不同毒力结核分枝杆菌感染小鼠对肺泡巨噬细胞的凋亡率及其时相性变化的影响

Detection of apoptosis rate of alveolar macrophages in mice and change with the time for infection of various kinds of poisonous mycobacterium tuberculosis

目的:探讨不同毒力结核分枝杆菌感染小鼠肺泡巨噬细胞的凋亡率及其时相性变化。方法:不同毒力结核分枝杆菌悬液分别经小鼠尾静脉注射、复制及鉴定各组小鼠感染模型,各组小鼠感染模型复制成功后第1、3、5、7、9、11、13、15天,进行肺泡灌洗,收集小鼠肺泡灌洗液,获取感染小鼠肺泡巨噬细胞,以激光共聚焦显微镜技术检测及鉴定结核分枝杆菌感染小鼠肺泡巨噬细胞,流式细胞术检测上述各时间点、各组感染小鼠肺泡巨噬细胞的凋亡率,比较各组感染小鼠肺泡巨噬细胞凋亡率的时相性变化。结果:激光共聚焦显微镜检测结果显示,结核分枝杆菌国际标准强毒株H37Rv株和卡介苗菌株(BCG)均被小鼠肺泡巨噬细胞大量吞噬。流式细胞技术检测结果显示:结核分枝杆菌国际标准强毒株H37Rv株感染小鼠模型组和卡介苗菌株(BCG)感染小鼠模型组,在小鼠感染模型复制成功后1～9天,小鼠肺泡巨噬细胞的凋亡率逐渐升高,9天时两组小鼠肺泡巨噬细胞的凋亡率均达最高,随着时间的延长,两组小鼠肺泡巨噬细胞的凋亡率呈现逐渐降低趋势。结核分枝杆菌国际标准强毒株H37Rv株感染小鼠组,感染小鼠的肺泡巨噬细胞的凋亡率明显高于卡介苗菌株(BCG)感染小鼠组,差异有统计学意义(P<0.05)。结论:感染小鼠的肺泡巨噬细胞的凋亡率与结核分枝杆菌的毒力强弱呈正相关。

Objective：To analyze and study alveolar macrophages of mice＇ apoptosis rate and time phase change infected by various of poisonous mycobacterium tuberculosis.Methods：The mice were injected with mycobacterium tuberculosis bacteria suspension through the tail vein of mice to replicate models of mice infected with Mycobacterium tuberculosis.After the successful replication of the models,we cleaved the alveolar of every group of mice and collect cleavage liquid to obtain alveolar macrophages of the infected mice on the 1st,3rd,5th,7th,9th,11th,13th,15th days,then laser co focal microscopy was used to observe the model of alveolar macrophages of mice infected by mycobacterium tuberculosis.At the same time,macrophage＇ apoptosis rate was detected by flow cytometry and apoptosis rate of changing alveolar macrophages of every group was compared and analysed in different times.Results：The results of laser co focal microscopy showed that Mycobacterium tuberculosis H37Rv and BCG were swallowed largely by alveolar macrophage of mice.The results of flow cytometry showed that apoptosis rate of alveolar macrophages of the mice infected by Mycobacterium tuberculosis H37Rv and BCG gradually increased on the first 9 days and reached the maximum in the ninth day,with the time goes on,then gradually decreased.The apoptosis rate of alveolar macrophages of the mice infected by Mycobacterium tuberculosis H37Rv was higher than the BCG groups,the two groups was statistically significant（P0.05）.Conclusion：The apoptosis rate of alveolar macrophages of the mice is related to the virulence of Mycobacterium tuberculosis.