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目的探讨补骨脂素对H_2O_2诱导PC-12细胞氧化损伤的保护作用,并考察其作用机制。方法 PC-12细胞分为对照组、模型组、Tempol组和补骨脂素组。对照组中未加入补骨脂素和H_2O_2;模型组中加入400μmol/L H_2O_2;Tempol组中加入100μmol/L Tempol及400μmol/L H_2O_2;补骨脂素组中加入400μmol/L H_2O_2及1、5、10、20μmol/L补骨脂素。CCK-8法测定PC-12细胞的存活率,考察对PC-12细胞形态的影响,PI/Annexin-V、Hoechst染色检测细胞凋亡率,蛋白质印迹法检测凋亡蛋白表达。结果 5、10、20μmol/L补骨脂素均可以显著提高PC-12细胞内的吸光度值(P<0.01),细胞存活率提高较明显,与模型组比较差异具有统计学意义(P<0.05)。补骨脂素1、5、10、20μmol/L组凋亡率明显低于模型组(P<0.05),并且随着补骨脂素浓度的增加,PC-12细胞的凋亡率逐渐下降。随着补骨脂素作用时间的增加,各组PC-12细胞凋亡率的增加幅度相近,但明显低于模型组(P<0.05)。随着补骨脂素浓度的增加,对磷酸化Bad、Bcl-XL水平的上调作用越明显。结论补骨脂素对H_2O_2诱导PC-12细胞损伤具有保护作用,其作用机制是通过上调磷酸化Bad和Bcl-XL表达来抑制细胞凋亡。
Objective To investigate the protective effect of psoralen on the oxidative damage induced by H 2 O 2 in PC-12 cells and to investigate its mechanism. Methods PC-12 cells were divided into control group, model group, Tempol group and psoralen group. Psoralen and H 2 O 2 were not added in the control group; 400 μmol / L H 2 O 2 was added into the model group; 100 μmol / L Tempol and 400 μmol / L H 2 O 2 were added into the Tempol group; 400 μmol / L H 2 O 2 and 1,5 , 10,20 μmol / L psoralen. The survival rate of PC-12 cells was determined by CCK-8 assay. The morphological changes of PC-12 cells were examined. PI / Annexin-V and Hoechst staining were used to detect the apoptosis rate. Western blotting was used to detect the expression of apoptotic protein. Results Psoralen at 5, 10 and 20μmol / L significantly increased the absorbance of PC-12 cells (P <0.01), and the cell viability increased more significantly than that of model group (P <0.05) ). The apoptosis rates of psoralen in 1,5,10,20 μmol / L group were significantly lower than those in model group (P <0.05), and the apoptosis rate of PC-12 cells decreased with the increase of psoralen concentration. With the increase of psoralen time, the increase rate of PC-12 cell apoptosis in each group was similar, but significantly lower than that in model group (P <0.05). With the increase of psoralen concentration, the up-regulation effect on phosphorylation of Bad and Bcl-XL is more obvious. Conclusion Psoralen can protect PC-12 cells from injury induced by H 2 O 2, and its mechanism is that apoptosis is inhibited by up-regulating the expressions of Bad and Bcl-XL.