黄芪甲苷对过氧化氢诱导动脉平滑肌细胞凋亡的抑制作用和机制探讨-药理学专业毕业论文.docxVIP

PAGE PAGE 10 组和保护细胞的羟乙基葛根素组中的细胞形态相似, 说明 AST 可以很好地保护 细胞结构, 维持细胞的正常形态, 以上各组细胞 DNA 片段化分析均无凋亡“梯 形条带”出现。 最后 Western blot 结果显示 AST 上调 Bcl-2、下调 Bax、p-Jak2 和 Caspase-9 蛋白表达, 为了阻断 Jak2 磷酸化为 p-Jak2, 我们先用 100μmol/L AG490 预处理细胞 12h, 然后照以上同样处理, 结果各实验组中 p-Jak2 蛋白表达 大幅下降, 接近正常组水平, 提示 Jak2 可能是 AST 发挥抗氧化药效的作用靶点。 结论：AST 有抑制 H202 诱导动脉平滑肌细胞凋亡的作用, AST 的保护机制与上 调 Bcl-2, 下调 Bax、p-Jak2 和 Caspase-9 蛋白表达有关。 关键词：黄芪甲苷 过氧化氢 动脉平滑肌细胞 凋亡 Bcl-2 Bax p-Jak2 Caspase-9 AG490 . Depressant effects of Astragaloside on aortic smooth muscle cells apoptosis induced by hydrogen dioxide and the mechanism involved ABSTRACT Objective: Based on the observation of the ASMCs apoptosis induced by hydrogen dioxide, this experiment was aimed to investigate the depressant effects of AST on the aortic smooth muscle cells (ASMCs) treated with hydrogen dioxide and the pharmacological mechanism involved. Methods: The apoptotic model of ASMCs, induced by H202 at suitable concentration and treatment time, was constructed by determining cell survival rate with 3-(4,5-Dimethylthiazol-2yl)-2,5-Diphenyl Tetrazolium Bromide(MTT). According to respective requirement of the distinct group, the ASMCs, pretreated with 10、20、40、80、160μg/ml AST for 1 h , were dealed with 0.5mmol/L H202 for 6h, then, the effect of AST and H202 on the ASMCs was detected by several approaches, including DNA fragmentation analysis for the DNA degradation, acridine orange and ethidium bromide (AO/EB) staining for the change of cell shape, MTT and flow cytometry analysis with Propidum Iodide(PI) for the certain cell survival rate, eventually, Western blot for the diversify of the protein expression of Bcl-2、Bax、 p-Jak2 and Caspase-9. Pretreated with AG490, the special blocker of phosphorylation of Jak2, the change of expression of protein p-Jak2 was determined. Results: In the apoptotic mode of ASMCs 0.5mmol/L H202 induced for six hours, the cell survival rate reached 51.43±3.98%, the apoptotic cells increasing largely by AO/EB staining,