharmacodynamics yellow-brown spots skin soup prevention mechanisms; To ascertain the age spot skin decoction "in the treatment of significant effect offer reliable experimental methods and theoryMaterial and method:Select clean level white female mice, weight around 30g 72 only mice, will screening qualified at random into six groups, each group of 12 only. Separately for blank control; Model control; Low dose group; skin soup Dose group of beautiful skin soup; Beautiful skin soup high-dosage groups; Positive drug control group. Using ultraviolet irradiation white female mice characterist.the experimental animal model. Copy yellow-brown spots Press group Numbers, all animals points cage raising. In experiments 24h before all animals on both sides and spine skin with 10% respectively NaS2 hip removal, then use solution back water residue clean, to expose back most of the skin. Once a week, were depilate, based around before another hair removal, and in an hour before alternately fast. In addition to the normal control group in the 320nm mw ultraviolet wavelength for daily illuminate skin 1, the mice back 60min irradiation time, animals and light source for the distance 20cm. Continuous radiation around, and observe the depilation parts skin changes in mice. Using ultraviolet irradiation white female mice characteristics.the 4 weeks, irrigation, beautiful skin soup decoction served in accordance with human clinical dosage standards, body weight per 100 jins of 285g skin beautifying soup born to dose. Conversion income, average mice every 10g weight give 0.6 g per mice born dosage, the average 30 and weight, predictably 2g dose daily average mice born for 0.18g/only. (equivalent to human clinical standard doses). Normal controls, two groups of animals were model respectively take physiological saline irrigation, the bid only 0.5 ml/, and lasts for six weeks; Beautiful skin soup low, medium and high doses groups were irrigation clothing 2,4,8times human clinical standard doses, the bid skin soup decoction for six weeks. Positive drug control: to give vitamin C pills 0.1 g/only, grinding into powder, soluble in 0.5ml of distilled water irrigation, the bid, continuous served six weeks. Treatment six weeks, drawn, remove irrigation deadline of mice, deceased during the stomach of 72 only in mice alternately drawn once more before removal, and in the last irrigation 60min all mice after drug broken neck executed, quickly take its biooxyrate liver and hair removal skin tissue samples (1.5*1.5cm),4°C, via (gets cold saline), remove the blood and rinse specimens hypodermic and adipose and other connective tissue, then with the filter, and slice the drying skin and liver specimens taken each 0.5g, nine times respectively in tissue samples in advance of the cold saline bench tube, will organize shear ground into glass homogenate tube, the shredder homogenate (with liver cell 2 times, four times), skin every 5S. Will homogenate good 10% evenly with centrifuge 3500r/slurry with 15min, take min centrifugal and strictly supernatant fluid, SOD kit specifications of tyrosinase, index determination. In addition to hair skin mice were taken blocks (1.0X1cm), 10% 1.0 formalin fixed, conventional paraffin embedding slice, melanin marker adopts immunhistochemical techniques, then after dewaxing to water, the antigen, and HMB45 (repaired, with melanoma) for the first anti body,immunohistochemical two footwork, DAB show color and hematoxylin-eosin grain after dyeing. Instead of a resistance to do with PBS negative control. Used for pathological morphology detection.。Results:1.copy ultraviolet irradiation white female mice mw of experimental animals model success yellow-brown spots.(1) UVB on mice in the skin, liver, SOD the influence of tyrosine content Medium-wave ultraviolet radiation within four weeks, model control group, compared with that of normal mice skin and liver was obviously increase, with tyrosine significant difference(P<0.05). The skin and super oxide dismutase (liver SOD activity reduced significantly, and has significant difference(P <0.05).(2) UVB to the number of the skin cancer cells in mice, number density, average gray, average light density content influencesModel control group group compared with normal mice, target the number of positive cells within the melanin were significantly increased, numerical density with significant difference(P<0.05). Melanin positive cells average gray declined obviously, and significant difference (P<0.05). The skin melanin positive cells in mice average light density of increasing, the content was statistical significance (P<0.05).2.Beautiful skin soup can effectively influence age spot index of animal experiment model(1) skin beautifying skin, liver of mice soup in the influence of tyrosine, SOD contentBeautiful skin soup high, medium and low dose group, compared with model control group was obviously lower tyrosine, superoxide dismutase (SOD) activity increased significantly, was statistically significant difference (P<0.05), skin beautifying soup with high, middle dose group of positive drug rcpmaj tyrosine content decreased remarkably, are superoxide dismutase (SOD) activity increased significantly, was statistically significant difference (P<0.05), skin beautifying soup with low dose group, positive drug rcpmaj tyrosine content and superoxide dismutase (SOD) activity no significant differences, no statistically significant (P>0.05).(2) skin beautifying skin melanin cell in mice soup decoction of the number, number density, average gray, average light density content influences Beautiful skin soup high, medium and low dose group served by skin soup, filling the number positive cells melanin declined, compared with model control group was statistically significant (P<0.05). Beautiful skin soup with high, middle dose group of positive drug rcpmaj, declined obviously, and significant difference (P<0.05). Low dose group and beautiful skin soup positive drug rcpmaj, no statistically significant (P>0.05).(3) to human skin skin soup number density content influencesBeautiful skin soup high, medium and low dose group served by skin soup, filling in mice within skin target declined, number density with model control group was statistically significant comparative, (P<0.05). Beautiful skin soup with high, middle dose group of positive drug rcpmaj, declined obviously, and significant difference (P<0.05). Low dose group and beautiful skin soup positive drug rcpmaj, no statistically significant(P>0.05).(4) skin beautifying soup decoction of the skin melanin positive cells in mice average light density effectBeautiful skin soup low, medium and high dose group of beautiful skin soup by filling after served in mice within skin target mean grey positive cells melanin an upward trend with model control group, was statistically significant comparative,(P<0.05). Beautiful skin soup, high dose group and positive drug rcpmaj, improved obviously and significant difference (P<0.05). Low dose group and beautiful skin soup positive drug rcpmaj, no statistically significant(P>0.05).(5) skin beautifying skin melanin cell in mice soup decoction of the influence of the average optical densityBeautiful skin soup high, medium and low dose group served by skin soup, filling in mice within skin target average light density melanin positive cells declined, with model control group was statistically significant comparative, (P<0.05).Beautiful skin soup with high, middle dose group of positive drug rcpmaj, declined obviously, and significant difference(P<0.05). Low dose group and beautiful skin soup positive drug rcpmaj, no statistically significant (P >0.05).Conclusion:1. Copying medium-wave ultraviolet irradiation white female mice age spot animal experiment model made mould success.2. The mechanism of prevention and cure skin soup yellow-brown spots may be and antioxidant, and effectively suppress melanocytes proliferation, reduce tyrosinase activity, which reduces dark meat synthetic relevant.3. Beautiful skin decoction age spot curative effect is distinct. |
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