用乳鼠动物模型进行轮状病毒感染肠外损害的试验-第三军医大学学报.DOC

轮状病毒感染致乳鼠肠外损害的实验研究 陈军华 解元元 朱朝敏 刘作义 (重庆医科大学附属儿童医院感染消化科，重庆400014) 提 要：目的 了解轮状病毒（Rotavirus, RV）感染引起肠外脏器损害的情况并探索其发病机制。方法 用MA104细胞培养猴RV SA-11经口感染生后7 d昆明小鼠乳鼠，实验组每只接种0.1 ml培养物冻融液（3.75×107 PFU/ml），对照组经口接种0.1 ml正常对照细胞冻融液。接种后3 d处死乳鼠，光镜下观察脑、肺、心、肝、胰和肾组织病理改变，电镜下观察心肌细胞超微结构改变；以免疫荧光方法检测各器官RV抗原分布情况；以Phalloidin-TRITC对丝状肌动蛋白染色并经软件（Imageproplus5.1, IPP5.1）测量分析以观察心肌组织丝状肌动蛋白含量有无改变；以原位凋亡细胞检测方法检测肝细胞和心肌细胞凋亡情况；以巢式RT-PCR方法检测各器官内RV RNA分布情况。结果 在光镜下脑、肺、胰和肾组织未见明显病变，但可见心肌间质水肿，心肌细胞浊肿，肝细胞空泡变性；电镜下心肌细胞肌丝溶解，线粒体内室扩张，滑面内质网扩张，核周间隙增宽；Phalloidin-TRITC染色未见心肌组织丝状肌动蛋白含量改变；除脑组织外其他器官内见到RV抗原分布，而各器官内均有RV-RNA分布；肝细胞凋亡增加，而心肌细胞未见凋亡。结论 RV可播散到肠外脏器并引起病理损害，但总体上程度较轻。病毒感染导致丝状肌动蛋白解聚及细胞骨架损害、诱导细胞凋亡不仅是肠粘膜损伤的机制，同时也是肠外损害的机制。 关键词：轮状病毒 感染 乳鼠 肠外损害 Extraintestinal lesions caused by rotavirus: An experimental study on suckling mouse Chen Jun-Hua, Zhu Chao-Min, Xie Yuan-Yuan, Liu Zuo-Yi*　(Department of Infection, Childrens Hospital, Chongqing University of Medical Sciences, Chongqing 400014, China） Abstraction: Objective To study the extraintestional lesions induced by rotavirus (RV) infection and explore the pathogenesis. Methods Simian rotavirus SA11 was cultured in MA-104 cells. After inoculation, the pathological changes in brain, lung, heart, liver, pancreas and kidney were observed, the RV antigens was detected, and the apoptotic cells were observed. Besides, we stained the filamentous actin(F-actin) with Phalloidine-TRITC, and then quantified the F-actin amount. Results Several pathological changes were found including myocardium tissue interstitial edema, cardiocyte granular degeneration, hepatic congestion, hepatocellular vacuolar degeneration. Such changes were not found in brain, lung, and pancreas. Meanwhile, several ultrastructural changes were found including dissolving myocardial F-actin, extending smooth endoplasmic reticulum, swelling mitochondria, and widening perinuclear space. No differences were seen in the quantity of myocardial F-actin. Apoptosis was found in liver cells, but not in myocardial cells. In the