灰树花多糖抑制乙型肝炎病毒活性的研究

Study on Inhibit Activity by Maitake Polysaccharide to HBV Virus

用水提取法从新鲜灰树花子实体中提取灰树花多糖(Maitake Polysaccharide,MP),分别以不同质量浓度MP作用于HepG2.2.15细胞,MTT法测定细胞生长抑制率,计算半数细胞毒性浓度(CC50)。另以不同质量浓度的MP和0.1 mg/m L拉米夫定处理HepG2.2.15细胞,采用酶联免疫法(ELISA法)检测分别处理3、6、9 d的细胞培养上清液中乙肝表面抗原(HBsAg)和e抗原(HBeAg)滴度,用荧光定量PCR法检测处理9 d后的细胞培养上清液中HBV DNA拷贝数,分别计算对HBsAg、HBeAg和HBV DNA的抑制率和半数抑制质量浓度(IC_(50))及治疗指数(TI50)。结果表明,MP对HepG2.2.15细胞的CC50为5.154 mg/m L。对HBsAg、HBeAg及HBV DNA的抑制率呈剂量依赖性,给药9 d后的IC_(50)分别是4.814、0.806、0.621 mg/m L。TI50分别为HBsAg1.071,HBeAg 6.398和HBV DNA 8.306。因此MP具有低毒特点并对体外培养的HBV复制和表达具有较明显的抑制作用。

To investigate influence ofMaitake Polysaccharide（MP） on expression and replication of HBV in HepG2.2.15 cell. MP from fresh Maitake fruiting bodies was extracted by water. The cell growth inhibition rate of HepG2.2.15 cells was detected by MTT assay in 4,2,1,0.5,0.25 mg/mL concentration respectively and the half cell toxicity concentration （CC50） was calculated. HepG2.2.15 cells were treated by MP with different concentration or 0.1 mg/mL lamivudine, and the Hepatitis B surface antigen and e antigen （HBsAg and HBeAg） in cell supematant were evaluated byenzyme-linked immunoassay assay（ELISA）, HBV DNA contents after cells were treated 9 days were detected using quantitative PCR method. Then each inhibition rate, the half inhibition concentration （IC50） and the therapeutic index （TI50） were calculated respectively based on the data. The CC50 of MP treated to HepG2.2.15 cells is 5.154 mg/mL. The inhibition rate of MP in HBsAg, HBeAg and of HBV DNA are dose dependent and the IC50 are 4.814,0.806,0.621 mg/mL,TI50 are 1.071,6.398 and 8.306 respectively. MP has strong inhibitory effect on HBV replication and expression in vitro.