金叶败毒制剂对人巨细胞病毒感染细胞病变及细胞周期影响.docVIP

金叶败毒制剂对人巨细胞病毒感染细胞病变及细胞周期影响 　【摘要】 　　目的探讨中药金叶败毒制剂（简称中药）对人巨细胞病毒（HCMV）感染人胚肺成纤维细胞（HEL）细胞病变及细胞周期的影响。方法用HCMV AD169感染人胚肺成纤维细胞，分别用金叶败毒制剂和更昔洛韦（简称GCV）干预后，用倒置相差显微镜观察不同感染时间细胞病变，并采用流式细胞技术检测HCMV感染及药物干预后宿主细胞周期。结果金叶败毒制剂能明显抑制HCMV感染HEL细胞病变。中药干预组在病毒感染48 h后，HEL细胞处于G2/M期的细胞明显增多,在HCMV感染72 h后，药物干预组 HEL细胞均表现为S期细胞明显减少，G2/M期和G0/G1期细胞明显增多。结论金叶败毒制剂可能通过延缓HCMV感染细胞病变，促进细胞周期进程而发挥抗病毒作用。 【关键词】 金叶败毒制剂； 人巨细胞病毒； 细胞病变； 细胞周期 　　Abstract：ObjectiveTo study the effect of Jinye Baidu Preparation (JBP)，a Chinese medicinal preparation, on the cell pathological change and cell cycle of human embryonic lung fibroblast cells(HEL) infected by human cytomegalovirus (HCMV). MethodsThe HEL cells were infected by HCMV AD169, the pathological change of the infected cells during different infected time was observed by inverted contrast microscope before and after intervention of JBP and GCV. The cell cycle was detected in the HEL and the infected cells before and after intervention of medicine by the Flow Cytometer. ResultsJBP showed obviously inhibitory action on cell pathological change in the HCMV infected cells. The infected cells in G2/M phase were obvious increased 48 hours after HCMV infection after intervention of JBP. The infected cells in S phase were obviously reduced, and in G2/M phase and in G0/G1 phase they were obviously increased 72 h after HCMV infection after intervention of JBP.ConclusionJBP can delay cell pathological change of the HCMV infected cells, advance the cell cycle process so as to achieve the anti-virus action. 　　Key words： Jinye Baidu Preparation；Human cytomegalovirus；Cell pathological change；Cell cycle 　　人巨细胞病毒（HCMV）是目前引起宫内感染的重要病原体之一，HCMV宫内传播与异常妊娠结局（流产、死胎、畸形、胎儿生长受限等）的发生关系密切，已成为影响胎儿发育的常见原因［1］。由于妊娠期HCMV感染尚无有效的治疗药物，本课题组在对金叶败毒制剂和巨细胞病毒感染的长期研究中发现，该中药制剂在体内外对HCMV均有明显的抑制作用［2，3］。本研究采用倒置相差显微镜动态观察感染细胞病变及流式细胞检测技术，探讨中药金叶败毒制剂对HCMV感染的人胚肺成纤维细胞（HEL）的细胞病变及细胞周期的影响，为临床用药提供实验依据。　 　　1 材料与仪器 　　1.1 药物金叶败毒制剂主要成分为金银花、鱼腥草、大青叶及蒲公英等，20 ml/支，每毫升含生药量1 g，批号：950430，由同济医院中西医结合研究所提供,临用时用细胞维持液（含3%DMEM）稀释成