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目的:观察珠子参总皂苷对H2O2致心肌细胞氧化应激性损伤保护作用及MCP-1和相关炎症因子表达的影响。方法:Wistar乳鼠心肌细胞原代培养,用H2O2建立氧化应激损伤模型,然后用珠子参总皂苷(100,200μg/ml)孵育24 h后,MTT法检测珠子参总皂苷对心肌细胞活力的影响,Hoechst33258染色检测细胞内ROS含量的影响,比色法测定培养液中LDH、CK含量,同时检测培养液中SOD、CAT、GSH-Px的活性及MDA含量,ELISA检测培养液中MCP-1、TNF-α、TGF-β1含量,Western blot检测细胞内MCP-1、NF-κBp65蛋白表达。结果:珠子参总皂苷(100,200μg/ml)处理能显著增加存活率,改善细胞形态,降低细胞内ROS含量;减少LDH、CK释放量,增强SOD、CAT和GSH-Px活性,降低MDA含量;降低培养液中MCP-1、TNF-α、TGF-β1含量及细胞内MCP-1、NF-κBp65蛋白表达水平,其中以珠子参总皂苷200μg/ml剂量组效果更为显著。结论:珠子参总皂苷对H2O2致心肌细胞氧化损伤具有保护作用,其保护作用主要与抑制心肌细胞内ROS的产生与聚集,增强抗氧化酶的活性,减少心肌酶的释放及抑制MCP-1、TNF-α、TGF-β1和NF-κB p65的表达有关。
OBJECTIVE: To observe the protective effect of total saponins from Bentley Roots induced by H2O2 on the oxidative stress induced injury of cardiomyocytes and the expression of MCP-1 and related inflammatory factors. Methods: Wistar neonatal rat cardiomyocytes were cultured in primary culture. H2O2 was used to establish the model of oxidative stress injury. Then the cells were incubated with total ginsenosides (100, 200μg / ml) for 24 h. The effect of total ginsenosides on the viability of cardiomyocytes was detected by MTT assay , Hoechst33258 staining. The content of LDH and CK in the culture medium was measured by colorimetric assay. The activities of SOD, CAT, GSH-Px and the contents of MDA in the culture medium were detected by ELISA. The levels of MCP-1, TNF-α and TGF-β1 were detected by Western blot. The expression of MCP-1 and NF-κB p65 protein were detected by Western blot. Results: Total ginsenosides (100, 200μg / ml) could significantly increase the survival rate, improve the cell morphology, decrease the content of intracellular ROS, decrease the release of LDH and CK, increase the activity of SOD, CAT and GSH-Px and decrease the content of MDA. The content of MCP-1, TNF-α, TGF-β1 in the culture medium and the expression of MCP-1 and NF-κBp65 in the culture medium were decreased. The effect of the total ginsenoside 200 μg / ml group was more significant. CONCLUSION: The total saponins of Beizhuanbao have the protective effect on the oxidative damage of cardiomyocytes induced by H2O2. Its protective effect is mainly related to the inhibition of the production and accumulation of ROS in myocardial cells, enhancing the activity of antioxidant enzymes, reducing the release of myocardial enzymes and inhibiting the expression of MCP-1, TNF-α, TGF-β1 and NF-κB p65 expression.