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目的:探讨黄芩苷对鱼藤酮致PC12细胞损伤的保护作用及其机制。方法:建立鱼藤酮损伤PC12细胞模型,采用终浓度分别为0.1,1,10μmol·L-1的黄芩苷进行药物干预;四氮唑盐(MTT)法检测细胞活力,并用显微镜观察细胞形态;应用Annexin V-FITC/PI双染流式细胞术检测细胞凋亡;采用荧光探针二氯荧光素-双乙酸盐(DCF-DA)测定细胞内氧自由基(ROS);采用蛋白免疫印记法检测凋亡信号蛋白Bcl-2,Bax及Caspase-3表达。结果:PC12细胞经1.5μmol·L-1鱼藤酮孵育24 h后细胞活力显著降低,黄芩苷对鱼藤酮所致的细胞损伤呈现剂量依赖性的保护作用;流式细胞术分析表明黄芩苷可明显减少鱼藤酮致PC12细胞凋亡比例;荧光显微镜观察发现黄芩苷能明显减少鱼藤酮损伤PC12细胞ROS含量,免疫印迹实验表明黄芩苷可上调模型细胞Bcl-2及下调Bax表达并减少活化Caspase-3含量。结论:黄芩苷对鱼藤酮诱导的PC12损伤具有保护作用活性,其机制可能与黄芩苷减少鱼藤酮诱导PC12细胞ROS含量,抑制线粒体凋亡通路有关。
Objective: To investigate the protective effect of baicalin on rotenone-induced PC12 cell injury and its mechanism. Methods: The rotenone-induced PC12 cell model was established, and the drug intervention was induced by baicalin at final concentration of 0.1, 1 and 10μmol·L-1 respectively. The cell viability was detected by MTT assay and the cell morphology was observed by microscope. Annexin Cell apoptosis was detected by V-FITC / PI double staining flow cytometry. Intracellular oxygen free radicals (ROS) were detected by fluorescent probe dichlorofluorescein-diacetate (DCF-DA) Expression of apoptosis signaling proteins Bcl-2, Bax and Caspase-3. Results: After incubated with 1.5μmol·L-1 rotenone for 24 h, the viability of PC12 cells was significantly decreased. Baicalin inhibited the cell injury induced by rotenone in a dose-dependent manner. Flow cytometry analysis showed that baicalin significantly reduced rotenone Induced apoptosis of PC12 cells. Fluorescence microscopy showed that baicalin could significantly reduce the ROS content of PC12 cells damaged by rotenone. Western blotting showed that baicalin up-regulated the expression of Bcl-2 and down-regulated the expression of Bax and decreased the activation of Caspase-3 in model cells. CONCLUSION: Baicalin has a protective effect on rotenone-induced PC12 injury. Its mechanism may be related to the decrease of ROS content of rotenone-induced PC12 cells and inhibition of mitochondrial apoptosis pathway induced by rotenone.