锌、铁饱和乳铁蛋白体外抗HBsAg分泌的研究

Study of inhibition effect on hepatitis B surface antigen of zinc-, iron-saturated bovine lactoferrin in vitro

目的:探讨锌饱和乳铁蛋白(Zn2+-BLF)、铁饱和乳铁蛋白(Fe2+-BLF)体外抑制乙型肝炎表面抗原分泌作用,为锌、铁乳铁蛋白应用于临床治疗乙型肝炎病毒(HBV)感染提供理论和试验依据。方法:以天然HBV感染HepG2细胞为模型,通过应用ELISA法测定细胞上清中的HBsAg水平来检测Zn2+-BLF、Fe2+-BLF的抗HBsAg分泌效果,并用MTT法对Zn2+-BLF、Fe2+-BLF对HepG2细胞的毒性进行研究。结果:Zn2+-BLF、Fe2+-BLF对细胞的最大无毒剂量(TD0)分别为1.5g/L、3.0g/L;先用HBV对HepG2细胞进行感染,再分别加入Zn2+-BLF、Fe2+-BLF,各浓度组Zn2+-BLF均对HBsAg分泌有一定抑制作用,Zn2+-BLF浓度1.0g/L时对HBsAg的抑制率达60.49%;浓度为1.0g/L、0.5g/L的Fe2+-BLF能显著抑制HBsAg的分泌,但0.1g/L的Fe2+-BLF不能显著抑制HBsAg的分泌。结论:当HepG2细胞感染HBV后,Zn2+-BLF、Fe2+-BLF可以显著抑制HBsAg的分泌,Zn2+-BLF对HBsAg的抑制作用强于Fe2+-BLF,但其作用机理有待深入研究。

Objective： To probe inhibition effect on hepatitis B surface antigen （HBsAg） of zinc-saturated bovine lactoferrin （Zn^2＋-BLF） and iron-saturated bovine lactoferrin （Fe^2＋ -BLF） in vitro, so as to provide theoretical as well as experimental basis for the clinical management of Zn^2＋-BLF, Fe^2＋-BLF infection. Methods： Nature hepatitis B virus（HBV） was used to infect HepG2 cell as model, HBsAg in the culture supernatant was measured by ELISA to detect the effect of Zn^2＋-BLF, Fe^2＋-BLF on HBsAg secretion, and Ml-r test was used to examine cytotoxic effect of Zn^2＋ -BLF, Fe^2＋ -BLF. Result： The maximum nontoxic dose （TDo） in HepG2 cell of Zn^2＋ -BLF and Fe^2＋ -BLF are respective 1.5g/L, 3.0g/L; After HepG2 cell were infected with HBV, every Zn^2＋-BLF test group had effect on inhibiting HBsAg to some extent. At the highest concentration（1.0g/L） BLF was most effective, exhibiting 60.49% inhibition; concentration of 1.0g/L, 0.5g/L Fe^2＋ -BLF could significantly inhibit HBsAg, but concentration of 0.1g/L test group couldn＇t. Conclusion： After HepG2 cell was infected with HBV, HBsAg could be inhibited by Zn^2＋ -BLF and Fe^2＋ -BLF, Zn^2＋ -BLF has a higher inhibition effect than Fe^2＋ -BLF, but the mechanism need to be lucubrated.