伴放线聚集杆菌细胞致死性膨胀毒素介导人T淋巴细胞凋亡的实验研究  

Investigation of the effect of aggregatibacter actinomycetemcomitans cytolethal distending toxin on inducing apoptosis in human Jurkat cells

作　　者：陈慧萍[1] 陈旭[1] 李璐[1,2] 孙颖[1,2] 徐艳[1,2] 

机构地区：[1]南京医科大学口腔疾病研究江苏省重点实验室,江苏南京210029 [2]南京医科大学附属口腔医院牙周科,江苏南京210029

出　　处：《口腔医学》2015年第5期365-369,共5页Stomatology

基　　金：国家自然科学基金(81170962;81470749);江苏高校优势学科建设工程资助项目(2014-37)

摘　　要：目的通过研究伴放线聚集杆菌(aggregatibacter actinomycetemcomitans,A.actinomycetemcomitans)的一种毒力因子细胞致死性膨胀毒素(cytolethal distending toxin,CDT)诱导人T淋巴细胞发生凋亡过程中表达改变的相关基因,探索其凋亡过程中可能存在的相关信号调节通路,寻找侵袭性牙周炎诊断、治疗潜在的生物标记分子。方法通过透射电镜和流式细胞仪检测细胞凋亡的情况,荧光定量PCR检测CDT蛋白作用于人T淋巴细胞后凋亡相关基因表达的改变。结果 CDT蛋白作用于人T淋巴细胞24 h后,处理组细胞凋亡比例约为29.1%,对照组约为6.3%,透射电镜观察到典型的凋亡细胞的形态,如细胞体积缩小、染色质浓缩及边集等现象。荧光定量PCR结果显示促凋亡基因GADD45A、TNFSF8、TRADD、TNFRSF10B和TP53表达升高。结论伴放线聚集杆菌CDT可以促进人T淋巴细胞的凋亡,在这一过程中相关的促凋亡基因表达增加。Objective 1） To investigate the apoptotic gene expression profile in T lymphocytes after aggregatibacter actinomycetem- comitans （A. actinomycetemcomitans） cytolethal distending toxin （CDT） infection, and to find out the molecules involved in this process. 2） To explore mechanisms of the development and progression of aggressive periodontitis （AgP） ,and to provide potential bio- markers for diagnosis and therapy. Methods After human Jurkat cells were treated by A. actinomycetemcomitans CDT for 24 hours,an- nexin V/PI detection andelectron microscopy technology was employed to detect the apoptosis. Apoptotic gene expression was examined by quantitative real-time PCR. Results By flow cytometry,the proportion of apoptotic cells was 29.1% in treatment groups,and that in the control group was 6. 3%. The typical features of apoptosis were observed under trans/nission electron microscopy in treatment groups,such as cell shrinkage, chromatin condensation, and marginalization. Five pro-apoptotic genes were upregulated, including GADD45A,TNFSF8,TRADD,TNFRSF10B and TP53. Conclusions A. actinomycetemcomitans CDT could promote apoptosis of hu- man T lymphocytes,and the expression of related pro-apoptotic genes increases during this process.

关 键 词：伴放线聚集杆菌 细胞致死性膨胀毒素 JURKAT细胞 凋亡 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]