染料木黄酮抑制HCMV感染人星形胶质细胞的机制  

MECHANISM OF GENISTEIN INHIBITION OF HUMAN CYTOMEGALOVIRUS INFECTION IN HUMAN ASTROCYTES

作　　者：徐敬国[1] 李玲[1] 于向民[1] 魏艳[1] 王斌[2] 钱冬萌[2] 

机构地区：[1]青岛大学医学院组织学与胚胎学教研室,山东青岛266071 [2]青岛大学医学院病原微生物学教研室,山东青岛266071

出　　处：《青岛大学医学院学报》2016年第3期260-264,共5页Acta Academiae Medicinae Qingdao Universitatis

基　　金：国家自然科学基金资助项目(81070501);山东省科技发展计划项目(2011YD18005);青岛市博士后应用研究项目(2015153)

摘　　要：目的探讨染料木黄酮（Gen）抑制人巨细胞病毒（HCMV）感染人星形胶质细胞的效果及机制。方法实验设立空白对照组（C组）、Gen对照组（G组）、HCMV组（V组）、HCMV＋Gen组（V＋G组）及Gen＋HCMV组（G＋V组）。采用CCK-8筛选合适的药物浓度。采用反转录-实时定量PCR技术检测各组病毒即刻早期蛋白编码基因IE1和IE2、宿主hsa-miR-200家族及病毒miR-UL112的表达;采用染色质免疫共沉淀法检测与HCMV主要即刻早期启动子（MIEP）结合的组蛋白H3乙酰化的变化。结果筛选出实验用药物浓度为10μmol/L。与V组相比,G＋V组和V＋G组IE1、IE2基因的表达下调（F=14.790~126.000,P〈0.05）,在部分时间段内hsa-miR-200家族及miR-UL112表达上调（F=3.041~4 638.724,P〈0.05）,与病毒MIEP结合的组蛋白H3的乙酰化降低（F=18.184~32.848,P〈0.05）。结论 Gen能一定程度抑制HCMV感染人星形胶质细胞所致的增殖异常,其机制可能为Gen能一定程度抑制IE1和IE2基因的表达以及与病毒MIEP结合的组蛋白H3的乙酰化,并上调部分宿主hsa-miR-200microRNA及病毒miR-UL112的表达。Objective To explore the effectiveness and its mechanism of Genistein（Gen）in inhibition of human cytomegalovirus（HCMV）infection in human astrocytes（AS）. Methods Five groups were set in this study：blank control group（group C）,Gen-control group（group G）,HCMV group（group V）,HCMV＋Gen group（group V＋G）and Gen＋HCMV group（group G＋V）.CCK-8assay was used to screen out the appropriate concentration of Gen.RT-qPCR was used to detect the expression changes of viral encoding gene IE1 and IE2protein,host＇s hsa-miR-200 miRNA cluster and viral miR-UL112.Chromatin immunoprecipitation assay（ChIP）was used to detect the changes of histone H3 acetylation binding major immediate early promoter（MIEP）. Results 10μmol/L of Gen was screened out to be used as the experimental drug concentration.Compared with group V,the expressions of IE1 and IE2genes down-regulated（F=14.790-126.000,P〈0.05）,and in some time periods,the expressions of hsa-miR-200 miRNA cluster and miR-UL112up-regulated（F=3.041-4 638.724,P〈0.05）,and the acetylation of histone H3 binding to viral MIEP declined（F=18.184-32.848,P〈0.05）. Conclusion Genistein can,to some extent,inhibit the abnormal proliferation of human astrocytes caused by HCMV infection.Its mechanism is likely that Genistein can inhibit the expressions of IE1 and IE2gene to a certain degree and the acetylation of histone H3 binding to viral MIEP and partly up-regulate the expressions of host＇s hsa-miR-200 miRNA cluster and viral miR-UL112.

关 键 词：巨细胞病毒 小神经胶质细胞 染料木黄酮 微RNAS 免疫沉淀法 

分 类 号：R511[医药卫生—内科学]