纤维结合素对HBV感染原代培养人胎肝细胞的影响  

Effect of fibronectin on infection of primary human fetal hepatocytes with HBV in vitro

作　　者：王方[1] 王宇明[1] 王小红[1] 汤勃[1] 刘国栋[1] 刘俊[1] 

机构地区：[1]第三军医大学西南医院,重庆400038

出　　处：《解放军医学杂志》2004年第5期400-402,共3页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金资助课题 (编号 30 1 70 847)

摘　　要：目的　研究纤维结合素对HBV体外感染原代培养人胎肝细胞的影响。方法　用纤维结合素包被培养皿 ,利用从HBV阳性血清纯化的HBV病毒颗粒感染体外培养的人胎肝细胞。应用ELISA法检测培养上清中HBsAg和HBeAg,免疫组化法检测细胞核中HBcAg,荧光定量PCR法检测细胞中HBVDNA ,巢式PCR法检测细胞中cccDNA ,并和未包被的胎肝细胞进行对照。结果　未包被时上清中HBsAg和HBeAg于第 5日出现阳性 ,包被后自第 1日起出现阳性 ;未包被时胎肝细胞核中HBcAg于感染后第 2日起出现阳性 ,阳性率为 10 %～ 15 % ,包被后自第 1日起出现阳性 ,阳性率达 90 %以上 ;未包被时细胞中HBVDNA定量自第 4日起检出 ,包被后自第 1日起检出 ;未包被时细胞中HBVcccDNA自第 8日出现阳性 ,包被后自第 2日起出现阳性。结论　纤维结合素包被可以促进HBV体外感染原代培养人胎肝细胞 ,使HBV感染胎肝细胞的时间提前 。Objective This experiment was carried out to study the effect of fibronectin on the infection of primary human fetal hepatocytes with hepatitis B virus (HBV) in vitro. Method Human fetal hepatocytes were inoculated with HBV positive serum in culture dish coated with fibronectin (Fn) in vitro. HBsAg and HBeAg in supernatant were assayed with ELISA, HBcAg in nuclei by immunohistochemistry, HBV DNA in cells by fluorescence quantitative PCR, and cccDNA in cultured cells by nest PCR. The results were compared with the cells cultured in dishes without fibronectin coating. Results HBsAg and HBeAg in supernatants were positive on the fifth day in dishes without Fn coating, and on the first day in dishes with Fn coating. HBcAg in nuclei was positive on the second and the first day in dishes without and with Fn coating, respectively, and the positive rates were about 10%～15% and 90%, respectively. HBV DNA was detected in cells on the fourth day and first day in dishes without and with Fn coating, respectively. CccDNA in cells was positive on the eighth day and the first day in dishes without and with Fn coating, respectively. Conclusion Fibronectin coating could facilitate HBV infection in primary fetal hepatocytes in vitro. The infection time of HBV was shortened, and the number of infected cells was increased.

关 键 词：纤维结合素 肝炎病毒 乙型 人胎肝细胞 细胞培养 

分 类 号：R322.47[医药卫生—人体解剖和组织胚胎学]