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

中国临床药理学与治疗学 ›› 2015, Vol. 20 ›› Issue (1): 14-18.

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

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

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,汕头 515041,广东

收稿日期:2014-09-21 修回日期:2014-10-23 发布日期:2020-07-20
通讯作者: 石刚刚,通信作者,男,博士,教授,博士生导师,研究方向:开发抗心肌缺血药物,生物医药与先进材料研究。Tel:0754-88900301 E-mail:ggshi@stu.edu.cn
作者简介:周燕琼,女,博士,副主任药师,研究方向:开发抗心肌缺血药物,医院药学,临床药学。Tel:0754-88915672 E-mail:styqzhou@163.com
基金资助:
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);中央财政支持地方高校发展专项资金;国家自然科学基金资助项目(81173048,81072633,30901810);汕头市科技计划项目(汕府科[2014)62号]

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

ZHOU Yan-qiong^1,2, ZHANG Yan-mei¹, GAO Fen-fei¹, HUANG Zhan-qin¹, CHEN Yi-cun¹, ZHENG Yan-shan¹, SHI Gang-gang¹

¹Department of Pharmacology,
²Second Affiliated Hospital, Shantou University Medical College, Shantou 515041, Guangdong, China

Received:2014-09-21 Revised:2014-10-23 Published:2020-07-20

摘要/Abstract

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。

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

Abstract: AIM: To investigate the effects of antisense oligodeoxynucleotides (AS-ODN) on Egr-1 protein expression in cultured endothelial cells after anoxia/reoxygenation (A/R). METHODS: The cultured cardiac microvascular endothelial cells (CMECs) A/R model were established. The cells were randomly divided into one of six groups: Con, A/R, Lip, AS, S, and Sc. Levels of lactate dehydrogenase (LDH), superoxide 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 analyses. Cell morphology and cell viability were observed to assess the degree of injury. RESULTS: Treatment with Egr-1 AS-ODN significantly reduced 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-α 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.

Key words: antisense oligodeoxynucleotides, cardiac microvascular endothelial cells, anoxia/ reoxygenation injury, Egr-1

中图分类号:

R965.2

周燕琼, 张艳美, 高分飞, 黄展勤, 陈一村, 郑燕珊, 石刚刚. 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[J]. 中国临床药理学与治疗学, 2015, 20(1): 14-18.

ZHOU Yan-qiong, ZHANG Yan-mei, GAO Fen-fei, HUANG Zhan-qin, CHEN Yi-cun, ZHENG Yan-shan, SHI Gang-gang. The protective effects of Egr-1 antisense oligodeoxynucleotides on endothelial injury induced by anoxia-reoxygenation[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2015, 20(1): 14-18.

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