辛伐他汀抑制缺氧条件下人视网膜色素上皮细胞HIF-1α和VEGF的表达水平

Simvastatin inhibits HIF-1αand VEGF expression in RPE cells under hypoxia conditions

目的:探讨缺氧条件下辛伐他汀(Sim)对体外培养的人视网膜色素上皮(RPE-19)细胞的影响及其可能机制。方法:将体外培养的人RPE-19细胞随机分为三组:对照组、缺氧组(培养基中CoCl_(2)的最终浓度为125μmol/L)和Sim处理组(在含有125μmol/L CoCl_(2)的RPE-19细胞培养基中加入3μmol/L Sim)。24h后,观察RPE-19细胞的形态,用MTT法检测细胞增殖,采用酶联免疫吸附试验(ELISA)和免疫印迹法检测缺氧诱导因子1-α(HIF-1α)和血管内皮生长因子(VEGF)的分泌水平和蛋白表达,采用免疫印迹法检测自噬蛋白的表达水平,TUNEL法检测细胞凋亡。结果:缺氧条件下RPE-19细胞的形态发生了明显变化。缺氧组RPE-19细胞中HIF-1α和VEGF的蛋白表达明显增加,Sim治疗后显著降低。Beclin1和LC3B蛋白在CoCl_(2)+Sim组中的表达水平有所下降,且表达水平显著低于对照组和CoCl_(2)组。缺氧条件下,Sim抑制了RPE-19细胞增殖,促进了细胞的凋亡。结论:Sim可抑制缺氧条件下RPE-19细胞HIF-1α和VEGF蛋白的表达,抑制细胞增殖,促进凋亡,Sim促进RPE-19细胞凋亡的机制可能与其抑制自噬有关。

AIM:To investigate the effects of simvastatin(Sim)on human retinal pigment epithelial cells(RPE-19)and the possible mechanisms in vitro under hypoxia.METHODS:RPE-19 cells were divided into three group:control group,hypoxia group(the final concentration of CoCl_(2) in the medium was 125μmol/L),and Sim treatment group(3μmol/L Sim was added in the RPE-19 cells’medium which contain 125μmol/L CoCl_(2)).After 24h,the morphology of RPE-19 cells were observed,the proliferation of cells were calculated by MTT,the secretion levels and protein expression of hypoxia-inducible factor 1-Alpha(HIF-1α)and vascular endothelial growth factor(VEGF)were detected by enzyme-linked immunosorbent assay(ELISA)and Western blotting.The expression level of autophagy protein was detected by Western blot and apoptosis was detected by TUNEL.RESULTS:The morphology and activity of RPE-19 cells showed an apparent change under hypoxia.The expression of HIF-1αand VEGF protein were increased obviously in the hypoxia group and then significantly decreased after Sim treatment.Beclin1,and LC3B proteins were decreased in the CoCl_(2)+Sim group,and the expression levels were lower than the control and CoCl_(2)group.Under hypoxia,Sim inhibited RPE-19 cells’proliferation and promoted the apoptosis.CONCLUSION:Sim inhibits RPE-19 cells’proliferation,decreases HIF-1αand VEGF protein,and promotes apoptosis under hypoxia.Our results suggested that the mechanism by which Sim promoted apoptosis in RPE-19 cells may be related to its inhibition of autophagy.