地西泮保护黑色素细胞氧化损伤的作用及机制

Protective effect and mechanism of diazepam on oxidative damage of melanocytes

构建了小鼠黑色素细胞B16F10的氧化应激模型,研究了地西泮对B16F10细胞氧化损伤的保护作用及相关机制。选取生长状态良好的B16F10细胞,采用细胞活力实验(MTT)确定氧化应激造模的双氧水浓度和地西泮的合适剂量范围,通过荧光检测验证地西泮对B16F10细胞氧化损伤的保护作用,采用免疫蛋白印迹方法(Western blot,WB)进一步探究地西泮产生保护作用的上游通路机制。实验结果发现,地西泮能够显著逆转细胞的氧化损伤,恢复细胞活力,升高应激状态下抗凋亡蛋白B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)的表达,降低促凋亡蛋白Bax(Bcl2-Associated X)的水平,最终保护氧化应激状态下的B16F10细胞。

The oxidative stress model of mouse melanocytes B16F10 was established and the protective effect and mechanism of diazepam on oxidative damage of B16F10 cells were studied.B16F10 cells in good growth state were selected to determine the concentration of hydrogen peroxide and the appropriate dose range of diazepam by MTT method.The protective effect of diazepam on oxidative damage of B16F10 cells was observed by fluorescence detection.Western blot was used to further explore the upstream pathway mechanism of diazepam's protective effect.The experimental results showed that diazepam can significantly reverse cellular oxidative damage,restore cell vitality,increase the expression of anti apoptotic protein Bcl-2 under stress,reduce the level of pro apoptotic protein Bax,and ultimately protect B16F10 cells under oxidative stress.