柯萨奇B3m病毒致小鼠急性心肌损伤的双向凝胶电泳分析

Analysis of acute myocardium injury animal model by Coxsackie virus B3m by two-dimensional electrophoresis

目的建立双向凝胶电泳体系分析柯萨奇B3m病毒(CVB3m)致小鼠急性心肌损伤过程中的蛋白质组差异,探讨CVB3m致心肌损伤的机制。方法分别提取感染CVB3m第0,4,8及21天的BALB/C鼠心肌组织总蛋白,采取固相pH梯度双向凝胶电泳技术分离总蛋白,用Image Master软件进行图像分析。结果心肌组织总蛋白平均抽提浓度为13.9 g/L;获得正常小鼠和急性CVB3m感染鼠不同感染时期(第4,8和21天)心肌组织双向电泳凝胶图谱,图像分析4块胶平均蛋白点数为(792±32)个,匹配率为61.25%±6.25%。随机选定不同感染时期的8个差异表达的蛋白质点,得到了表观相对分子质量和表观等电点等一系列数据。结论获得了CVB3m致小鼠急性心肌损伤的双向凝胶电泳图谱,建立了该模型的蛋白质双向凝胶电泳数据库,有利于该病毒引起小鼠心肌损伤机制的进一步研究。

Objective In order to discover the mechanism of acute myocardium injury by Coxsackie virus B3m(CVB3m), this study is designed to establish two-dimensional electrophoresis system to analyze proteome of acute myocardium injury animal model by CVB3m. Methods Extract protein of heart tissue from mice which were killed at 0, 4, 8, 21 days after virus injected. Then a series of methods were used including immobilized pH gradient two-dimensional electrophoresis,sliver staining, image master two-dimensional electrophoresis analysis software. Results The average protein consistency was 13.9 g/L. We got two-dimensional electrophoresis image of heart tissue from normal mice and infected mice which were killed at 4, 8, 21 days after virus injected. The average detected spot number was 792±32, the matching rate was 61.5%±6.25%. Eight proteins were found with the data of pI and Mr which had different volumes at different day after virus infected. Conclusions The data we got from this study will be useful for the discovery of mechanism of myocardium injury by CVB3m and the establishment of acute injury myocardium animal model by CVB3m proteome database.