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醫(yī)學免費論文:B組柯薩奇病毒對心肌細胞感染模式的研究
【摘要】 目的 觀察B組柯薩奇病毒(Coxsackie virus B，CVB)在原代心肌細胞中的感染特點，從而研究CVB對心肌細胞的致病機制。方法 原代培養(yǎng)Balb/c乳鼠心肌細胞，并獲得純化的心肌細胞。使用B組柯薩奇病毒嗜心肌毒株3型(CVB3m)攻擊原代心肌細胞，觀察病毒感染心肌細胞后心肌細胞的細胞病變、超微結(jié)構(gòu)和心肌酶的變化。結(jié)果 成功原代培養(yǎng)了Balb/c乳鼠心肌細胞。心肌細胞在病毒感染后，細胞病變(CPE)不明顯，與在HeLa等細胞中的典型CPE不同。心肌細胞在CVB3m感染后保持細胞形態(tài)和博動超過2周時間。電鏡顯示心肌細胞結(jié)構(gòu)正常，細胞器數(shù)量增加，在心肌細胞胞質(zhì)中可見晶格樣病毒顆粒排列。培養(yǎng)細胞的上清液中心肌酶升高。結(jié)論 CVB在原代心肌細胞中表現(xiàn)為持續(xù)感染，CVB在心肌細胞中的致病機制不同于HeLa細胞。
【關(guān)鍵詞】 心肌細胞;B組柯薩奇病毒;Balb/c乳鼠;原代細胞培養(yǎng)
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.醫(yī).學全.在.線網(wǎng)站www.gydjdsj.org.cn
Key words: cardiomyocytes; Coxsackie virus B; sucking Balb/c mouse; primary cell culture
B組柯薩奇病毒(Coxsackievirus B，CVB)是病毒性心肌炎和心肌病的主要病因，其敏感動物模型是Balb/c乳鼠，如果要離體研究CVB對心肌的致病性，必須原代培養(yǎng)Balb/c乳鼠心肌細胞[1]。CVB感染細胞通常表現(xiàn)為急性殺細胞病變，例如，宮頸癌細胞HeLa細胞是CVB致病機制研究中最常用的細胞平臺，CVB感染HeLa細胞后，在24 h出現(xiàn)明顯細胞病變(CPE)，細胞變圓聚集，48 h細胞死亡脫落;理論上，CVB感染心肌細胞也應該表現(xiàn)為殺細胞效應。但是，分子流行病學研究證明，CVB可以持續(xù)存在心肌組織中。Quigley等[2]觀察23例臨床和組織學表現(xiàn)為心肌炎的患者，其中12例在平均43個月中發(fā)展為擴張型心肌病(DCM)，隨后4例死亡，2例心臟移植，1例痊愈，其余仍為DCM。本課題利用CVB1核酸、嗜心肌毒株B組柯薩奇病毒3型(CVB3m)攻擊原代培養(yǎng)的Balb/c乳鼠心肌細胞，觀察CVB在心肌細胞中的感染特點，從而說明CVB在心肌細胞中的感染機制。
