Vaccine and Infectious Disease Organization, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
J Gen Virol. 2010 Jun;91(Pt 6):1388-95. doi: 10.1099/vir.0.017418-0. Epub 2010 Feb 3.
Hepatitis C virus genotype-3a (HCV-3a) is directly linked to the development of steatosis. We previously showed that, through sterol regulatory element binding protein-1 (SREBP-1), HCV-3a core protein upregulates the promoter activity of fatty acid synthase, a major enzyme involved in de novo lipid synthesis. In this study, we investigated whether HCV-3a core can activate SREBP-1 and studied the role of phosphoinositide 3-kinase (PI3K)-Akt-2 pathway in modulating SREBP-1 activity by HCV-3a core. To determine whether HCV-3a core could activate SREBP-1, the level of mature SREBP-1 was analysed by Western blotting. Our results showed that the level of mature SREBP-1 was enhanced by HCV-3a core protein after transient expression and in the chimeric HCV-3a core/1b replicon cells in comparison to controls. To investigate the role of the PI3K-Akt-2 pathway in SREBP-1 activation by HCV-3a core, PI3K and Akt-2 activity was inhibited by using the chemical inhibitor LY294002, a dominant-negative Akt-2 plasmid, or knockdown of Akt-2 by small hairpin RNA. Our results showed that inhibition of PI3K and Akt-2 was associated with reduced SREBP-1 activation by HCV-3a core. These results indicate a role for PI3K and Akt-2 in increasing SREBP-1 activity by HCV-3a core protein and provide a mechanism of steatosis caused by HCV.
丙型肝炎病毒基因型 3a(HCV-3a)与脂肪变性的发生直接相关。我们之前的研究表明,HCV-3a 核心蛋白通过固醇调节元件结合蛋白-1(SREBP-1)上调脂肪酸合成酶的启动子活性,该酶是参与从头脂质合成的主要酶。在这项研究中,我们研究了 HCV-3a 核心蛋白是否可以激活 SREBP-1,并研究了磷酸肌醇 3-激酶(PI3K)-Akt-2 通路在调节 HCV-3a 核心蛋白激活 SREBP-1中的作用。为了确定 HCV-3a 核心蛋白是否可以激活 SREBP-1,我们通过 Western blot 分析成熟 SREBP-1 的水平。结果表明,与对照组相比,HCV-3a 核心蛋白瞬时表达和嵌合 HCV-3a 核心/1b 复制子细胞中成熟 SREBP-1 的水平增强。为了研究 PI3K-Akt-2 通路在 HCV-3a 核心蛋白激活 SREBP-1中的作用,我们使用化学抑制剂 LY294002、显性失活 Akt-2 质粒或 Akt-2 的小发夹 RNA 敲低抑制 PI3K 和 Akt-2 的活性。结果表明,PI3K 和 Akt-2 的抑制与 HCV-3a 核心蛋白激活 SREBP-1减少相关。这些结果表明 PI3K 和 Akt-2 在 HCV-3a 核心蛋白增加 SREBP-1活性中起作用,并为 HCV 引起的脂肪变性提供了一种机制。