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目的:探讨白藜芦醇对氧糖剥夺/再灌注(OGD/R)损伤的PC12细胞的保护作用及其机制。方法:体外培养PC12细胞,分为对照组,白藜芦醇组,OGD/R组及OGD/R+白藜芦醇组。以改良的噻唑蓝法测定细胞活性,采用Annexin V-FITC/PI双染法检测细胞的凋亡率,用双氢罗丹明(DHR)检测细胞内活性氧簇(ROS)的水平,采用蛋白印迹法(western blot)分析SIRT1的蛋白表达情况。结果:与对照组相比,经过OGD/R损伤后,细胞活力显著降低。而在OGD/R的同时给予10μmol/L的白藜芦醇处理,可以明显提高细胞活力。流式细胞仪检测发现,10μmol/L的白藜芦醇可以显著地减少OGD/R引起的细胞凋亡,抑制细胞内的ROS产生。western blot的结果提示,与对照组比较,白藜芦醇可提高SIRT1的蛋白表达水平。结论:白藜芦醇可以通过抑制ROS的产生和上调SIRT1的表达等机制而发挥其对抗氧糖剥夺/再灌注损伤的神经保护性作用。
Objective: To investigate the protective effect of resveratrol on PC12 cells injured by oxygen deprivation / reperfusion (OGD / R) and its mechanism. Methods: PC12 cells were cultured in vitro and divided into control group, resveratrol group, OGD / R group and OGD / R + resveratrol group. Cell viability was determined by modified thiazolyl blue staining. Cell apoptosis was detected by Annexin V-FITC / PI double staining. Intracellular reactive oxygen species (ROS) levels were detected by DHR. Western blot Western blot analysis of SIRT1 protein expression. Results: After OGD / R injury, cell viability was significantly reduced compared with the control group. In OGD / R given 10μmol / L of resveratrol, can significantly improve cell viability. Flow cytometry showed that 10μmol / L resveratrol could significantly reduce the apoptosis induced by OGD / R and inhibit the intracellular ROS production. The results of western blot suggested that resveratrol could increase the protein expression of SIRT1 compared with the control group. CONCLUSION: Resveratrol can exert its neuroprotective effect against OGD / reperfusion injury by inhibiting the production of ROS and up-regulating the expression of SIRT1.