穿心莲对人CD4+T细胞启动子CXCR4和CCR5的作用及机制

[Effect and mechanism of Andrographitis Herba on human CD4+ T cell promoters CXCR4 and CCR5].

作者信息

Feng Long, Zhao Guoqiang, Ma Yunyun, Li Min, Ma Jing, Jin Jing, Cui Ying

机构信息

Department of Pathogenic Organism Biology, Henan University of Traditonal Chinese Medicine, Zhengzhou 450008, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2011 Nov;36(21):3012-7.

PMID:22308693

Abstract

OBJECTIVE

Utilizing a gene reporter technique to study the effects of Andrographitis Herba on human CXCR4 and CCR5 promoters.

METHOD

Inhibition of CXCR4 and CCR5 on T cells of healthy volunteers was analyzed by RT PCR, Western blot and flow cytometry. The human CXCR4 and CCR5 promoters driving a luciferase reporter in vectors pGLA. 17-CXCR4 and pGLA. 17-CCR5 were transfected into H9 stem cells. G418 was used for selecting stable cell lines. Rat sera thus medicated was collected and added to the transfected H9 cells, in which the expression of CXCR4 and CCR5 promoters was detected.

RESULT

They showed that the mRNA and protein expression levels of CXCR4 and CCR5 in human CD4+ T cells decreased significantly after taking Andrographitis Herba (P<0.05). Furthermore human CXCR4 and CCR5 promoter activity was downregulated significantly by sera from rats medicated with Andrographitis Herba.

CONCLUSION

Andrographitis Herba may have the effect of down-regulating CXCR4 and CCR5 promoters. It provides a feasible experimental platform for screening herbal medicine as the treatment of HIV/AIDS.

摘要

目的

利用基因报告技术研究穿心莲对人CXCR4和CCR5启动子的影响。

方法

通过逆转录聚合酶链反应（RT PCR）、蛋白质免疫印迹法（Western blot）和流式细胞术分析穿心莲对健康志愿者T细胞中CXCR4和CCR5的抑制作用。将驱动荧光素酶报告基因的人CXCR4和CCR5启动子分别导入载体pGLA.17 - CXCR4和pGLA.17 - CCR5中，然后转染至H9干细胞。用G418筛选稳定细胞系。收集经穿心莲处理的大鼠血清并添加到转染后的H9细胞中，检测CXCR4和CCR5启动子的表达。