青蒿琥酯对体外培养的视网膜神经节细胞氧化应激损伤的保护机制研究

目的:研究青蒿琥酯对体外培养的视网膜神经节细胞氧化应激损伤的保护作用及其可能的作用机制。方法:体外培养视网膜神经节RGC-5细胞，将细胞分为空白对照组、H2O2组、H2O2+低剂量青蒿琥酯组、H2O2+中剂量青蒿琥酯组、H2O2+高剂量青蒿琥酯组，CCK8法检测各组细胞的相对存活率，流式细胞术检测细胞凋亡率，试剂盒检测各组细胞匀浆中氧化应激产物MDA、NO、SOD1水平，Western blot检测各组细胞中HO-1、SOD1、PI3K、Akt及p-Akt蛋白表达水平。结果:与H2O2组比较，不同剂量青蒿琥酯可以促使RGC-5细胞增殖，抑制其凋亡，差异比较有统计学意义（P<0.05）；各药物组中MDA、NO显著低于H2O2组，而SOD1活性显著高于H2O2组；各药物组HO-1、SOD1、PI3K及p-Akt蛋白表达水平均显著高于H2O2组，差异均有统计学意义（P<0.05）；并且PI3K抑制剂能降低青蒿琥酯对RGC-5细胞的保护作用。结论:青蒿琥酯能明显降低H2O2诱导的视网膜神经节细胞氧化应激损伤，其作用机制可能与激活PI3K/Akt信号通路有关。

Effects of artesunate on oxidative stress in cultured retinal ganglion cells and its protective mechanism

AIM: To study the effects of artesunate on oxidative stress in cultured retinal ganglion cells (RGC-5) and to investigate its possible mechanism. METHODS: RGC-5 cells were cultured in vitro and were divided into five groups, including control group, H2O2 group, low-, middle-, high-dose artesunate groups. CCK8 assay was used to measure the relative survival rate of RGC-5 cells. Cell apoptotic rate was determined by flow cytometry. Related kits were used to determine the levels of MDA, NO and SOD1. Western bot method was used to analyze the expression levels of HO-1, SOD1, PI3K, p-Akt and Akt. RESULTS:Compared with H2O2 group, the relative survival rates were significantly increased and cell apoptotic rates were greatly decreased in varying concentrations of artesunate groups (P<0.05). The contents of MDA, NO were greatly decreased, while the activity of SOD1 was strongly increased in artesunate groups compared with H2O2 group (P<0.05). The expression levels of HO-1, SOD1, PI3K and p-Akt were greatly increased in artesunate groups compared with H2O2 group (P<0.05). Moreover, PI3K inhibitor decreased the protective effect of artesunate on the injury of RGC-5 cells. CONCLUSION: Artesunate significantly inhibit the oxidative stress injury of RGC-5 cells and its underlying mechanism maybe related with inducing the activation of PI3K/Akt signaling pathway.