低密度脂蛋白受体在丙型肝炎病毒感染原代人肝细胞的过程中发挥作用。

The low-density lipoprotein receptor plays a role in the infection of primary human hepatocytes by hepatitis C virus.

作者信息

Molina Sonia, Castet Valérie, Fournier-Wirth Chantal, Pichard-Garcia Lydiane, Avner Rachel, Harats Dror, Roitelman Joseph, Barbaras Ronald, Graber Pierre, Ghersa Paola, Smolarsky Moshe, Funaro Ada, Malavasi Fabio, Larrey Dominique, Coste Joliette, Fabre Jean-Michel, Sa-Cunha Antonio, Maurel Patrick

机构信息

Inserm, U632, Hepatic Physiopathology, 1919 route de Mende, 34293 Montpellier Cedex 5, France.

出版信息

J Hepatol. 2007 Mar;46(3):411-9. doi: 10.1016/j.jhep.2006.09.024. Epub 2006 Nov 13.

DOI:10.1016/j.jhep.2006.09.024

PMID:17156886

Abstract

BACKGROUND/AIMS: The direct implication of low-density lipoprotein receptor (LDLR) in hepatitis C virus (HCV) infection of human hepatocyte has not been demonstrated. Normal primary human hepatocytes infected by serum HCV were used to document this point.

METHODS

Expression and activity of LDLR were assessed by RT-PCR and LDL entry, in the absence or presence of squalestatin or 25-hydroxycholesterol that up- or down-regulates LDLR expression, respectively. Infection was performed in the absence or presence of LDL, HDL, recombinant soluble LDLR peptides encompassing full-length (r-shLDLR4-292) or truncated (r-shLDLR4-166) LDL-binding domain, monoclonal antibodies against r-shLDLR4-292, squalestatin or 25-hydroxycholesterol. Intracellular amounts of replicative and genomic HCV RNA strands used as end point of infection were assessed by RT-PCR.

RESULTS

r-shLDLR4-292, antibodies against r-shLDLR4-292 and LDL inhibited viral RNA accumulation, irrespective of genotype, viral load or liver donor. Inhibition was greatest when r-shLDLR4-292 was present at the time of inoculation and gradually decreased as the delay between inoculation and r-shLDLR4-292 treatment increased. In hepatocytes pre-treated with squalestatin or 25-hydroxycholesterol before infection, viral RNA accumulation increased or decreased in parallel with LDLR mRNA expression and LDL entry.

CONCLUSIONS

LDLR is involved at an early stage in infection of normal human hepatocytes by serum-derived HCV virions.

摘要

背景/目的：低密度脂蛋白受体（LDLR）在丙型肝炎病毒（HCV）感染人肝细胞中的直接作用尚未得到证实。本研究采用血清HCV感染的正常原代人肝细胞来证明这一点。

方法

在存在或不存在分别上调或下调LDLR表达的角鲨烯抑制素或25-羟基胆固醇的情况下，通过逆转录聚合酶链反应（RT-PCR）和低密度脂蛋白（LDL）摄取来评估LDLR的表达和活性。在存在或不存在LDL、高密度脂蛋白（HDL）、包含全长（r-shLDLR4-292）或截短（r-shLDLR4-166）LDL结合域的重组可溶性LDLR肽、抗r-shLDLR4-292单克隆抗体、角鲨烯抑制素或25-羟基胆固醇的情况下进行感染。通过RT-PCR评估用作感染终点的细胞内复制型和基因组HCV RNA链的量。

结果

r-shLDLR4-292、抗r-shLDLR4-292抗体和LDL均可抑制病毒RNA积累，与基因型、病毒载量或肝脏供体无关。当接种时存在r-shLDLR4-292时抑制作用最大，并且随着接种与r-shLDLR4-292处理之间的时间间隔增加，抑制作用逐渐减弱。在感染前用角鲨烯抑制素或25-羟基胆固醇预处理的肝细胞中，病毒RNA积累与LDLR mRNA表达和LDL摄取平行增加或减少。