鼠巨噬细胞Ana-1培养弓形虫RH株速殖子的特性研究  

Characteristics of cultivation of Toxoplasma gondii RH strain tachyzoites in murine Ana-1 macrophages

作　　者：郝永新[1] 李雪莲[1] 刘群[1] 丁隽[1] 张维[1] 

机构地区：[1]中国农业大学动物医学院,北京100193

出　　处：《中国兽医科学》2009年第3期196-203,共8页Chinese Veterinary Science

基　　金：农业部公益项目(200803017);国家自然科学基金项目(38071861);北京市自然科学基金项目(6082014)

摘　　要：为比较鼠巨噬细胞Ana-1（吞噬细胞）和Vero细胞（非吞噬细胞）培养弓形虫RH株后虫体以及虫体和宿主细胞间的表现，采用一般形态学观察、流式细胞术检测弓形虫接种后Ana-1细胞和Vero细胞的凋亡和坏死情况，并用NO检测试剂盒检测Ana-1细胞的NO分泌量。结果显示，弓形虫能够在两种细胞上大量发育和繁殖，而且两种细胞凋亡和坏死并存，Ana-1细胞以坏死为主（坏死率为58.6％，凋亡率为12.6%），Vero细胞则以凋亡为主（坏死率为28.7％，凋亡率为56.0％）。对NO的检测显示，含100mL／L小牛血清培养的Ana-1 NO分泌浓度为0.16-0.17μmol／mL；而感染弓形虫速殖子的Ana-1 NO分泌浓度为0.18-0.46μmol／mL，两者差异显著（P〈0.05）。证实，应用Ana-1和Vero细胞培养弓形虫均可获得大量虫体；弓形虫感染后吞噬细胞和非吞噬细胞的凋亡率和坏死率不同，差异极显著（P〈0.01）；弓形虫感染能够诱导Ana-1分泌NO。In order to compare the characteristics of cultivation of Toxoplasma gondii RH strain tachyzoites in murine Ana-1 macrophages （phagocytic cells） to those in Vero cells（non-phagocytic cells）, the apoptosis and necrosis of the Ana-1 cells and Vero cells induced by T. gondii were determined by morphological observation and the fluorescence activated cell sorting（FACS） assay,and NO secretion of the Ana-1 cells was determined by the NO molecular kit. The FACS data showed that the necrosis rate and apoptosis rate in the Ana-1 cells were 58.6% and 12.6%,respectivety,and the necrosis rate and apoptosis rate in the Vero cells were 28.7% and 56.0% ,respectively. The secretions of NO of the Ana-1 cells were 0.18-0.46μmol/mL and 0.16-0.17μmol/mL induced by calf serum and T. gondii（P〈0.05）,respec tively. These results indicated that large quantities of T.gondii could be obtained by cultivating in murine Ana-1 macrophages and Vero cells. Different proportions of apoptosis and necrosis were observed in the phagocytic cells and the non phagocytic cells（P〈0.01）. T. gondii infection could induce Ana-1 cells to secrete NO.

关 键 词：弓形虫 Ana-1细胞 流式细胞术 细胞凋亡 坏死 

分 类 号：S852.723[农业科学—基础兽医学]