灵猫方通过增强天然免疫功能抑制乙型肝炎病毒复制的研究  

The Lingmao Formula Suppresses the Replication of Hepatitis B Virus by Enhancing Innate Immune Response

作　　者：朱晓骏[1] 张鑫[2] 周振华[2] 金树根[2] 高雅婷 孙学华[1] 杨婉凤[1] 高月求[1] 

机构地区：[1]上海中医药大学附属曙光医院肝病科,上海201203 [2]上海中医药大学附属曙光医院细胞免疫实验室

出　　处：《中西医结合肝病杂志》2018年第1期33-36,共4页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：国家自然科学基金(No.81403351;81774256;81503545;81673935;81473629);上海市自然基金(No.14ZR1441900);上海市中医药事业发展三年行动计划项目(No.ZY3-CCCX-3-3029);上海市科委扬帆计划(No.15YF1412300;14YF1411600)

摘　　要：目的:研究灵猫方对pHBV1.3转染Hep G2细胞的天然免疫功能的影响,探讨灵猫方抗乙肝病毒(HBV)的作用机制。方法:制备0.25mg/ml和0.5mg/ml灵猫方水提物干预HBV质粒转染HepG2细胞,以0.9%Na Cl溶液作为空白对照组,72h后收集培养上清液,ELISA法检测HBsAg和HBeAg水平;收集细胞提取总RNA,RT-PCR法检测细胞内IFN-α、IFN-β、IFN-λ1及干扰素效应蛋白OAS1、Mx A、PRKmRNA表达水平,RIG-I mRNA表达水平。结果:与空白对照组比较,0.5mg/ml灵猫方干预pHBV1.3转染Hep G2细胞分泌的HBsAg和HBeAg水平明显降低,细胞内IFN-α、IFN-β、IFN-λ1 mRNA及OAS1、Mx A、PRKmRNA表达水平表明显升高(P<0.01),RIG-I mRNA表达水平明显升高(P<0.01)。结论:灵猫方通过促进pHBV1.3质粒转染HepG2细胞的I、III型干扰素产生,增强天然免疫功能,从而抑制HBV复制,可能是通过RIG-I通路诱导干扰素产生,增加干扰素的形成。Objective：To observe the effect of Lingmao Formula on innate immune response in HepG2 cells transfected with plasmid pH- BV1.3, so as to illuminate the mechanism of the anti-HBV. Methods： The aqueous extract of Lingmao Formula was prepared. 0. 25 mg,/ml and 0. 5 mg/ml Lingmao Formula were intervened HepG2 celltransfected with plasmid pHBVl. 3 cells andO. 9% NaC1 solution was used as control group, respectively for 72 hours. The levels of HBsAg and HBeAg in supernatant were detected by ELISA. The mRNA levels of IFN- α, IFN-β, IFN-λ1, OAS1, MxA, PRK and RIG-I in the cells were detected by RT-PCR. Results： Compared with the control group, the HBsAg and HBeAg levels of Lingmao Formula group in HepG2 cell transfected with plasmid pHBV1.3 cells were significantly decreased, the expression levels of intracellular IFN-α, IFN-β, IFN-λ1, OAS1, MxA, PRK and RIG-I mRNA were significantly increased （P 〈 0.01 ）. Conclusion： Lingmao Formula may inhibit HBV replication by enhancing innate immune response because of RIG-I interferon - induced pathway in HepG2 cells transfected with pHBV1.3.

关 键 词：灵猫方 HEPG2 天然免疫功能 RIG-I 

分 类 号：R285[医药卫生—中药学]