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目的:探讨金银花水提物对过氧化氢(H_2O_2)诱导的大鼠肾上腺嗜铬细胞瘤PC12细胞氧化损伤的保护作用。方法:采用H_2O_2损伤PC12细胞建立氧化损伤模型,以不同浓度金银花水提物(5、25和50μg/ml)进行研究,采用MTT法检测细胞活力,DCFH-DA检测ROS的水平,TUNEL法检测细胞凋亡,免疫印迹实验检测细胞中Sirt3和凋亡相关蛋白蛋白表达。结果:不同浓度金银花水提物(5、25和50μg/ml)能够抑制H_2O_2诱导的细胞死亡并呈剂量依赖性。H_2O_2组细胞活力和凋亡率较对照组显著增加,给予金银花水提物作用48 h后,明显降低细胞内活性氧(ROS)和丙二醛(MDA)含量,增加超氧化物歧化酶(SOD)活力,抑制H_2O_2诱导的细胞凋亡并上调Sirt3、IDH2、Bcl-2蛋白表达,下调Bax、caspase3蛋白表达。结论:金银花水提物能够抑制H_2O_2诱导的PC12细胞的氧化损伤,其保护作用可能与抑制氧化应激引起的细胞凋亡并调控ROS/Sirt3途径有关。
Objective: To investigate the protective effect of aqueous extract of honeysuckle (Honeysuckle) on oxidative damage of PC12 cells induced by hydrogen peroxide (H_2O_2) in rat adrenal pheochromocytoma. Methods: Oxidative injury was induced by H_2O_2 in PC12 cells. The cell viability was detected by MTT assay at different concentrations (5, 25 and 50μg / ml). ROS was detected by DCFH-DA. Apoptosis and Western blotting were used to detect the expression of Sirt3 and apoptosis related proteins in the cells. Results: Different concentrations of aqueous extract of honeysuckle (5, 25 and 50μg / ml) could inhibit H 2 O 2 -induced cell death in a dose-dependent manner. Compared with the control group, the viability and apoptosis rate of H 2 O 2 group were significantly increased. After being treated with the aqueous extract of Honeysuckle for 48 h, the content of reactive oxygen species (ROS) and malondialdehyde (MDA) in H 2 O 2 group was significantly decreased and the activity of superoxide dismutase (SOD) ), Inhibited the apoptosis induced by H_2O_2, up-regulated the expression of Sirt3, IDH2 and Bcl-2, and down-regulated the expression of Bax and caspase3. Conclusion: Honeysuckle extract can inhibit H 2 O 2 -induced oxidative damage of PC12 cells, and its protective effect may be related to the inhibition of apoptosis induced by oxidative stress and the regulation of ROS / Sirt3 pathway.