二甲双胍通过激活AMPK降低同型半胱氨酸对内皮细胞的损伤  

Metformin Attenuates Endothelial Cell Injury Caused by Homocysteine Through AMPK Pathway

作　　者：付勇南[1] 胡金兴 杨媛[2] 王梦洪[1] 郑泽琪[1] 彭景添[1] 

机构地区：[1]南昌大学第一附属医院心内科,江西省南昌市330006 [2]南昌大学第一附属医院药学部,江西省南昌市330006

出　　处：《中国动脉硬化杂志》2016年第11期1081-1085,共5页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金项目(81260049)

摘　　要：目的探讨二甲双胍能否降低同型半胱氨酸(Hcy)对血管内皮细胞的损伤及其可能的机制。方法培养人血管内皮细胞株(EC304人血管内皮细胞),分为对照组、Hcy组、二甲双胍组和Hcy+二甲双胍组,采用CCK-8检测细胞活力,采用乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、丙二醛(MDA)试剂盒测定内皮细胞LDH、SOD活性及细胞内MDA含量,采用RT-PCR检测SOD1、过氧化氢酶(CAT)和NADPH氧化酶2(NOX2)mRNA的表达,采用Western blot检测AMPKα和p-AMPKα蛋白的表达。结果与对照组相比,Hcy组内皮细胞活力明显下降,LDH活性增加,细胞内MDA含量升高,SOD活性下降,SOD1和CAT mRNA表达水平显著下降,而NOX2 mRNA表达水平则明显升高,二甲双胍能够抑制Hcy引起的细胞活力下降及LDH、MDA增加;二甲双胍增加SOD活性,增加SOD1和CAT mRNA表达水平,减少NOX2 mRNA表达水平。AMPK抑制剂Compoud C则可以逆转二甲双胍对Hcy引起的内皮细胞损伤的保护作用。结论二甲双胍激活AMPK可能通过调控细胞内氧化应激抑制Hcy对内皮细胞的损伤。Aim To investigate the protective effects of metformin on vascular endothelial cells injured by homocysteine（Hcy） and its mechanism. Methods Human vascular endothelial cell line（ECV304 cells） cultivated in vitro were divided into four groups： control group,Hcy group,metformin group,Hcy and metformin co-incubation group,the cell proliferation ability was determined by CCK-8 assay. The lactate dehydrogenase（LDH）,the content of malondialdehyde（MDA） and superoxidase dismutase（SOD） activity in ECV304 cells cellular supernatant were detected. RTPCR was performed to detect mRNA expression of SOD1. p-AMPKα and t-AMPKα protein expression was detected by Western blot. Results Homocysteine significantly inhibited endothelial cell viability and increased the activities of LDH. Compared with the control group,the content of MDA increased and the activity of SOD decreased in Hcy group.Metformin significantly improved endothelial cell viability and SOD activity,suppressed homocysteine induced increases in LDH and MDA. Homocysteine decreased the expression of SOD1 and catalase（CAT） mRNA,increased NADPH oxidase 2（NOX2） mRNA. Metformin increased the expression of SOD1 and CAT mRNA,decreased NOX2 mRNA compared with Hcy group. Compoud C（an AMPK inhibitor） could reverse the protection of metformin. Conclusion Metformin could protect the ECV304 cells from injury by homocysteine via AMPK-ROS signalling pathway.

关 键 词：二甲双胍 同型半胱氨酸 腺苷酸活化蛋白激酶 内皮细胞损伤 氧化应激 

分 类 号：R363[医药卫生—病理学]