芍药苷对黑素细胞氧化损伤分子机制的研究

Molecular Mechanism of Regulating Melanocytes Oxidative Damage by Paeoniflorin

目的观察芍药苷各组抑制B16黑素细胞氧化损伤的分子机制。方法体外细胞实验分为对照组、模型组、不同浓度芍药苷干预组。采用流式细胞仪检测细胞凋亡率,Hoechst 33342观察凋亡细胞的核变化,激光共聚焦显微镜观察细胞线粒体膜电位,底物荧光法检测Caspase-3/9的活性,Western blot法测定细胞凋亡Bcl-2、Cyt-c蛋白的表达。结果(1)与正常对照组相比,模型组凋亡率明显增高(P<0.01);Caspase-3/9的活性显著增高;Cyt-c蛋白表达量升高;Bcl-2蛋白表达量降低。(2)与模型组相比,芍药苷各组凋亡率明显降低(P<0.01);Caspase-3/9的活性降低,Cyt-c蛋白表达量降低,Bcl-2蛋白表达量升高,均以芍药苷高剂量组最好。结论该文揭示了线粒体信号转导通路可能是芍药苷干预氧化应激诱导B16黑素细胞凋亡的调控机制之一。

Objective To investigate the molecular mechanism of paeoniflorin to inhibit the oxidative damage of B16 melanocytes.Methods In vitro experiments cells were divided into the control group,the model group and different concentrations of paeoniflorin intervention groups.The apoptosis rate was detected by flow cytometry.Hoechst 33342 was used to observe the nuclear changes of apoptotic cells.The mitochondrial membrane potential was observed by laser confocal microscope,the activity of Caspase-3/9 was detected by substrate fluorescence assay and the expressions of Bcl-2 and Cyt-c protein in apoptosis cells were determined by Western blot.Results(1)Compared with that of the control group,the apoptosis rate in the model group was significantly increased(P<0.01).The mitochondrial membrane potential was decreased.The activity of caspase-3/9 was significantly increased.The expression of Cyt-C protein was increased and the expression of Bcl-2 protein was decreased.(2)Compared with that of the model group,the apoptosis rates of paeoniflorin intervention groups were significantly lower(P<0.01).The activity of Caspase-3/9 was decreased.The expression of Cyt-C protein was decreased and the expression of Bcl-2 protein was increased,all of which were best in the high-dose paeoniflorin group.Conclusion This paper reveals that mitochondrial signal transduction pathway may be one of the regulatory mechanisms of paeoniflorin intervention in B16 melanocyte apoptosis induced by oxidative stress.