异鼠李素抑制H_(2)O_(2)诱导的黑素细胞ROS水平升高和细胞活性降低

Isorhamnetin inhibits H_(2)O_(2)-induced ROS level increase and cell activity decrease in melanocytes

目的探讨异鼠李素(isorhamnetin,ISO)对人表皮黑素细胞免受H_(2)O_(2)诱导氧化应激损伤的预保护作用。方法不同浓度的H_(2)O_(2)处理人表皮黑素细胞,CCK-8检测细胞活性,筛选出H_(2)O_(2)体外诱导人黑素细胞氧化应激的最适浓度;用ISO预处理细胞后,使用最适浓度H_(2)O_(2)刺激,CCK-8检测细胞活性,NaOH裂解法测定黑素含量,流式细胞术检测细胞内ROS水平。结果0.8mmol/L H_(2)O_(2)处理人表皮黑素细胞24h后细胞活性显著降低;ISO预处理可使人黑素细胞活性显著增强,黑素合成增加,并可使H_(2)O_(2)刺激细胞内ROS水平明显降低。结论ISO对H_(2)O_(2)引起的正常人表皮黑素细胞的氧化损伤具有保护作用。该结果为抗氧化剂ISO应用于白癜风的治疗提供了一定的实验基础。

Objective To investigate the protective effect of isorhamnetin(ISO)on human melanocytes against H_(2)O_(2)-induced oxidative stress.Methods Human epidermal melanocytes were treated with different concentrations of H_(2)O_(2).The cell activity was detected by CCK-8 to screen the optimal concentration of H_(2)O_(2) to induce oxidative stress in human melanocytes in vitro.After ISO pretreatment,the cells were stimulated with the optimal concentration of H_(2)O_(2).The cell activity was detected by CCK-8,the melanin content was measured by the NaOH lysis method,and the intracellular ROS level was detected by flow cytometry.Results After being treated with 0.8mmol/L H_(2)O_(2) for 24h,the cell activity of human epidermal melanocytes decreased significantly.ISO pretreatment could significantly enhance the cell activity and melanin synthesis in human melanocytes and significantly reduce the H_(2)O_(2)-induced ROS.Conclusion ISO has a protective effect on human epidermal melanocytes against the oxidative damage induced by H_(2)O_(2).This finding provides a specific experimental basis for the application of antioxidant ISO in the treatment of vitiligo.