【摘要】 目的建立组织培养人皮肤模型并探讨驱虫斑鸠菊提取物[Vernonia anthelmintica(L.)extract,AVE]在该模型下对人皮肤组织黑色素合成功能的影响及可能的作用机制。方法采用HE染色法观察不同培养条件对组织培养的人皮肤形态的影响,HE染色法和乳酸脱氢酶(LDH)释放法检测不同浓度AVE对人皮肤组织毒性的影响,NaOH裂解法、L-Dopa氧化法检测AVE对人皮肤组织黑色素含量及酪氨酸酶活力的影响,免疫组化法和Western blot法观察AVE对组织培养的人皮肤S100和黑色素合成关键蛋白表达的影响。结果以William’s E培养基为基础培养的人皮肤组织结构完整,形态良好。在组织培养的人皮肤模型中,AVE剂量依赖性的促进黑色素合成和酪氨酸酶活性。AVE能够上调黑色素合成关键调控因子MITF及相关蛋白(TYR、TRP-1和TRP-2)与S100的表达。结论 AVE通过上调MITF及其调控的黑色素合成相关蛋白表达与黑色素细胞的增殖促进人皮肤组织的黑色素生成。更多还原

【Abstract】 Objective To establish the human skin organ culture model and to investigate the effect of Vernonia anthelmintica(L.) extract(AVE) on the melanogenesis of human skin and its potential mechanism. Methods H&E staining was performed to screen the culture conditions for human skin. H&E staining and lactate dehydrogenase(LDH) activity were used to observe the toxicity on organ culture of human skin under different concentrations of AVE. Na OH dissolution and L-Dopa oxidation were used to detect the melanin content and tyrosinase activity. The melanogenic proteins(TYR, TRP-1, TRP-2 and MITF) and S100 were analyzed by Western blot and immunohistochemical method. Results The skin structure remained complete and integrated after organ culture with William’s E medium. Different concentrations of AVE had no toxicity on the organ cultured human skin; AVE increased the melanin content and tyrosinase activity in a dose-dependent manner. AVE up-regulated the expression of S100 and melanogenic proteins(TYR, TRP-1, TRP-2 and MITF). Conclusion AVE may promote melanogesnesis in organ cultured hum an skin by increasing the tyrosinase activity, stimulating melanocyte proliferation and activating the expression of melanogenic proteins.更多还原