Anti-HBV Effect and Mechanism of Sophora Alkaloids Combined with Entecavir

CHEN Jia-xin; SHEN Hong-hui; LIU Xiao-qiong; WANG Jia-bo; ZOU Wen-jun; WANG Shi-yu; XIAO Xiao-he

doi:10.13422/j.cnki.syfjx.2017010107

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Anti-HBV Effect and Mechanism of Sophora Alkaloids Combined with Entecavir

更新时间：2024-01-04

- Anti-HBV Effect and Mechanism of Sophora Alkaloids Combined with Entecavir
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 23, Issue 1, Pages: 107-113(2017)
- 作者机构：
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- DOI：10.13422/j.cnki.syfjx.2017010107
  CLC： R96
- Published：2017
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CHEN Jia-xin, SHEN Hong-hui, LIU Xiao-qiong, et al. Anti-HBV Effect and Mechanism of Sophora Alkaloids Combined with Entecavir[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(1): 107-113.
DOI：

CHEN Jia-xin, SHEN Hong-hui, LIU Xiao-qiong, et al. Anti-HBV Effect and Mechanism of Sophora Alkaloids Combined with Entecavir[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(1): 107-113. DOI： 10.13422/j.cnki.syfjx.2017010107.

摘要

目的：研究苦参碱类生物碱联合恩替卡韦抗耐药乙型肝炎病毒（HBV）的作用效果以及对p38 MAPK，钠离子-牛磺胆酸共转运蛋白（NTCP）表达的影响。方法：以解放军第302医院自行构建的恩替卡韦耐药乙型肝炎病毒复制细胞株HepG2.A64为模型，采用CPE，CCK-8分别考察苦参碱类生物碱氧化苦参碱、苦参碱、槐果碱、槐定碱和核苷类药物恩替卡韦单独用药及两者联合用药的细胞毒性；采用酶联免疫法（ELISA），实时荧光定量聚合酶链式反应（Real-time PCR），蛋白质免疫印迹法（Western blot）分别观察联合用药对HepG2.A64细胞HBsAg分泌，HBV DNA复制，p38和NTPC mRNA表达，细胞中p38和p-p38蛋白表达水平的影响。结果：与单独用药比较，氧化苦参碱和恩替卡韦联合用药72 h后能显著抑制HBsAg分泌，HBV DNA复制（P<0.05），同时能显著降低NTCP基因和蛋白的表达水平以及磷酸化p38蛋白水平。结论：氧化苦参碱联合恩替卡韦可能通过下调NTCP的表达同时抑制p38蛋白磷酸化达到抗病毒的效果。氧化苦参碱联合恩替卡韦能显著增强抗耐药HBV病毒的效果。

Abstract

Objective: To investigate the anti-hepatitis B virus(HBV) effects of sophora alkaloids combined with entecavir

and observe its effect on p38 MAPK and sodium taurocholote cotransporting poly peptide(NTCP) expression levels. Method: Entecavir-resistant hepatitis HBV replication ell lines HepG2.A64 were used as the models

and then CPE and CCK-8 were used respectively to investigate the cytotoxicity in case of oxymatrine

matrine

sophoridine

sophocarpine or nucleoside drug entecavir as well as combined use of entecavir and any above sophora alkaloids. The levels of HbsAg

HBV DNA replication

p38 and NTCP mRNA expression

as well as p38 and p-p38 protein expression levels were detected by ELISA

Real-time PCR and Western blot. Result: As compared with medication alone

HBsAg secretion

HBV DNA replication were significantly inhibited 72 h after combined use of oxymatrine and entecavir

and NTCP expression levels and p38 phosphorylation levels were significantly reduced. Conclusion: The combined use of oxymatrine and entecavir could achieve the anti-HBV effect by down-regulating the expression of NTCP and inhibiting the phosphorylation of p38.The combined use of oxymatrine and entecavir could significantly increase anti-HBV effect.

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