姜黄素对HIV-1辅助受体CCR5启动子的干预作用  

Effect of curcumin on HIV-1 co-receptor CCR5 promoter

作　　者：李志慧 冯龙(指导)[1] 郑文锦 孙颖[1] 夏金婵[1] 梅雪[1] 江华[1] 李晓娟[1] LI Zhi-Hui;FENG Long;ZHENG Wen-Jin;SUN Ying;XIA Jin-Chan;MEI Xue;JIANG Hua;LI Xiao-Juan(College of Medicine,Henan University of Traditional Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学医学院,郑州450046

出　　处：《中国免疫学杂志》2021年第16期1953-1957,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金项目(81503677);河南省高等学校重点科研项目(20A310008)。

摘　　要：目的:探讨姜黄素对HIV-1辅助受体CCR5启动子活性的影响。方法:构建pFireRluc-CCR5重组载体,瞬时转染293T细胞;采用CCK-8法检测姜黄素对293T细胞活性的影响;利用双荧光素酶报告基因技术检测姜黄素对CCR5启动子活性的影响。结果:PCR方法分别扩增出CCR5启动子(987 bp)和Rluc(1 997 bp)表达单元,双荧光素酶报告基因表达载体pFireRluc-CCR5经酶切和测序鉴定,插入正确;根据CCK-8实验结果确定姜黄素给药浓度分别为10、20、40、60、80μmol/L;加药后,CCR5启动子活性显著下降(P<0.05),推测姜黄素能够显著降低CCR5启动子活性。结论:成功构建了pFireRluc-CCR5双荧光素酶报告基因表达载体,并证实其活性,姜黄素对CCR5启动子活性有显著抑制作用,具有潜在的抗HIV-1作用。Objective:To investigate the effect of curcumin on HIV-1 co-receptor CCR5 promoter activity.Methods:Construct pFireRluc-CCR5 recombinant vector and transiently transfect 293 T cells. CCK-8 cytotoxicity test was used to detect the effect of curcumin on 293 T cell activity. Dual luciferase reporter gene technology was used to detect the effect of curcumin on CCR5 promoter activity.Results:PCR method was used to amplify the CCR5 promoter(987 bp)and Rluc(1 997 bp)expression units,the dual luciferase reporter gene expression vector pFireRluc-CCR5 was identified by digestion and sequencing. According to the results of CCK-8 experiment,curcumin was administered at concentrations of 10,20,40,60,80 μmol/L. After adding medicine,the activity of CCR5 promoter was significantly down-regulated(P<0.05). It was speculated that curcumin could significantly reduce the activity of CCR5 promoter.Conclusion:The pFireRluc-CCR5 dual luciferase reporter gene expression vector is successfully constructed and its activity is confirmed. Curcumin has a significant inhibitory effect on the activity of the CCR5 promoter and has a potential anti-HIV-1 effect.

关 键 词：姜黄素 CCR5启动子 HIV-1 载体构建 

分 类 号：R392[医药卫生—免疫学]