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中华实验和临床感染病杂志(电子版)

中华实验和临床感染病杂志(电子版) ›› 2018, Vol. 12 ›› Issue (06) : 547 -552. doi: 10.3877/cma.j.issn.1674-1358.2018.06.006

所属专题： 文献；

论著

乙型肝炎病毒表面抗原主蛋白与烯酰辅酶A水合酶相互作用对肝细胞胆固醇代谢的影响
张凯1, 姚雯1, 林苏1, 王明芳1, 朱月永1,†()   
  1. 1. 350005　福州市，福建医科大学附属第一医院肝病中心
  • 收稿日期:2018-05-05 出版日期:2018-12-15
  • 通信作者: 朱月永
  • 基金资助:
    福建省自然科学基金项目(No. 2016Y0040); 福建省中青年教师教育科研项目(No. JA15207)

Impact of interactions between small hepatitis B virus surface antigen and enoyl co-enzyme A hydratase on cholesterol metabolism in hepatocytes

Kai Zhang1, Wen Yao1, Su Lin1, Mingfang Wang1, Yueyong Zhu1,†()   

  1. 1. Liver Research Center, The First Affiliated Hospital of Fujian Medical University, Fuzhou 350005, China
  • Received:2018-05-05 Published:2018-12-15
  • Corresponding author: Yueyong Zhu
  • About author:
    Corresponding author: Zhu Yueyong, Email:
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张凯, 姚雯, 林苏, 王明芳, 朱月永. 乙型肝炎病毒表面抗原主蛋白与烯酰辅酶A水合酶相互作用对肝细胞胆固醇代谢的影响[J]. 中华实验和临床感染病杂志(电子版), 2018, 12(06): 547-552.

Kai Zhang, Wen Yao, Su Lin, Mingfang Wang, Yueyong Zhu. Impact of interactions between small hepatitis B virus surface antigen and enoyl co-enzyme A hydratase on cholesterol metabolism in hepatocytes[J]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2018, 12(06): 547-552.

目的

探究乙型肝炎病毒表面抗原主蛋白（SHBs）与烯酰辅酶A水合酶（ECHS1）相互作用对肝细胞胆固醇代谢的影响。

方法

通过在人肝癌细胞株HepG2中瞬时转染SHBs，建立SHBs瞬时转染细胞模型，分别设置空白对照组、空质粒转染组、单纯SHBs转染组、SHBs +阴性序列对照组、ECHS1过表达组以及ECHS1干扰组。在转染后48 h检测ECHS1及载脂蛋白A1（ApoA1）基因的表达和各转染组转氨酶水平及细胞内总胆固醇（TC）含量。

结果

与空白组相比，单纯SHBs转染组在核酸水平ECHS1、ApoA1的表达均下降（P均< 0.001），蛋白水平表达趋势同核酸一致（P < 0.001、P = 0.0025），细胞内TC水平升高（P < 0.001），转氨酶水平升高（PALT < 0.001、PAST < 0.001）。较单纯SHBs转染组，ECHS1过表达组可在核酸和蛋白水平下调ApoA1（P = 0.0072、P < 0.001），且细胞内TC水平下降（P < 0.001），转氨酶水平降低（PALT < 0.001、PAST = 0.0049）。与单纯SHBs转染组相比较，ECHS1干扰组ApoA1核酸和蛋白水平亦降低（P = 0.0096、P < 0.001），TC水平降低（P < 0.001），ALT水平无显著变化（P = 0.28），AST水平升高（P = 0.0069）。

结论

SHBs能够通过下调ApoA1表达来提高肝细胞TC水平，同时ECHS1通过相关信号通路及分子机制抑制TC的生成。

关键词: 肝炎病毒表面抗原主蛋白，乙型, 烯酰辅酶A水合酶, HepG2细胞, 胆固醇代谢
Objective

To investigate the interactions between small hepatitis B virus surface protein (SHBs) and enoyl CoA hydratase short chain 1 (ECHS1) on cholesterol metabolism in hepatocytes.

Methods

Human hepatocellular carcinoma HepG2 cells were transfected with SHBs transiently, and then assigned into the blank control group, the blank plasmid transfection group, the SHBs transfection group, the SHBs + negative sequence control group, the ECHS1 overexpression group and the ECHS1 interference group. The expression of ECHS1 and apolipoprotein A1 (ApoA1) were detected, and the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and intracellular total cholesterol (TC) were measured 48 hours post-transfection.

Results

Significantly lower ECHS1 and ApoA1 expression were detected in the SHBs transfection group than those of the blank control group at both translational (both P < 0.001) and transcriptional levels (P < 0.001, P = 0.0025), and higher intracellular TC (P < 0.001) and serum ALT (P < 0.001) and AST levels (P < 0.001) were measured in SHBs transfection group than those of the blank control group. In addition, lower ApoA1 protein (P = 0.0072) and mRNA expression (P < 0.001), lower intracellular TC levels (P < 0.001), lower serum ALT (P < 0.001) and AST concentrations levels (P = 0.0049) were found in the ECHS1 overexpression group than those in the SHBs transfection group; meanwhile, compared with SHBs transfection group, there were significantly lower ApoA1 nucleic acid and protein levels (P = 0.0096, P < 0.001), decreased TC level (P < 0.001), no significant change of ALT level (P = 0.28), and increased AST level (P = 0.0069) in ECHS1 interference group.

Conclusions

SHBs increases TC levels through downregulating ApoA1 expression in hepatocytes, and ECHS1 may suppress TC production through some signal pathways and molecular mechanism.

Key words: Small hepatitis B virus surface antigen, Enoyl co-enzyme A hydratase, HepG2 cell, Cholesterol metabolism
图1 质粒pcDNA3.1（+）-SHBs双酶切与质粒pcDNA3.1（+）-ECHS1双酶切电泳图及转染细胞中SHBs基因mRNA的表达
图2 不同转染组HepG2细胞中APOA1 mRNA和蛋白表达水平
图3 不同转染组细胞转染48 h后细胞脂质水平
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