染料木黄酮对HCMV感染的人星形胶质细胞即刻早期蛋白表达影响  

INFLUENCE OF GENISTEIN ON IMMEDIATE EARLY PROTEIN EXPRESSION IN HUMAN ASTROCYTES INFECTED WITH HUMAN CYTOMEGALOVIRUS

作　　者：张志超[1] 李玲[1] 于向民[1] 卢桂玲[2] 王斌[3] 钱冬萌[3] 

机构地区：[1]青岛大学医学院组织胚胎学教研室,山东青岛266071 [2]青岛市市立医院高压氧科 [3]青岛大学医学院病原微生物学教研室

出　　处：《青岛大学医学院学报》2015年第6期631-633,637,共4页Acta Academiae Medicinae Qingdao Universitatis

基　　金：国家自然科学基金资助项目(81070501)

摘　　要：目的探讨染料木黄酮（Gen）对人巨细胞病毒（HCMV）感染的人星形胶质细胞（AS）即刻早期蛋白72（IE72）表达的影响。方法选取对数生长期的AS进行以下实验。HCMV组：HCMV感染AS;Gen组：用含40μmol/L的Gen与含体积分数0.02的FBS的DMEM/F12培养液培养AS;Gen＋HCMV组：用含40μmol/L的Gen与含体积分数0.02FBS的DMEM/F12培养液预培养AS 4h后,HCMV感染AS;对照组：仅以含体积分数0.02FBS的DMEM/F12培养液培养AS。分别用噻唑蓝（MTT）法检测Gen对AS增殖的影响,Real-time PCR和Western blot方法检测IE72mRNA和蛋白的表达。结果 MTT法检测结果显示,Gen＋HCMV组24、48、72h的吸光度值均高于HCMV组（t=5.94～12.99,P〈0.05）。48h时,HCMV组细胞有明显的细胞病变效应（CPE）,而Gen＋HCMV组细胞状态良好,CPE不明显。Western blot方法检测结果显示,Gen＋HCMV组IE72蛋白表达水平低于HCMV组（t=4.90～7.67,P〈0.05）。Real-time PCR结果显示,感染24、48、72和96h时,Gen＋HCMV组IE72mRNA的表达低于HCMV组（t=9.70～18.61,P〈0.05）。结论适当浓度的Gen能抑制HCMV感染所致的AS异常增殖,并抑制IE72蛋白的表达。Objective To explore the effect of Genistein（Gen）on immediate-early protein（IE72）expression in human Astrocytes（AS）infected by human cytomegalovirus（HCMV）. Methods Experiments of logarithmic growth phase AS were done as follows：HCMV group：AS infected with HCMV;Gen group：AS was cultivated with culture solution containing 40μmol/L of Gen and volume fraction of 0.02 FBS DMEM/F12;Gen＋HCMV group：AS was cultivated with the solution same as used in the Gen group for 4hours and then infected with HCMV;control group：AS cultivated with solution containing volume fraction of0.02 FBS DMEM/F12 only.The effect of Gen on AS proliferation was detected with MTT,and the expressions of IE72 mRNA and protein were measured using Real-time PCR and Western blotting. Results The results of MTT method showed that：in Gen＋HCMV group,the 24-,48-and 72-hour absorbance values were higher than HCMV group（t=5.94-12.99,P〈0.05）.At48 hours after infection,the cells in HCMV group showed obviously cytopathic effects（CPE）,and the cells in Gen＋HCMV group were in good condition,no obvious CPE was observed.Western blotting showed the expression of IE72 in Gen＋HCMV group was lower than that in HCMV group（t=4.90-7.67,P〈0.05）.Real-time PCR indicated that at 24,48,72 and 96hours of infection,the expression of IE72 mRNA in Gen＋HCMV group was lower than that in HCMV group（t=9.70-18.61,P〈0.05）. Conclusion Appropriate concentration of Genistein can inhibit abnormal proliferation of Astrocytes caused by infection of human cytomegalovi-rus and expression of IE72 mRNA.

关 键 词：染料木黄酮 巨细胞病毒 即刻早期蛋白 星形胶质细胞 

分 类 号：R285[医药卫生—中药学]