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目的比较天然提取与重组高迁移率组蛋白N2(HMGN2)的抗乙型病毒性肝炎病毒(乙肝病毒,HBV)活性,以探讨HMGN2分子的抗HBV作用。方法以HBV DNA转染的细胞株Hep G 2.2.15细胞为体外抗HBV模型,用不同浓度的天然提取和重组HMGN2分子作用于Hep G 2.2.15细胞,分别在第4,8天收集细胞培养上清液,酶联免疫吸附(ELISA)法检测上清中HBV表面抗原(HBs Ag)及e抗原(HBe Ag),用实时荧光定量聚合酶链式反应(PCR)法检测上清液HBV DNA的含量。结果天然提取HMGN2与重组HMGN2在1~100mg·L-1对Hep G 2.2.15细胞无细胞毒性;天然提取HMGN2与重组HMGN2蛋白分子在1~5 mg·L-1水平即可显著抑制HBe Ag和HBs Ag的表达,可显著降低HBV DNA拷贝数,继续增加剂量,抗病毒效果不再明显增加,且在相同剂量下比较,两者作用差异无统计学意义。结论天然提取HMGN2与重组HMGN2分子在体外细胞培养中具有同样直接抗HBV活性。
Objective To compare the anti-hepatitis B virus (HBV) activity of natural extract and recombinant high-mobility group histone N2 (HMGN2) in order to explore the anti-HBV effect of HMGN2 molecule. Methods HepG 2.2.15 cells transfected with HBV DNA were treated with anti-HBV in vitro. HepG 2.2.15 cells were treated with different concentrations of natural extract and recombinant HMGN2. Cells were harvested on day 4 and day 8 respectively The supernatant and enzyme-linked immunosorbent assay (ELISA) were used to detect HBs Ag and HBe Ag in the supernatant. The supernatant HBV DNA was detected by real-time fluorescence quantitative polymerase chain reaction (PCR) Content. Results The natural extraction of HMGN2 and recombinant HMGN2 at 1 ~ 100 mg · L-1 had no cytotoxicity on Hep G 2.2.15 cells. The natural extraction of HMGN2 and recombinant HMGN2 at 1 ~ 5 mg · L-1 could significantly inhibit the growth of HBe Ag And HBs Ag expression could significantly reduce HBV DNA copy number, continue to increase the dose, anti-virus effect is no longer significantly increased, and in the same dose comparison, the role of the difference was not statistically significant. Conclusion The natural extract of HMGN2 and recombinant HMGN2 molecules have the same direct anti-HBV activity in cell culture in vitro.