益肾方对大鼠肾小管上皮细胞氧化应激的保护作用及其机制研究

Research on the Protective Effect and the Mechanism of Yishen Decoction on Oxidative Stress in Renal Tubular Epithelial Cells

目的:探究益肾方对NRK-52E细胞氧化损伤的保护作用。方法:建立H_2O_2诱导NRK-52E氧化损伤模型,分为正常对照组,H_2O_2模型组,益肾方组。流式细胞术测定细胞凋亡率,运用Western-blot技术检测Caspase-3和BCL-2的蛋白表达。结果:与正常对照组比较,H_2O_2造模组的细胞凋亡率明显增高(P <0. 01);与H_2O_2模型组比较,益肾方组在益肾方浓度100μg/ml、200μg/ml、400μg/ml时,细胞的凋亡率出现明显的下降(P <0. 05)。与H_2O_2模型组相比,益肾方组在益肾方浓度100μg/ml、200μg/ml时,Caspase-3的蛋白表达明显降低,BCL-2的蛋白表达增加(P <0. 05)。结论:益肾方对NRK-52E细胞氧化损伤的保护机制,可能与其降低细胞凋亡,同时下调Caspase-3和上调BCL-2的蛋白表达有关。

Objective: To explore the protection mechanisms of Yishen Decoction on oxidative damaged renal tubular epithelial cells of rat( NRK-52 E). Methods: To establish the model of oxidative damage in NRK-52 E cells,Experimental groups were divided into normal control group,H2 O2 model group and Yishen Decoction group. The apoptosis rate was determined by Flow cytometry. The protein expression of Caspase-3 and BCL-2 was detected by Western-blot. Results: Compared with normal controls,the apoptosis rate of the H2 O2 model group was significantly higher( P < 0. 01). Compared with the H2 O2 model group,the apoptosis rate of the cells was significantly decreased( P < 0. 05) when the drug concentration of Yishen Decoction was 100 μg/ml,200 μg/ml and 400 μg/ml. Compared with the H2 O2 model group,the protein expression of Caspase-3 in Yishen Decoction group decreased significantly,when concentration is 100 μg/ml,200 μg/ml,the protein expression of BCL-2 increased,there were statistical differences( P < 0. 05). Conclusion: The protective mechanism of Yishen Decoction for the oxidative damage of NRK-52 E may be associated with lower cell apoptosis by down-regulation the protein Expression of Caspase-3 and up-regulation the protein Expression of BCL-2.