摘要
目的:探讨β-LPA体外抗乙型肝炎病毒(hepatitis B virus,HBV)的作用.方法:通过β-LPA干预培养HepG2.2.2.15细胞,ELISA法检测上清HBsAg和HBeAg,P标32记HBV DNA为探针,Southern blot法检测细胞内的HBV DNA,再以计算机图像处理进行定量分析,得出50%抑制的药物浓度(ED50),以MTT法检测不同浓度药物的细胞毒性,求出50%细胞抑制的药物浓度(ID50).结果:β-LPA体外明显抑制HBV DNA的复制,并呈浓度依赖性.ED50为0.01μmol/L,β-LPA细胞毒性实验显示ID50为50μmol/L.低浓度的β-LPA对上清HBsAg,HBeAg无明显影响,高浓度时有显著的抑制作用.结论:β-LPA具有明显的体外抑制病毒DNA复制作用,且细胞毒性小.
AIM： To explore the in vitro effects of a novel nucleoside analog （β-LPA） against hepatitis B virus （HBV）.
METHODS： HepG2.2.2.15 cells were cultured and treated with various concentrations of β-LPA. Serum HBsAg and HBeAg were determined by enzyme-linked imrnunosorbent assay （ELISA）. Intracelluar DNA was extracted and subjected to Southern blotting, hybridized with 32p-labeled HBV probe and autoradiographed. The intensity of the autoradiographic bands was quantified by densitometric scans of computer and ED50 value was calculated. Cytotoxicity with different concentrations of drugs was examined by methyl thiazolyl tetrazolium （MTT） method and ID50 value was calculated. RESULTS： Autoradiographic bands showed that β-LPA inhibited the replication of HBV DNA in a dose-dependent manner. ED50 value was 0.01μmol/L. Cytotoxicity experiment showed that the ID50 value of β-LPA was 50μmol/L. The contents of HBsAg and HBeAg were decreased in a concentration-dependent manner.β-LPA at a low concentration had no marked effect on HBsAg and HBeAg in supernate. CONCLUSION： β-LPA possesses potent inhibitory effect on the replication of HBV in vitro with little cytotoxicity.
出处
《世界华人消化杂志》
CAS
北大核心
2008年第10期1110-1113,共4页
World Chinese Journal of Digestology
基金
国家自然科学基金资助项目
No.30330680~~
