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研究水飞蓟素预处理对丙烯酰胺诱导肝细胞凋亡的抑制作用,并探讨其作用机制。以Hep G2细胞为模型细胞,设4个水飞蓟素预处理组(12,24,48,96μg/m L),预处理时间6 h,用8 mol/L丙烯酰胺染毒;同时设1个丙烯酰胺染毒组和1个细胞悬液对照组。分别采用MTT法和Annexin V-FITC双染法检测细胞活力和凋亡情况;采用细胞化学法检测细胞中Caspase-3的活性及ROS和GSH水平。结果表明,丙烯酰胺可显著增加细胞凋亡率,降低细胞存活率(P<0.01)。用12~96μg/m L质量浓度范围的水飞蓟素对细胞进行预处理后,细胞凋亡率明显降低,存活率明显提高;同时细胞内ROS水平和Caspase-3的活性显著降低,细胞内GSH含量明显提高(P<0.01),且在水飞蓟素质量浓度为96μg/m L时,达到最佳效果。以上说明水飞蓟素可有效抑制丙烯酰胺诱导的肝细胞凋亡,原因可能与其减缓细胞内氧化压力和抑制肝细胞凋亡途径中关键酶Caspase-3活性有密切关系。
To study the inhibitory effect of silymarin pretreatment on the apoptosis induced by acrylamide in hepatocytes and to explore its mechanism. Hep G2 cells were used as model cells. Four silymarin pretreatment groups (12, 24, 48 and 96 μg / mL) were pretreated with 8 mol / L acrylamide for 6 h. At the same time, one acrylamide Exposure groups and 1 cell suspension control group. Cell viability and apoptosis were detected by MTT assay and Annexin V-FITC double staining respectively. Caspase-3 activity and ROS and GSH levels were detected by cytochemistry. The results showed that acrylamide significantly increased the apoptosis rate and decreased the cell survival rate (P <0.01). Pretreatment of cells with silymarin in the concentration range of 12 ~ 96μg / m L significantly decreased the apoptosis rate and significantly increased the survival rate. At the same time, the levels of intracellular ROS and Caspase-3 activity were significantly decreased, and the intracellular GSH content was significantly increased (P <0.01), and the best effect was achieved when the silymarin concentration was 96μg / m L. These results indicate that silymarin can effectively inhibit the apoptosis of hepatocytes induced by acrylamide, which may be related to its role in alleviating the intracellular oxidative stress and inhibiting the activity of Caspase-3, a key enzyme in the hepatocyte apoptosis pathway.