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医学免费论文:B组柯萨奇病毒对心肌细胞感染模式的研究
【摘要】 目的 观察B组柯萨奇病毒(Coxsackie virus B，CVB)在原代心肌细胞中的感染特点，从而研究CVB对心肌细胞的致病机制。方法 原代培养Balb/c乳鼠心肌细胞，并获得纯化的心肌细胞。使用B组柯萨奇病毒嗜心肌毒株3型(CVB3m)攻击原代心肌细胞，观察病毒感染心肌细胞后心肌细胞的细胞病变、超微结构和心肌酶的变化。结果 成功原代培养了Balb/c乳鼠心肌细胞。心肌细胞在病毒感染后，细胞病变(CPE)不明显，与在HeLa等细胞中的典型CPE不同。心肌细胞在CVB3m感染后保持细胞形态和博动超过2周时间。电镜显示心肌细胞结构正常，细胞器数量增加，在心肌细胞胞质中可见晶格样病毒颗粒排列。培养细胞的上清液中心肌酶升高。结论 CVB在原代心肌细胞中表现为持续感染，CVB在心肌细胞中的致病机制不同于HeLa细胞。
【关键词】 心肌细胞;B组柯萨奇病毒;Balb/c乳鼠;原代细胞培养
A study of the effect of infection modes of Coxsackie virus B on cardiomyocytes
ZHANG Zhonghai1, ZHANG Qi2, YAN Changdong1, ZHONG Zhaohua3
(1. Department of Physiology, Xuzhou Medical College, Xuzhou, Jiangsu 221004, China;
2. Laboratory Animal Center, Xuzhou Medical College; 3. Department of Microbiology,
Harbin Medical University, Harbin, Heilongjiang 150086)
Abstract: Objective To investigate the characteristics of Coxsackie virus B (CVB) on primarily cultured myocardial cells and to explore the pathogenesis of cardiomyocytes.Methods Cardiomyocytes from Balb/c suckling mouse were cultured in vitro, and purified myocardial cells were obtained. The cultured myocardial cells were attacked with CVB3m, a myocardial tropic Coxsackie virus B strain. The cytopathic effect (CPE) and ultrastructure of the CVB3m-infected myocardial cells were observed. The variations of myocardial enzymes in the supernant of the culture medium were determined.Results The cardiomyocytes from neonate Balb/c mice were successfully cultured. No apparent CPE could be observed in the cultured myocardial cells following the CVB3m attack, which was different from the typical CPE in the infections of HeLa cells. The cardiomyocytes retained intact morphology and pulsation for over two weeks as of the infection. The electronic microscopy showed that the myocardial cells had normal ultrastructure, with an increase in the number of organelles and visible crystally-arranged viral particles in the cytoplasm. The level of myocardial enzymes in the supernatant of the infected myocardial cells was higher than that of the control group.Conclusion CVB can persistently infect cardiomyocytes. The pathogenesis of CVB on myocardial cells differs from that in HeLa cell.医.学全.在.线网站www.med126.com
Key words: cardiomyocytes; Coxsackie virus B; sucking Balb/c mouse; primary cell culture
B组柯萨奇病毒(Coxsackievirus B，CVB)是病毒性心肌炎和心肌病的主要病因，其敏感动物模型是Balb/c乳鼠，如果要离体研究CVB对心肌的致病性，必须原代培养Balb/c乳鼠心肌细胞[1]。CVB感染细胞通常表现为急性杀细胞病变，例如，宫颈癌细胞HeLa细胞是CVB致病机制研究中最常用的细胞平台，CVB感染HeLa细胞后，在24 h出现明显细胞病变(CPE)，细胞变圆聚集，48 h细胞死亡脱落;理论上，CVB感染心肌细胞也应该表现为杀细胞效应。但是，分子流行病学研究证明，CVB可以持续存在心肌组织中。Quigley等[2]观察23例临床和组织学表现为心肌炎的患者，其中12例在平均43个月中发展为扩张型心肌病(DCM)，随后4例死亡，2例心脏移植，1例痊愈，其余仍为DCM。本课题利用CVB1核酸、嗜心肌毒株B组柯萨奇病毒3型(CVB3m)攻击原代培养的Balb/c乳鼠心肌细胞，观察CVB在心肌细胞中的感染特点，从而说明CVB在心肌细胞中的感染机制。
