阿托伐他汀通过抑制ERK/MAPK信号通路改善H2O2诱导的血管内皮细胞损伤的机制研究

目的探究阿托伐他汀通过调节ERK/MAPK信号通路改善H₂O₂诱导的血管内皮细胞损伤的作用机制。方法体外培养人脐静脉内皮细胞(HUVECs),分为空白对照组、模型组(H₂O₂处理)、阿托伐他汀组(阿托伐他汀+H₂O₂)、U0126组(ERK/MAPK信号通路抑制剂U0126+H₂O₂)、阿托伐他汀+LM22B-10组(阿托伐他汀和ERK/MAPK信号通路激活剂LM22B-10+H₂O₂)。采用MTT法、AO/EB染色法分别检测各组细胞活性、凋亡率;用酶联免疫吸附法检测各组细胞乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)含量;荧光法检测活性氧簇(ROS)水平;Western blot法检测各组细胞中ERK1/2、p-ERK1/2、p38 MAPK、p-p38 MAPK蛋白表达。结果与空白对照组比较,模型组细胞增殖活性、SOD和GSH-Px含量降低,细胞凋亡率、LDH、MDA和ROS水平增加,p-ERK1/2/ERK1/2、p-p38 MAPK/p38 MAPK蛋白表达比值上调(P均<0.05)。与模型组比较,阿托伐他汀组细胞增殖活性、SOD和GSH-Px含量升高,细胞凋亡率、LDH、MDA和ROS水平降低,且p-ERK1/2/ERK1/2、p-p38 MAPK/p38 MAPK蛋白表达比值下调(P均<0.05)。与阿托伐他汀组比较,阿托伐他汀+LM22B-10组细胞增殖活性、SOD和GSH-Px含量降低,凋亡率、LDH、MDA和ROS水平升高,同时p-ERK1/2/ERK1/2、p-p38 MAPK/p38 MAPK蛋白表达比值上调(P均<0.05)。结论阿托伐他汀能够改善H₂O₂诱导的血管内皮细胞损伤,其机制与抑制ERK/MAPK信号通路进而降低ROS介导的血管内皮细胞凋亡和氧化应激损伤有关。

Mechanism study of atorvastatin ameliorates H2O2 induced vascular endothelial cell injury by inhibiting ERK/MAPK signaling pathway

Objective To explore the underlying mechanism by which atorvastatin ameliorates H₂O₂ induced vascular endothelial cell injury by regulating the ERK/MAPK signaling pathway.Methods Human umbilical vein endothelial cells(HUVECs)were cultured in vitro and divided into blank control group(conventional culture),model group(treated with H₂O₂),atorvastatin group(treated with atorvastatin+H₂O₂),U0126 group(treated with ERK/MAPK inhibitor U0126+H₂O₂),and atorvastatin+LM22B-10 group(treated with atorvastatin and ERK/MAPK activator LM22B-10+H₂O₂).Cell proliferation and apoptosis were detected by MTT and AO/EB staining methods.The levels of lactate dehydrogenase(LDH),malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione peroxidase(GSH PX)were detected by ELISA kit.The level of reactive oxygen species(ROS)was detected by fluorescence method.The expression of p-ERK1/2,ERK1/2,p38 MAPK,p-38 MAPK proteins were detected by Western blot.Results Compared with the blank control group,the cell proliferation activity,SOD and GSH-Px content were significantly decreased in the model group,meanwhile the apoptosis rate,MDA,LDH,ROS levels and p-ERK1/2/ERK1/2 ratio and p-p38 MAPK/p38 MAPK ratio were significantly increased(all P<0.05).Compared with the model group,the cell proliferation activity,SOD and GSH-Px content were significantly increased in the atorvastatin and U0126 groups,meanwhile the apoptosis rate,MDA,LDH,ROS levels and p-ERK1/2/ERK1/2 ratio or p-p38 MAPK/p38 MAPK ratio were significantly decreased(all P<0.05).However,compared with the atorvastatin Group,the cell proliferation activity,SOD and GSH-Px contents in the atorvastatin+LM22B-10 group were decreased,and the apoptosis rate,LDH,MDA and Ros levels were increased,at the same time,p-ERK1/2/ERK1/2 and p-p38 MAPK/p38 MAPK protein expression ratios were up-regulated(all P<0.05).Conclusion Atorvastatin can improve vascular endothelial cell damage,and its mechanism may be related to the inhibition of ERK/MAPK signaling pathway and HUVECs apoptosis and oxidative stress injury induced by ROS,thus maintaining the cell viability.