玉郎伞多糖抑制HepG 2.2.15细胞HBV的复制和表达的研究

Inhibition of YuLangSan Polysaccharide on the Replication and Expression of HepG 2.2.15 Cells HBV

目的研究玉郎伞多糖(YuLangSan Polysaccharide)的体外抗乙型肝炎病毒(HBV)的作用。方法采用HepG 2.2.15细胞模型进行体外培养,给予不同浓度玉郎伞多糖,以拉米夫定(Lamivudine)作阳性药对照,作用72 h, 144 h后,采用实时荧光定量PCR(Quantitative Real-time, PCR)检测HBV DNA的水平,采用酶联免疫吸附试验(enzyme-linked immunosorbent assay, ELISA)检测HBsAg、HBeAg的滴度,观察玉郎伞多糖对HepG 2.2.15细胞HBV DNA的复制及病毒抗原表达的影响,同时以MTT法检测玉郎伞多糖在体外对HepG 2.2.15细胞的生长抑制作用,评价玉郎伞多糖抗HBV作用。结果玉郎伞多糖对HepG 2.2.15细胞的半数毒性浓度(TC50)为113.28 mg·L^(-1);可以显著抑制HBV DNA的复制;对HepG 2.2.15细胞分泌HBsAg的半数抑制浓度(IC50)为3.87 mg·L^(-1),其治疗指数(TI)为29.27;对HBeAg的IC50为7.53 mg·L^(-1),TI为15.05;玉郎伞多糖在0.625,1.25,2.5,5.0,10 mg·L^(-1)浓度作用HepG 2.2.15细胞144 h后能显著抑制HBsAg和HBeAg的表达,其最高抑制率分别为61.54%,59.46%。结论玉郎伞多糖在体外有较强的抗HBV的作用,且安全性较高。

OBJECTIVE To observe the effect of polysaccharide from YuLangSan on anti-hepatits B virus in vitro. METHODS HepG 2.2.15 cells wer cultured in vitro and treated by different concentrations YuLangSan polysaccharide.Lamivudine was used as positive control.After treated by YuLangSan polysaccharide, the HBV DNA level in hepatic tissues was quantified by PCR method, and the titer of HBsAg and HBeAg were evaluated by enzyme-linked immunosorbent assay(ELISA) to observed the influence of YuLangSan polysaccharide on HBV DNA replication and viral antigen expression in HepG 2.2.15.The inhibition of YuLangSan polysaccharide on HepG 2.2.15 cells were detected by means of MTT to evaluate the anti-HBV effect. RESULTS The half toxic concentration(TC50) of YuLangSan polysaccharide on HepG 2.2.15 cells was 113.28 mg·L^(-1).YuLangSan polysaccharide suppressed the levels of HBV DNA replication effectively.The half inhibitory concentration(IC50) of YuLangSan polysaccharide on HBsAg was 3.87 mg·L^(-1).The treatment index(TI) of YuLangSan polysaccharide on HBsAg was 29.27.And the IC50 of YuLangSan polysaccharide on HBeAg was 7.53 mg·L^(-1),TI was 15.05.The results indicated that YuLangSan Polysaccharide suppressed HBsAg and HBeAg expressions significantly with the maximum inhibition ratios of 61.54% and 59.46% after treated with 0.625,1.25,2.5,5.0 and 10 mg·L^(-1) for 144 hours. CONCLUSION YuLangSan polysaccharide can significantly inhibit HBV in vitro,and the safe is high.