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目的探讨哈巴苷对β淀粉样蛋白(Aβ_(25-35))诱导的PC12细胞毒性的影响。方法 PC12细胞以哈巴苷(20、40、80μmol/L)预孵育3 h后以Aβ25-35(30μmol/L)损伤24 h;MTT法检测细胞增殖;检测细胞上清液中丙二醛(MDA)、谷胱甘肽(GSH)、超氧化物歧化酶(SOD)水平;采用Annexin-V-FITC双染法检测细胞凋亡率;检测活性氧(ROS)的生成;Western blotting法检测Akt、Bcl-2和Bax蛋白表达。结果与对照组比较,Aβ25-35使PC12细胞的存活率显著降低(P<0.01),细胞上清液中MDA水平显著升高(P<0.01)、GSH和SOD水平显著降低(P<0.01),细胞内ROS水平显著升高(P<0.01),细胞凋亡率显著增加(P<0.01),细胞内p-Akt、Bcl-2蛋白表达下调(P<0.01),Bax蛋白表达显著上调(P<0.01);哈巴苷预孵育PC12细胞3 h,使细胞上清液中的MDA水平降低,GSH和SOD水平显著升高(P<0.05、0.01);明显剂量依赖性抑制细胞凋亡;上调p-Akt和Bcl-2蛋白表达(P<0.05、0.01),并下调Bax蛋白表达(P<0.05、0.01)。结论哈巴苷改善Aβ25-35诱导的PC12细胞毒性,可能通过激活PI3K/Akt信号通路和上调细胞内SOD水平发挥作用。
Objective To investigate the effect of habituoside on PC12 cytotoxicity induced by β-amyloid protein (Aβ_ (25-35)). Methods PC12 cells were pre-incubated with habacin (20, 40, and 80μmol / L) for 3 h and then harvested for 24 h with Aβ25-35 (30 μmol / L). Cell proliferation was measured by MTT assay. Malondialdehyde ), Glutathione (GSH) and superoxide dismutase (SOD) were detected by flow cytometry. Apoptosis rate was detected by Annexin-V-FITC double staining method. ROS generation was detected by Western blotting. Akt, Bcl-2 and Bax protein expression. Results Compared with the control group, Aβ25-35 significantly decreased the survival rate of PC12 cells (P <0.01), the level of MDA in cell supernatant increased significantly (P <0.01) and the levels of GSH and SOD decreased significantly (P <0.01) (P <0.01). The apoptotic rate was significantly increased (P <0.01), the expressions of p-Akt and Bcl-2 were down-regulated and the protein expression of Bax was up-regulated P <0.01). The preincubation of PC12 cells with habacin for 3 h reduced the levels of MDA in the supernatant and the levels of GSH and SOD (P <0.05, 0.01), and significantly inhibited the apoptosis in a dose-dependent manner p-Akt and Bcl-2 protein (P <0.05, 0.01), and down-regulated Bax protein expression (P <0.05, 0.01). Conclusion Habasone can improve Aβ25-35-induced PC12 cytotoxicity possibly through activation of PI3K / Akt signaling pathway and up-regulation of intracellular SOD.