An in vitro model of hepatitis C virus genotype 3a-associated triglycerides accumulation
An in vitro model of hepatitis C virus genotype 3a-associated triglycerides accumulation
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DOI：
10.1016/j.jhep.2004.12.034
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发表时间：
2005-05-01
影响因子：
25.7
通讯作者：
Negro, F
中科院分区：
文献类型：
--
作者：
Abid, K;Pazienza, V;Negro, F
背景/目的：丙型肝炎病毒（HCV）在体内和体外诱导脂质积累。尽管临床观察结果与HCV基因型3a对脂质代谢的直接影响一致，但实验系统主要关注基因型1的HCV蛋白的表达。为了扩展这些观察结果，我们建立了表达不同基因型HCV核心的体外模型。方法：将重度（基因型为3a）和非重度（基因型为1b、2a、3h、4h和5a）肝脂肪变性患者的HCV核心蛋白在Huh7细胞中表达。转染48 h后检测核心蛋白表达（免疫组织化学和免疫印迹法）和甘油三酯积累（油红0染色和酶促法）。结果：虽然基因型1b、3a和3h发生甘油三酯积累，但基因型3a核心蛋白表达导致的积累水平最高（即相对于1b约为3倍，相对于3h约为2倍）。这种影响与核心蛋白表达水平无关，在无脂培养基中培养细胞可以消除这种影响。结论：与在慢性丙型肝炎患者中的观察结果一致，HCV基因型3a核心蛋白的体外表达是研究HCV相关脂肪变性机制的理想候选模型。(c) 2005年欧洲肝脏研究协会。Elsevier B.V.版权所有。
Background/Aims: The hepatitis C virus (HCV) induces lipid accumulation in vitro and in vivo. Although clinical observations are consistent with a direct effect of HCV genotype 3a on lipid metabolism, experimental systems have focused on the expression of HCV proteins of genotype 1. To extend these observations, we established an in vitro model expressing the HCV core of different genotypes.Methods: The HCV core protein from patients with severe (genotype 3a) or no (genotypes 1b, 2a, 3h, 4h and 5a) liver steatosis was expressed in Huh7 cells. Core protein expression (by immunohistochemistry and immunoblot) and triglycerides accumulation (by Oil Red 0 stain and enzymatic measurement) were evaluated 48 h after transfection.Results: Although triglyceride accumulation occurred with genotypes 1b, 3a and 3h, the genotype 3a core protein expression resulted in the highest level of accumulation (i.e. about 3-fold with respect to 1b, and 2-fold with respect to 3h). This effect was not related to core protein expression levels and was abolished by culturing cells in lipid-free medium.Conclusions: Consistent with observations in chronic hepatitis C patients, the in vitro expression of HCV genotype 3a core protein is the ideal candidate model for studying the mechanisms of HCV-associated steatosis. (c) 2005 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.