H9c2细胞对B组柯萨奇病毒3型重组株的易感性

林乐勋1,2; 赵文然3; 武帅钦2; 佟雷2; 王燕2; 张凤民2; 钟照华1,2

微生物与感染 ›› 2011, Vol. 6 ›› Issue (2) : 78-83.

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微生物与感染 ›› 2011, Vol. 6 ›› Issue (2) : 78-83.
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H9c2细胞对B组柯萨奇病毒3型重组株的易感性

  • 林乐勋1,2; 赵文然3; 武帅钦2; 佟雷2; 王燕2; 张凤民2; 钟照华1,2
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Infectivity of Recombinant Coxsackievirus B3 in H9c2 cells

  • LIN Le-Xun1,2; ZHAO Wen-Ran3; WU Shuai-Qin2; TONG Lei2; WANG Yan2; ZHANG Feng-Min2; ZHONG Zhao-Hua1,2
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摘要

H9c2细胞是来源于大鼠胚胎心脏组织的成肌细胞系,B组柯萨奇病毒(coxsackievirus B,CVB)是心肌炎和扩张型心肌病的主要病原。本研究观察了B组柯萨奇病毒3型(CVB3)在H9c2细胞中的感染性,探讨H9c2细胞是否可用于CVB致心肌疾病的实验研究。用整合了增强型绿色荧光蛋白(EGFP)或海肾荧光素酶(RLuc)的CVB3重组株EGFP-CVB3、RLuc-CVB3攻击H9c2细胞和大、小鼠原代心肌细胞和骨骼肌细胞,应用荧光显微镜、流式细胞术和化学发光技术观察感染细胞的EGFP和RLuc表达,应用反转录-聚合酶链反应(RT-PCR)方法检测病毒在核酸水平的表达。结果显示,在病毒攻击的H9c2细胞未检测到EGFP和RLuc表达;RT-PCR显示在CVB3感染9 d后检测不到CVB3 RNA;CVB3毒株可感染小鼠心肌细胞、骨骼肌细胞和大鼠心肌细胞,但不感染大鼠骨骼肌细胞和H9c2细胞。本研究结果表明,H9c2细胞不是CVB3易感细胞,不能用于CVB3致心肌疾病的研究;该结果从病毒感染方面也支持H9c2具有大鼠骨骼肌细胞表型。

Abstract

H9c2 cell is a cardiac myoblastic cell line derived from rat heart tissue. Group B coxsackievirus (CVB) is the major pathogen of viral myocarditis and dilated cardiomyopathy. In this study, infectivity of CVB3 in H9c2 cells was observed to evaluate the value of H9c2 cells in the experimental study of CVB-induced myocardial diseases. H9c2 cells and cardiac and skeletal myocytes of Balb/c mouse and Sprague Dawley (SD) rat were infected with two CVB3 variants integrated with enhanced green fluorescence (EGFP) or Renilla luciferase (RLuc). The expressions of EGFP and RLuc were measured by fluorescence microscopy, flow cytometry, and chemiluminescence assays, and the viral genomic RNA level was detected by reverse transcriptase-polimerase chain reaction (RT-PCR). The results showed that neither EGFP expression nor RLuc expression could be detected in H9c2 cells infected with EGFP-CVB3 or RLuc-CVB3. The viral RNA could not be detected by RT-PCR from day 9 post-infection. The cardiac and skeletal myocytes of Balb/c mouse and cardiac myocytes of SD rat could be infected by CVB3, but the skeletal myocytes of SD rat and H9c2 cells could not. Taken together, this study suggests that H9c2 cell is not permissive cell for CVB3, and is not suitable for CVB-induced myocardial disease study. The results also support that H9c2 cells exhibit properties of skeletal myocytes from the view of infection.

关键词

H9c2细胞 / B组柯萨奇病毒3型 / 心肌细胞 / 骨骼肌细胞 /

Key words

H9c2 cell / Coxsackievirus B3 / Cardiomyocyte / Skeletal muscle cell / Murine

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林乐勋1,2; 赵文然3; 武帅钦2; 佟雷2; 王燕2; 张凤民2; 钟照华1,2. H9c2细胞对B组柯萨奇病毒3型重组株的易感性[J]. 微生物与感染. 2011, 6(2): 78-83
LIN Le-Xun1,2; ZHAO Wen-Ran3; WU Shuai-Qin2; TONG Lei2; WANG Yan2; ZHANG Feng-Min2; ZHONG Zhao-Hua1,2. Infectivity of Recombinant Coxsackievirus B3 in H9c2 cells[J]. Journal of Microbes and Infections. 2011, 6(2): 78-83

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