新型融合多肽HTPP-MDC体外抑制HBV复制活性及亚细胞定位  

HTPP-MDC inhibits replication and subcellular localization of hepatitis B virus in HepG2.2.15 cells

作　　者：卢雪梅[1,2] 黄演婷[1] 汪洁[1] 金小宝[1] 朱家勇[1] 

机构地区：[1]广东药学院药用生物活性物质研究所,广东省生物活性药物研究重点实验室,广东广州510006 [2]南方医科大学公共卫生与热带医学学院,广东广州510515

出　　处：《中国病理生理杂志》2013年第7期1283-1287,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30671832);广东省医学科研基金资助项目(No.A2012299)

摘　　要：目的:研究肝细胞靶向穿膜肽-家蝇天蚕素(HTPP-MDC)融合多肽体外对乙型肝炎病毒(HBV)的抑制作用及其在肝细胞中的定位。方法:不同浓度HTPP-MDC分别与HepG2.2.15细胞和Chang liver细胞共培养,用四甲基偶氮唑盐(MTT)比色法检测药物对细胞的毒性作用,应用ELISA和实时荧光定量PCR技术定量研究HTPP-MDC对HepG2.2.15细胞系分泌HBsAg、HBeAg及HBV DNA复制的影响。应用激光共聚焦技术研究HTPP-MDC在肝细胞中的定位。结果:HTPP-MDC在体外能有效抑制HepG2.2.15细胞系分泌HBsAg和HBeAg,对HBVDNA的复制亦有显著抑制作用。激光共聚焦显微镜观察显示HTPP-MDC进入细胞内部。结论:HTPP-MDC在体外对HBV复制有较强的抑制作用,并能快速透过细胞膜进入细胞内,有望用于临床治疗慢性乙型肝炎相关疾病。AIM： To investigate the antiviral effect of hepatocyte-targeting and cell-penetrating peptide and Musca domestica cecropin（HTPP-MDC） fusion polypeptide against hepatitis B virus（HBV） in vitro,and to observe the penetrating ability of HTPP-MDC in hepatocytes.METHODS： HepG2.2.15 cells and Chang liver cells were co-cultured in vitro with HTPP-MDC at different concentrations.MTT assay was used to detect the cytotoxicity of HTPP-MDC in vitro.The levels of HBsAg,HBeAg and HBV DNA in HepG2.2.15 cell culture supernatants were measured by ELISA and realtime fluorescence quantitative PCR.The penetrating ability of HTPP-MDC was detected by laser confocal microscopy.RESULTS： The levels of HBsAg,HBeAg and HBV DNA in the supernatants of HepG2.2.15 cells treated with HTPP-MDC were remarkably reduced.The inhibitory effect of HTPP-MDC depended on the dose and action time of the drug.FITC-labeled HTPP-MDC was observed inside the cells under laser confocal microscope.CONCLUSION： HTPP-MDC strongly inhibits HBV replication in HepG2.2.15 cells.The penetrating ability of HTPP-MDC into hepatocytes indicates that HTPPMDC is useful in clinic therapy for chronic hepatitis B-related diseases in the future.

关 键 词：肝细胞靶向穿膜肽 家蝇天蚕素 融合多肽 肝炎病毒 乙型 

分 类 号：R965[医药卫生—药理学]