ERK信号通路对病毒性心肌炎心肌细胞CAR表达的影响  

Roles of ERK Signal Transduction Pathway to Coxsackie-adenovirus Receptor(CAR) in Cardiocytes during Viral Myocarditis

作　　者：朱彬[1] 孙强[2] 张鹭鹭[1] 刘惊今[1] 姜苗苗[1] 王帆[1] 卢少玲[1] 孟玮[1] 白景玉 张烁[1] 

机构地区：[1]哈尔滨医科大学附属第二医院,黑龙江哈尔滨150086 [2]秦皇岛市第一医院,河北秦皇岛066000

出　　处：《现代生物医学进展》2015年第25期4828-4833,共6页Progress in Modern Biomedicine

基　　金：黑龙江省教育厅科学技术研究项目(115811176)

摘　　要：目的:观察细胞外信号调节激酶(Extracellular Regulated Protein Kinases,ERK1/2)信号通路对病毒性心肌炎(Viral Myocarditis,VMC)心肌细胞柯萨奇病毒-腺病毒受体(Coxsackie-adenovirus Receptor,CAR)表达的影响。方法:新生SD大鼠心肌细胞体外培养48 h后随机分为3组,除对照组外均体外接种柯萨奇B3m病毒(Coxsackievirus B,CVB),建立VMC细胞模型。C组:DMEM对照组;V组:CVB3m感染组;U+V组:接种病毒前30 min,给予ERK1/2通路抑制剂U0126(10μmol/L)。各组分别于种毒后12 h、24 h、36 h取心肌细胞,用Western blot法测定ERK1/2活化水平及CAR表达量,并按上述时间点观察各组心肌细胞形态、搏动情况、细胞损伤程度,取培养液测定乳酸脱氢酶(LDH)水平。结果:在接种病毒后12 h,V组与C组相比,P-ERK1/2表达增高(3.25±0.61 vs 0.59±0.09,P<0.05),CAR表达增高(1.03±0.17 vs 0.78±0.11,P>0.05),逐步出现细胞病变,细胞搏动停止,培养液中LDH水平明显增高(1016.67±67.75 vs 336.34±28.67,P<0.05),心肌酶学的升高与镜下心肌细胞损伤程度平行;U+V组与V组相比,P-ERK1/2表达降低(1.66±0.28 vs 3.25±0.61,P<0.05),CAR表达明显增高(1.73±0.27 vs 1.03±0.17,P<0.05),但细胞损伤却明显减轻,LDH水平明显降低(410.06±13.62 vs 1016.67±67.75,P<0.05)。动态观察24 h、36 h,同样出现上述变化趋势。结论:ERK1/2信号转导通路参与心肌细胞感染CVB发生急性损伤的过程,并参与调控CAR的表达。在病毒感染后36 h内,阻断ERK1/2信号通路,CAR表达上调,并未加重心肌细胞损伤。Objective： To investigate the role of extracellular signal-regulated kinase（ERK） signal transduction pathway to coxsackie-adenovirus receptor（CAR） in cardiocytes during viral myocarditis（VMC）. Methods： Primary cultured new born SD rat cardiocytes were infected by CVB3 m to establish a model of viral myocarditis. Forty-eight hours after culturing, cardiomyocytes were divided into 3 groups. Group C： cardiomyocytes were incubated with DMEM as controls; group V： cardiomyocytes were incubated with CVB solutions; group U＋V： cardiomyocytes were incubated with U0126（10 μmol/L） for 30 min initially, and then incubated with CVB solutions. After processing, the level of P-ERK1/2 and CAR were detected by Western blot. CVB mediated cytopathic effects were observed after co-culturing for further 12, 24, 36 hours respectively, lesions of the cardiocytes were examined by LDH assay. Results：Twelve hous after infected by CVB, comparing to group C, the level of P-ERK1/2 in group V increased（3.25±0.61 vs 0.59±0.09, P〈0.05）, the exrpession of CAR increased（1.03±0.17 vs 0.78±0.11, P〉0.05）, the beating rate decreased gradually and then stopped, the level of cardiac enzymes increased（1016.67 ±67.75 vs 336.34 ±28.67, P〈0.05）. In group U＋V, comparing to group V, the level of P-ERK1/2 decreased（1.66±0.28 vs 3.25±0.61, P〈0.05）, the exrpession of CAR increased significantly（1.73±0.27 vs 1.03±0.17, P〈0.05）, the injury to cardiocyte extenuates, so was the level of cardiac enzymes（410.06 ±13.62 vs 1016.67±67.75, P〈0.05）. During the following 24 and 36 hours, we can also observe the same change. Conclusions： When cardiocytes were infected by CVB3 m in acute phase, ERK1/2 signal transduction pathway was involved in it. It also participated in the regulation of CAR. Inhibition of this pathway did not aggravate the injury of cardiac cells.

关 键 词：病毒性心肌炎 ERK1/2 CAR 

分 类 号：R95-33[医药卫生—药学] R542.21