PI3K/Akt通路在芍药苷抗Aβ_(25-35)诱导PC12细胞凋亡中的作用

Role of PI3K / Akt pathway in effect of paeoniflorin against Aβ_(25-35)-induced PC12 cell injury

目的:研究PI3K/Akt通路在芍药苷对PC12细胞神经保护中的作用。方法:芍药苷组(5,10,20μmol·L-1)预处理30 min,然后加入Aβ25-35(20μmol·L-1)共同作用24 h;抑制剂LY294002(10μmol·L-1)于芍药苷(10μmol·L-1)作用前预处理30 min。用MTT比色法检测细胞存活率,Annexin-V/PI双染检测细胞凋亡率,以及Western blot法检测p-Akt,Bax,Bcl-2,cleaved caspase-3蛋白表达。结果:芍药苷可抑制Aβ25-35所诱导的PC12细胞毒性和凋亡,其保护作用可能是通过上调Akt磷酸化水平,增加Bcl-2蛋白表达、降低Bax蛋白表达、抑制caspase-3等的激活而实现的;抑制剂LY294002可减弱上述芍药苷的保护作用。结论:芍药苷可通过激活PI3K/Akt信号通路对Aβ25-35诱导PC12细胞损伤具有保护作用。

Objective： To study the role of PI3K/Akt pathway in the neuroprotective effect of paeoniflorin on PC12 cells. Method： The paeoniflorin group （5, 10, 20 μmol·L^-1） was pretreated for 30 min, and then added with Aβ25-35 （20 μmol·L^-1 for interaction for 24 h. Inhibitor LY294002 （10 μmol·L^-1） was pretreated for 30 min before the action of paeoniflorin （10 μmol·L^-1）. The MTT colorimetric method was used to detect the cell viability. The apoptosis rate was tested by the FITC-Annexin V/PI staining. The protein expression of p-AKT, Bax, Bcl-2 and cleaved caspase-3 protein were detected by Western blot analysis. Result： Paeoniflorin could significantly inhibit the Aβ25-35-induced PC12 cell toxicity and apoptosis. Its protection effect may be achieved by up-regulating AKT phosphorylation level, increasing Bcl-2 protein expression, reducing Bax protein expression, inhibiting the activation of caspase-3. Inhibitor LY294002 could weaken the above protective effects of paeoniflorin. Conclusion： Paeoniflorin could activate PI3K/Akt signaling pathway to protect the PC12 cell injury induced by Aβ25-35.