EPO预处理对培养心肌细胞缺氧/复氧损伤TNF-α表达的影响及机制被引量：1

Study of EPO pretreatment's effect on expression of TNF-α gene in cultured cardiac myocytes with hypoxia/reoxygenation injury and the possible mechanism

原文传递

导出

摘要目的观察促红细胞生成素(erythropoietin,EPO)预处理对培养心肌细胞缺氧复氧前后TNFα表达的影响,并进一步研究其可能的NFκB信号机制。方法采用培养乳鼠心肌细胞建立缺氧复氧损伤模型,分为对照组、EPO组[缺氧复氧前24h,培养液中加入终浓度10Uml的重组人促红素(recombinanthumanerythropoietin,RHuEPO)]、EPO+吡咯烷二硫氨基甲酸盐(PDTC)组(缺氧复氧前24h,加入终浓度10Uml的RHuEPO和5μgml的PDTC)和PDTC组(缺氧复氧前24h,加入终浓度5μgml的PDTC)。分别于缺氧复氧损伤前后,以RTPCR和westernblot检测各组心肌细胞TNFα基因表达变化,同时以EMSA检测各组心肌细胞NFκB活性变化。结果缺氧复氧损伤前,各组心肌细胞TNFα基因表达水平差异无统计学意义,均较弱。损伤后各组心肌细胞TNFα基因表达水平较损伤前对照组显著升高(P<0.01),而EPO组TNFα基因表达水平低于其他各组(P<0.01)。缺氧复氧损伤前EPO组NFκB活性高于其他各组(P<0.01),损伤后各组NFκB活性显著高于损伤前对照组(P<0.01),EPO组NFκB活性低于其他各组(P<0.01)。结论EPO预处理抑制缺氧复氧后心肌细胞TNFα基因表达升高,可能与缺氧复氧后NFκB活性升高被抑制有关。EPO预处理可能通过NFκB活化的负反馈机制抑制缺氧复氧后心肌细胞NFκB活性的升高。 Objective The present study is to investigate the effect of EPO pretreatment on TNF-α expression in cultured cardiac myocytes with H/R and to explore the possible NF-kB signal transduction mechanisan. Methods The model of cultured cardiac myocytes with H/R was established and the cardiac myocytes were divided into 4 groups, including EPO group （treat with EPO 10 U/ ml 24 h before H/R）, EPO ＋ PDTC group （treat with EPO 10 U/mL and PDTC 5μg/ml 24 h before H/R）, PDTC group （treat with PDTC 5μg/ml 24h before H/R） and control group. Change of TNF-α gene expression before and after H/R in cardiac myocytes was detected with RT-PCR and western blot. Change of NF-kB activity before and after H/R in cardiac myocytes was assayed with EMSA. Results Before H/R, there was no significant difference in TNF-α mRNA and protein expression between the 4 groups and after lad H/R TNF-α mRNA and protein expression increased signiticanfly in the 4 groups compared to eontrd group before H/R. After H/R, TNF-α mRNA and protein expression was lower in EPO group than in the other 3 groups. Before H/R, H/R activity was higher in EPO group than in the other groups. After H/R, NF-kB activity increased significantly in all the 4 groups compared to the control before H/R and NF-kB activity was lower in EPO group than in the other groups. groups EPO pretreatment inhibited the upregulation of TNF-α gene expression after H/R in cardiac myocytes, which might be reated to the inhibition of NF-kB activation; EPO pretreatment might inhibit the activation of NF-kB after H/R through the negstive feed-back mechanism of NF-kB activation.

作者秦川肖颖彬钟前进陈林王学锋

机构地区第三军医大学新桥医院心血管外科

出处《中华胸心血管外科杂志》 CSCD 北大核心 2006年第3期190-192,共3页 Chinese Journal of Thoracic and Cardiovascular Surgery

关键词心肌红细胞生成素肿瘤坏死因子 NF-kB 心肌再灌注损伤动物实验替代试验 Myocardium Eryghropoietin Tumor necrosis factor NF-kappa B Myocardial reperfusion injury Animal testing alternative

分类号 R541 [医药卫生—心血管疾病]

引文网络
相关文献

参考文献9

1Koyama T, Temma K, Akera T, et al. Reperfusion-induced contracture develops with a decreasing [ Ca^2+] : in single heart cells. Am J Physiol, 1991,261 : 159 - 163.
2Junk AK, Mammis A, Savitz Sl, et al. Erythropoietin administration protects retinal neurons from acute ischemia-reperfusion injury. Proc Natl Acad Sci U S A,2002,99:10659- 10664.
3Chong ZZ, Kang JQ, Maiese K. Erythropoietin is a novel vascular protectant throngh activation of Aktl and mitochondrial modulation of cysteine proteases. Circulation,2002,106:2973 - 2979.
4Juul S. Faythwpoiefin in the central nervous system, and its use to prevent hypoxic-ischemic brain damage. Acta Paediatr Suppl,2002,91:36-42.
5Parsa Cj, Matsumoto A, Kim J, et al. A novel protective effect of erythropoietin in the infarcted heart. J Clin Invest,2003,112:999- 1007.
6Moon C, Krawczyk M, Alan D, et al. Erythropoietin reduced myocardial infarction and left ventricular functional decline after coronary artery ligation in rats. Proc Nail Acad Sci USA,2003,100:11612- 11617.
7Calvillo L, Latini R, Kajstura J, et al. Recombinant human erythropoietin protects the myocardium from ischemia-reperfusion injury and promotes beneficial remodeling. Proc Natl Acad Sci USA,2003,100:4802- 4806.
8Ono K, Matsumofi A, Furukawa Y, et al. Prevention of myocardial reperfusion injury in rats by an antibody against monocyte chemotactic activating factor/monocyte chemoattractant protein-1. Lab Invest, 1999, 79:195 - 203.
9Morgan EN, Boyle EM, Yun W, et al. An essential role for NF-κB in the cardioadapfive response to ischemia. Ann Thorac Surg, 1999, 68:377 - 382.