锌、铁、锰饱和乳铁蛋白体外抑制乙型肝炎病毒DNA的研究

Study of inhibition effect of zinc-,iron-and manganese-saturated bovine lactoferrin on hepatitis B virus DNA in vitro

目的探讨锌、铁、锰饱和乳铁蛋白体外抑制乙型肝炎病毒DNA(HBV-DNA)作用。方法以天然HBV感染HepG2细胞为模型,通过应用荧光定量PCR法测定细胞中HBV-DNA水平来检测锌、铁、锰饱和乳铁蛋白抑制HBV效果,并应用四甲基偶氮唑盐比色法(MTT)进行锌、铁、锰饱和乳铁蛋白对HepG2细胞的毒性研究。结果锌、铁、锰饱和乳铁蛋白对细胞的最大无毒剂量(TD0)分别为1.5、3.0和1.5g/L;用HBV感染HepG2细胞,分别加入锌、铁、锰饱和乳铁蛋白,各实验组对HBV-DNA均有显著抑制作用,浓度为1.5g/L的锌、铁、锰饱和乳铁蛋白抗HBV-DNA结果经对数转换后分别为:(5.746±0.114)、(6.446±0.103)和(5.999±0.725)。结论当HepG2细胞感染乙肝病毒后,锌、铁、锰饱和乳铁蛋白可以显著抑制HBV-DNA,抑制作用强弱依次为:锌饱和乳铁蛋白>锰饱和乳铁蛋白>铁饱和乳铁蛋白。

Objective To probe inhibition effect of zinc-saturated（Zn 2＋-BLF）,iron-saturated（Fe 2＋-BLF） and manganese-saturated（Mn 2＋-BLF） bovine lactoferrin on hepatitis B virus DNA（HBV-DNA） in vitro.Methods Nature HBV was used to infect HepG2 cell,HBV-DNA was measured by fluorescence polymerase chain reaction to detect the effect of Zn 2＋-BLF,Fe 2＋-BLF and Mn 2＋-BLF,the MTT test was used to examine cytotoxic effect of Zn 2＋-BLF,Fe 2＋-BLF and Mn 2＋-BLF.Results The maximum nontoxic dose（TD_0） in HepG2 cell of Zn 2＋-BLF,Fe 2＋-BLF and Mn 2＋-BLF were 1.5,3.0 and 1.5g/L respective.After HepG2 cell were infected with HBV,every Zn 2＋-BLF,Fe 2＋-BLF and Mn 2＋-BLF test group had effect on inhibiting HBV-DNA to some extent.Compared the inhibition effect of concentration of 1.5g/L test group in each term,the inhibition result of Zn 2＋-BLF was 5.746±0.114,the inhibition result of Fe 2＋-BLF was 6.446±0.103,and the inhibition result of Mn 2＋-BLF was 5.999±0.725.Conclusion After HepG2 cell was infected with HBV,HBV-DNA could be inhibited by Zn 2＋-BLF,Fe 2＋-BLF and Mn 2＋-BLF.Zn 2＋-BLF has higher inhibition effect than Mn 2＋-BLF,and Fe 2＋-BLF has the lowest inhibition effect,but the possible mechanism need to be lucubrated.