小鼠病毒性心肌损伤模型制作及cTnI基因扩增与测序分析  

Preparation of mouse viral cardimyocyte injury model and amplification and sequencing of myocardial troponin I gene

作　　者：邵义祥[1] 姚登福[2] 朱健华[2] 高增栋[2] 吴玮[2] 吴信华[2] 邱历伟[2] 

机构地区：[1]南通医学院实验动物中心 [2]南通医学院附属医院

出　　处：《临床心血管病杂志》2005年第1期29-32,共4页Journal of Clinical Cardiology

基　　金：江苏省自然科学基金项目资助(No:BK99160)

摘　　要：目的:分析心肌型肌钙蛋白(cTnI) mRNA在监测病毒性心肌损伤发生与发展过程中的价值。方 法:于BALB/c小鼠腹腔接种1×108TCID50柯萨奇病毒B3(CVB3)诱发心肌损伤,分别在CVB3感染后第3、6、9、 12、15、18和21天采集外周血后处死小鼠,留取鼠心脏作病理组织学检查,以逆转录 聚合酶链反应(RT PCR)分 析心肌和外周血中cTnI基因及测序分析扩增片段。结果:CVB3感染后,鼠心肌组织及循环血中cTnI mRNA均 表达增加,且与心肌细胞肿胀、炎性细胞浸润、核固缩及碎裂、变性、坏死、钙化等组织学改变相关。cTnI mRNA 基因扩增,心肌组织全数阳性;外周血阳性率在对照组及感染后的第3、6、9、12、15、18和21天分别为0、0、0、 16.7%、40%、71.4%、83.3%和87.5%。测序分析证实从心肌组织及外周血中扩增基因片段与原序列完全一致。 结论:病毒性心肌损伤时cTnI mRNA上调表达并释放入血,循环血cTnI mRNA为监测心肌损伤发生与发展的 灵敏标志物。Objective: To investigate the value of circulating cTnI-mRNA detection for monitoring myocardial injury and development. Method: Viral myocardial injury models in BALB/c mice were created by intraperitoneal inoculation with coxsackievirus B3 (CVB 3, 1×10 8 TCID 50) . Total RNAs were extracted and cTnI-mRNA in mice cardiac tissues and circulating blood were amplified by nested-PCR during mice myocardial injury.Result: In viral infected mice, the mRNA abundance for cTnI was upregulated in heart and circulating blood and associated with salient myocardial histopathologic features, including myocardial swell, inflammatory cell infiltration, pyknosis, karyorrhexis, karyolysis, denaturalization, necrosis, and calcification. The cTnI-mRNA from infected-mice heart and circulating blood cells were analyzed by RT-PCR, the gene amplified positive fragment were found in all heart tissues. The incidence of cTnI-mRNA was 0, 0, 0, 16.7%, 40%, 71.4%, 83.3%, and 87.5% in the controls, the 3rd, 6th, 9th, 12th, 15th, 18th, and 21st day in circulating bloods from the infected mice, respectively. the PCR products from human heart tissues or peripheral blood were confirmed by sequencing analysis. Conclusion:The present data suggest that cTnI-mRNA expression is upregulated and released into blood on viral myocardial injury, and it is a sensitivity marker for monitoring myocardial injury and development.

关 键 词：柯萨奇病毒B组 心肌损伤 肌钙蛋白Ⅰ 基因表达 

分 类 号：R373[医药卫生—病原生物学]