CVB3感染通过MCP-1介导病毒性心肌炎小鼠单个核细胞迁移

CVB3 infection induced migration of mononuclear cells of mice with viral myocarditis versus MCP-1

目的:研究B3型柯萨奇病毒(CVB3)感染与病毒性心肌炎(VMC)炎症细胞迁移的关系及机制。方法:采用差异贴壁法分离小鼠心肌细胞;趋化试验分析心肌细胞培养上清对VMC小鼠外周血单个核细胞(PMNC)的趋化性;实时定量RT PCR分析心肌细胞MCP 1的表达。结果:( 1)CVB3感染心肌细胞培养上清对VMC小鼠PMNC的趋化性明显增强。( 2 )CVB3感染2小时后MCP 1的表达开始升高,至6小时达到高峰,8小时下降;随着CVB3感染量的增加,MCP 1的表达明显升高。( 3)2 5 μg/ml抗MCP 1抗体处理后,CVB3感染心肌细胞培养上清对VMC小鼠PMNC的趋化性下降5 4 % (P <0 0 1) ;5 μg/ml抗MCP 1抗体处理后,趋化性下降的幅度与2 5 μg/ml抗MCP 1抗体处理的相比未见明显差异(P >0 0 5 )。结论:CVB3感染通过MCP 1介导VMC小鼠单个核细胞迁移。

Objective:To investigate relationship between coxsackievirus group B type 3(CVB3) infection and migration of inflammatory cells of mice with viral myocarditis(VMC) and its mechanism.Methods:Neonatal murine cardiac myocytes were isolated by different adhesion method.Chemoattractant of supernatant of primary cultured cardiac myocytes was detected to peripheral mononuclear cells(PMNC) of mice with VMC by chemtaxis assay.The expression of monocyte chemoattractant protein-1(MCP-1) in cultured cardiac myocytes was analyzed quantitatively by real-time RT-PCR.Results:(1)CVB3 infection enhanced the chemoattractant of supernatant of primary cultured cardiac myocytes to PMNC of mice with VMC.(2)The expression of MCP-1 was up-regulated at the second hour and climbed a peak at the 6th hour post infection in cardiac myocytes.However there was no significant difference within 8 hours in the expression of MCP-1 in uninfected cardiac myocytes.The expression of MCP-1 was up-regulated along with increase of CVB3 dose.(3)The chemoattractant of supernatant of cultured cardiac myocytes infected by CVB3 to PMNC of mice with VMC reduced 54% after pretreated with anti-MCP-1 antibody (2 5 μg/ml)( P <0 01).There was no significant difference in chemoattractant to PMNC of mice with VMC between 2 5 μg/ml anti-MCP-1 treated group and 5 μg/ml anti-MCP-1 antibody treated group ( P >0 05).Conclusion:CVB3 infection induced migration of mononuclear cells of mice with VMC versus MCP-1,which may be one of main mechanisms of infiltration of inflammatory cells in myocardium caused by CVB3 infection.