四嗪二甲酰胺抑制T淋巴细胞活化的实验研究  

Experimental study of inhibition of ZGDHu-1 on T lymphocytes activation

作　　者：邱莲女[1] 周永列[1] 陈秀君[2] 胡惟孝[3] 

机构地区：[1]浙江省人民医院检验中心,杭州310014 [2]温州医学院 [3]浙江工业大学药学院

出　　处：《中华微生物学和免疫学杂志》2010年第2期110-114,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目（30973568）;浙江省医学重点学科建设基金（07-010）

摘　　要：目的研究四嗪二甲酰胺（ZGDHu-1）对植物血凝素（PHA）刺激的T淋巴细胞体外活化的影响，初步探讨其免疫抑制作用。方法从健康人外周血分离淋巴细胞，经PHA刺激诱导淋巴细胞活化，ZGDHu-1单独或联合环孢霉素A（CSA）作用24h、48h，检测各组T淋巴细胞的增殖、凋亡情况。采用流式细胞仪检测CD3^＋CD69^＋、CD3^＋CD25^＋、CD4^＋CD25^＋、CD8^＋CD25^＋和CD3^＋Fas^＋、CD4^＋Fas^＋、CD8^＋Fas^＋。用Annexin V／PI染色结合流式细胞术，分析淋巴细胞早期凋亡。酶联免疫吸附法（ELISA）检测细胞培养上清IL-2和转化生长因子-β1（TGF-β1）。结果ZGDHu-1能明显降低PHA活化的CD3^＋CD69^＋、CD3^＋CD25^＋、CD4^＋CD25^＋和CD3^＋Fas^＋、CD4^＋Fas^＋、Annexin V^＋／PI^-，并降低活化T淋巴细胞的IL-2分泌及促进TGF—β1的分泌，联合CSA应用效果更明显。结论ZGDHu—1能抑制T淋巴细胞活化，降低T淋巴细胞凋亡，和CSA联合具有一定的协同效应。Objective To investigate the effects of ZGDHu-1 on T lymphocytes activation in vitro to elucidate its immanosuppressive effects. Methods Lymphocytes isolated from healthy persons were stimulated with phytohaemagglutinin（PHA） and different experimental groups were set by cocuhured for 24 h, 48 h with ZGDHu-1 or with ZGDHu-1 and Cyelosporin A（ CSA ）. To assess the proliferation and apoptosis of T lymphocytes, we detected CD3^＋ CD69^＋ , CD3^＋ CD25^＋ , CD4^＋ CD25^＋ , CD8^＋ CD25 ＋ and CD3^＋ Fas^＋ , CD4^＋ Fas^＋ , CD8 ＋ Fas^＋ with flow cytometry. The early apoptosis rate of lymphocytes was analyzed with flow cytometry. Culture supernatant IL-2 and TGF-β1 were detected with ELISA. Results ZGDHu-1 decreased PHA activative CD3^＋ CD69^＋ , CD3^＋ CD25^＋ , CD4^＋ CD25^＋ and CD3^＋ Fas^ ＋ , CD4^＋ Fas^ ＋ , Annexln V ^＋/ PI^- and inhibited IL-2 secretion and promoted TGF-β1 secretion respectively. ZGDHu-1 has synergistic effect with CSA to be more obvious. Conclusion ZGDHu-1 can inhibit T lymphocytes activation and decreased apoptosis of T lymphocytes. ZGDHu-1 has synergistic effect with CSA to be obvious.

关 键 词：ZGDHu-1 环孢霉素A T细胞活化 细胞凋亡 流式细胞术 

分 类 号：R392.1[医药卫生—免疫学]