恩替卡韦治疗后，肝细胞内质网应激抑制乙型肝炎病毒的分

StephenW

Hepatocyte Endoplasmic Reticulum Stress Inhibits Hepatitis B Virus Secretion and Delays Intracellular Hepatitis B Virus Clearance After Entecavir Treatment
Huan Chen  1 , Maoyuan Mu  1 , Qichuan Liu  1 , Han Hu  1 , Caiyun Tian  1 , Guoyuan Zhang  1 , Ying Li  1 , Fangwan Yang  1 , Shide Lin  1
Affiliations
Affiliation

    1
    Department of Infectious Diseases, Affiliated Hospital of Zunyi Medical University, Zunyi, China.

    PMID: 33614671 PMCID: PMC7890007 DOI: 10.3389/fmed.2020.589040

Abstract

Background: The aim of this study was to explore the effects of endoplasmic reticulum (ER) stress on hepatitis B virus (HBV) replication and the antiviral effect of entecavir (ETV). Methods: Thapsigargin (TG) and stearic acid (SA) were used to induce ER stress in HepG2.2.15 cells and HepAD38 cells that contained an integrated HBV genome, while ETV was used to inhibit HBV replication. The expression levels of glucose-regulated protein 78 (GRP78) and phosphorylated eukaryotic translation initiation factor 2 subunit alpha (p-eIF2α) were measured by western blotting. Intracellular HBV DNA was determined by qPCR; HBsAg by western blotting; HBV RNA by real-time RT-qPCR; HBsAg and HBeAg in supernatants by enzyme-linked immunosorbent assay (ELISA); and HBV DNA in supernatants by qPCR. Results: TG and SA induced ER stress in HepG2.2.15 cells and HepAD38 cells from 12 to 48 h post treatment. However, 4-phenylbutyric acid (PBA) partly alleviated the TG-induced ER stress. Moreover, TG inhibited HBsAg, HBeAg, and HBV DNA secretion from 12 to 48 h, while different concentrations of SA inhibited HBsAg and HBV DNA secretion at 48 h. TG promoted intracellular HBV DNA and HBsAg accumulation and the transcription of the HBV 3.5-kb mRNA and S mRNA. PBA treatment restored the secretion of HBsAg and HBV DNA. Finally, ER stress accelerated extracellular HBV DNA clearance but delayed intracellular HBV DNA clearance after ETV treatment. Conclusions: Hepatocyte ER stress promoted intracellular HBV DNA and HBsAg accumulation by inhibiting their secretion. Our study also suggested that hepatocyte ER stress delayed intracellular HBV DNA clearance after ETV treatment.

Keywords: HBV DNA; HBsAg; HepG2215; endoplasmic reticulum stress; stearic acid; thapsigargin.

Copyright © 2021 Chen, Mu, Liu, Hu, Tian, Zhang, Li, Yang and Lin.

StephenW

恩替卡韦治疗后，肝细胞内质网应激抑制乙型肝炎病毒的分泌并延迟细胞内乙型肝炎病毒的清除
陈欢1，茂源木1，刘启川1，胡虎1，田彩云1，张国元1，李颖1，杨芳婉1，林实德1
隶属关系
联系

    1个
    遵义医科大学附属医院传染病科，四川遵义。

    PMID：33614671 PMCID：PMC7890007 DOI：10.3389 / fmed.2020.589040

抽象的

背景：这项研究的目的是探讨内质网（ER）应激对乙型肝炎病毒（HBV）复制和恩替卡韦（ETV）的抗病毒作用。方法：用毒胡萝卜素（TG）和硬脂酸（SA）在含有完整HBV基因组的HepG2.2.15细胞和HepAD38细胞中诱导内质网应激，而ETV抑制HBV复制。通过蛋白质印迹法测量葡萄糖调节蛋白78（GRP78）和磷酸化的真核翻译起始因子2亚基α（p-eIF2α）的表达水平。通过qPCR确定细胞内HBV DNA；免疫印迹法检测HBsAg；实时RT-qPCR检测HBV RNA；酶联免疫吸附法（ELISA）测定上清液中的HBsAg和HBeAg； qPCR检测上清液中的HBV DNA。结果：TG和SA在治疗后12至48小时诱导HepG2.2.15细胞和HepAD38细胞的内质网应激。但是，4-苯基丁酸（PBA）部分缓解了TG诱导的ER应激。此外，TG在12至48小时内抑制HBsAg，HBeAg和HBV DNA分泌，而不同浓度的SA在48 h抑制HBsAg和HBV DNA分泌。 TG促进细胞内HBV DNA和HBsAg的积累以及HBV 3.5-kb mRNA和S mRNA的转录。 PBA治疗恢复了HBsAg和HBV DNA的分泌。最后，内质网应激可促进ETV治疗后细胞外HBV DNA清除，但延迟细胞内HBV DNA清除。结论：肝细胞内质网应激可通过抑制其分泌促进细胞内HBV DNA和HBsAg的积累。我们的研究还表明，ETV治疗后，肝细胞内质网应激会延迟细胞内HBV DNA清除。

关键字：HBV DNA；乙肝表面抗原HepG2215；内质网应激硬脂酸;毒胡萝卜素。

版权所有©2021 Chen，Mu，Liu，Hu，Tian，Zhang，Li，Yang and Lin。

StephenW

https://www.frontiersin.org/arti ... ed.2020.589040/full