反义寡核苷酸抑制早期生长反应基因-1表达对内皮细胞缺氧复氧损伤的影响  

The protective effects of Egr-1antisense oligodeoxynucleotides on endothelial injury induced by anoxia-reoxygenation

作　　者：周燕琼[1,2] 张艳美[1] 高分飞[1] 黄展勤[1] 陈一村[1] 郑燕珊[1] 石刚刚[1] 

机构地区：[1]汕头大学医学院药理教研室 [2]汕头大学医学院第二附属医院

出　　处：《中国临床药理学与治疗学》2015年第1期14-18,共5页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金委员会-广东省人民政府自然科学联合基金资助项目(U0932005);国家自然科学基金资助项目(81173048;81072633;30901810);中央财政支持地方高校发展专项资金;汕头市科技计划项目(汕府科[2014)62号]

摘　　要：目的：研究反义寡核苷酸（antisense oligodeoxynucleotides,AS-ODN）抑制早期生长反应基因1（early growth response gene-1,Egr-1）表达对内皮细胞缺氧复氧（anoxia/reoxygenation,A/R）损伤的影响。方法：采用新生大鼠进行心脏微血管内皮细胞（cardiac microvascular endothelial cells,CMECs）原代培养,取3~4代的CMECs建立缺氧复氧模型。细胞随机分为6组：对照（Con）组、缺氧复氧组（A/R）、溶剂组（LIP）、反义寡核苷酸转染组（AS）、正义寡核苷酸转染组（S）和错配寡核苷酸转染组（Sc）。通过测定细胞培养上清液中乳酸脱氢酶（LDH）及内皮细胞中超氧化物歧化酶（SOD）、丙二醛（MDA）含量,观察内皮细胞损伤程度;应用ELISA法测定细胞培养上清液肿瘤坏死因子-α（TNF-α）的含量,观察内皮细胞炎症反应水平;Western-blot法检测培养内皮细胞中Egr-1的蛋白表达水平;显微镜下观察细胞形态学改变并计算存活率。结果：A/R造成内皮细胞内MDA升高,SOD下降,上清液中LDH、TNF-α含量升高,A/R刺激下细胞Egr-1的蛋白表达水平明显升高;A/R刺激前给予AS-ODN可抑制Egr-1蛋白的表达,减轻内皮细胞的损伤及炎症反应程度,提高细胞存活率。结论：AS-ODN抑制培养内皮细胞Egr-1的表达,并降低A/R损伤,提示Egr-1与缺氧复氧所致的心脏微血管内皮细胞损伤密切相关。AIM： To investigate the effects of antisense oligodeoxynucleotides （AS-ODN） on Egr-1 protein expression in cultured endothelial cells after anoxia/reoxygenation （A/R）. METH- ODS： The cultured cardiac microvaseular endo- thelial cells （CMECs） A/R model were estab- lished. The cells were randomly divided into one of six groups. Con, A/R, Lip, AS, S, and Sc. Levels of lactate dehydrogenase （LDH）, super- oxide dismutase （ SOD ）, malondialdehyde （MDA）, and tumor necrosis factor-α （TNF-α） were measured to assess the degree of injury and inflammation of endothelial cells. Egr-1 protein expression was examined by Western-blot analy- ses. Cell morphology and cell viability were ob- served to assess the degree of injury. RESULTS：Treatment with Egr-1 AS-ODN significantly re- duced Egr-1 protein expression and attenuated injury and inflammation of endothelial cells caused by A/R evidenced by the the decrease in leakage of LDH, the increase in SOD activity, the decrease in MDA generation, and release of TNF-~ from cultured CMECs. CONCLUSION： AS-ODN can protect cultured CMECs from A/R injury, by inhibiting the overexpression of Egr- 1. Egr-1 was related to the A/R injury of CMECs.

关 键 词：反义寡核苷酸 心脏微血管内皮细胞 缺 氧复氧损伤 EGR-1 

分 类 号：R965.2[医药卫生—药理学]