基于TLR3补肾健脾方水提物抑制乙肝病毒复制机制研究

朱晓骏; 郑超; 张鑫; 郑彦希; 余卓; 孙学华; 高月求

doi:10.16306/j.1008-861x.2022.S1.035

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基于TLR3补肾健脾方水提物抑制乙肝病毒复制机制研究

实验研究 | 更新时间：2022-08-29

- 基于TLR3补肾健脾方水提物抑制乙肝病毒复制机制研究
- Mechanism study on inhibition of hepatitis B virus replication by TLR3 based water extract of Bushen Jianpi Decoction
- 上海中医药大学学报 2022年36卷第S1期页码：149-153
- 作者机构：
  
  1.上海中医药大学附属曙光医院肝病科（上海　201203）
- 作者简介：
  
  朱晓骏，女，博士，副主任医师，主要从事中西医结合治疗慢性乙型肝炎临床与基础研究
  高月求，主任医师，教授；E-mail：gaoyueqiu0418@163.com
- 基金信息：
  
  国家自然科学基金资助项目(81673935;81774240;81774256;81904164;81973773);国家“艾滋病和病毒性肝炎等重大传染病防治”科技重大专项(2018ZX10725504);浦东新区中医联合体建设项目(PDZY-2019-0601);上海市中医药事业发展三年行动计划（ZY〔2018-2020〕-CCCX-2003-01）;重大疑难疾病中西医临床协作试点项目（ZY〔2018-2020〕-FXTW-2004）
- DOI：10.16306/j.1008-861x.2022.S1.035
  中图分类号：
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朱晓骏,郑超,张鑫等.基于TLR3补肾健脾方水提物抑制乙肝病毒复制机制研究[J].上海中医药大学学报,2022,36(S1):149-153.

ZHU Xiaojun,ZHENG Chao,ZHANG Xin,et al.Mechanism study on inhibition of hepatitis B virus replication by TLR3 based water extract of Bushen Jianpi Decoction[J].Academic Journal of Shanghai University of Traditional Chinese Medicine,2022,36(S1):149-153.
朱晓骏,郑超,张鑫等.基于TLR3补肾健脾方水提物抑制乙肝病毒复制机制研究[J].上海中医药大学学报,2022,36(S1):149-153. DOI： 10.16306/j.1008-861x.2022.S1.035.

ZHU Xiaojun,ZHENG Chao,ZHANG Xin,et al.Mechanism study on inhibition of hepatitis B virus replication by TLR3 based water extract of Bushen Jianpi Decoction[J].Academic Journal of Shanghai University of Traditional Chinese Medicine,2022,36(S1):149-153. DOI： 10.16306/j.1008-861x.2022.S1.035.

摘要

目的,2,通过 pHBV1.3-pCR2.1质粒转染HepG2细胞探讨补肾健脾方水提物抑制乙肝病毒的免疫机制。,方法,2,250 μg/ml、500 μg/ml补肾健脾方水提物干预 pHBV1.3-pCR2.1质粒转染HepG2细胞，ELISA法检测细胞上清HBsAg、HBeAg含量；500 μg/ml补肾健脾方水提物干预pHBV1.3-pCR2.1质粒转染HepG2细胞，Westen blot法检测Toll样受体3（TLR3）、p-IRF3蛋白表达，qRT-PCR法检测细胞内,IFN,-,β, mRNA表达；补肾健脾方干预HepG2细胞，pHBV1.3-pCR2.1质粒转染HepG2细胞，TLR13 siRNA下调TLR3表达，Westen blot法检测TLR3、p-IRF3蛋白表达，qRT-PCR法检测细胞内,IFN,-,β, mRNA 表达。,结果,2,500 μg/ml补肾健脾方与空白对照组、250 μg/ml补肾健脾方比较，HBsAg均明显下降（,P<,0.05，,P<,0.01）；500 μg/ml补肾健脾方与空白对照组、250 μg/ml补肾健脾方比较，HBeAg均明显下降（,P<,0.05，,P<,0.01）；补肾健脾方（500 μg/ml）可以增强TLR3、p-IRF3蛋白表达，促进细胞内,IFN,-,β, mRNA 表达（,P,<,0.01）；补肾健脾方干预 pHBV1.3-pCR2.1质粒转染HepG2细胞，TLR3 siRNA抑制细胞TLR3蛋白表达，p-IRF3蛋白表达明显下降，同时细胞内,IFN,-,β, mRNA 表达也明显下降（,P<,0.01）。,结论,2,补肾健脾方水提物通过增强 pHBV1.3-pCR2.1质粒转染HepG2细胞TLR3作用，促进细胞内IFN-β分泌，从而抑制乙型肝炎病毒的复制。

Abstract

Objective： To explore the immune mechanism on the water extract of Bushen Jianpi Decoction in inhibiting hepatitis B virus in HepG2 cells transfected with plasmid pHBV1.3-pCR2.1.,Methods,2,After the water extract of Bushen Jianpi Decoction intervened in HepG2 cells transfected with plasmid pHBV1.3-pCR2.1 in two different concentrations（250 μg/ml and 500 μg/ml）， HBsAg and HBeAg contents in cell supernatant were detected by ELISA. After the water extract of Bushen Jianpi Decoction intervened in HepG2 cells transfected with plasmid pHBV1.3-pCR2.1 in concentrations of 500 μg/ml， the expressions of TRL3， p-IRF3 protein were detected by WB， and the expression of ,IFN,-,β, mRNA was detected by qRT-PCR. Bushen Jianpi Decoction intervened in HepG2 cells led that HepG2 cells were transfected with plasmid pHBV1.3-pCR2.1 and TLR3 siRNA down regulated the expression of TLR3. The expressions of TLR3 and p-IRF3 protein were detected by Western blot， and the expression of ,IFN,-,β, mRNA was detected by qRT-PCR.,Results,2,HBsAg of Bushen Jianpi Decoction（500 μg/ml）was significantly decreased compared with that in Bushen Jianpi Decoction（250 μg/ml）and the model group（,P<,0.05，,P<,0.01）. HBeAg of Bushen Jianpi Decoction（500 μg/ml）was significantly decreased compared with that in Bushen Jianpi Decoction（250 μg/ml）and the model group（,P<,0.05，,P<,0.01）. Bushen Jianpi Decoction（500 μg/ml） enhanced TLR3 and p-IRF3 protein expressions， and promoted ,IFN⁃β, mRNA expression in cell（,P<,0.01）. After Bushen Jianpi Decoction intervened in HepG2 cells transfected with plasmid pHBV1.3-pCR2.1， TLR3 siRNA inhibited TLR3 protein expression and p-IRF3 protein expression was decreased significantly and the expression of intracellular ,IFN⁃β ,mRNA expression was decreased significantly.,Conclusion,2,The water extract of Bushen Jianpi Decoction can inhibit hepatitis B virus replication by enhancement of TLR3 effect in HepG2 cells transfected with plasmid pHBV1.3-pCR2.1 and promotion of intracellular IFN-β secretion.

关键词

补肾健脾方水提物乙型肝炎病毒Toll样受体3

Keywords

water extract of Bushen Jianpi Recipehepatitis B virusToll-like receptor 3

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