Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan.
Teijin Institute for Bio-medical Research, Teijin Pharma Ltd., Tokyo, Japan.
Antiviral Res. 2018 Dec;160:55-63. doi: 10.1016/j.antiviral.2018.10.014. Epub 2018 Oct 16.
Supplementation with vitamin D (VD) has been reported to improve the efficacy of interferon-based therapy for chronic hepatitis C. We found that 25-hydroxyvitamin D (25-(OH)D), one of the metabolites of VD, has antiviral effects by inhibiting the infectious virus production of the hepatitis C virus (HCV). In this study, to clarify the underlying mechanisms of the anti-HCV effects, we searched VD derivatives that have anti-HCV effects and identified the common target molecule in the HCV life cycle by using an HCV cell culture system. After infection of Huh-7.5.1 cells with cell culture-generated HCV, VD derivatives were added to culture media, and the propagation of HCV was assessed by measuring the HCV core antigen levels in culture media and cell lysates. To determine the step in the HCV life cycle affected by these compounds, the single-cycle virus production assay was used with a CD81-negative cell line. Of the 14 structural derivatives of VD, an anti-HCV effect was detected in 9 compounds. Cell viability was not affected by these effective compounds. The 2 representative VD derivatives inhibited the infectious virus production in the single-cycle virus production assay. Treatment with these compounds and 25-(OH)D suppressed the expression of apolipoprotein A1 and C3, which are known to be involved in infectious virus production of HCV, and the knockdown of these apolipoproteins reduced infectious virus production. In conclusion, we identified several compounds with anti-HCV activity by screening VD derivatives. These compounds reduce the infectious virus production of HCV by suppressing the expression of apolipoproteins in host cells.
补充维生素 D（VD）已被报道可提高基于干扰素的慢性丙型肝炎治疗的疗效。我们发现 VD 的一种代谢产物 25-羟维生素 D（25-(OH)D）通过抑制丙型肝炎病毒（HCV）的感染性病毒产生具有抗病毒作用。在这项研究中，为了阐明抗 HCV 作用的潜在机制，我们搜索了具有抗 HCV 作用的 VD 衍生物，并通过 HCV 细胞培养系统鉴定了 HCV 生命周期中的共同靶分子。在 Huh-7.5.1 细胞中用细胞培养产生的 HCV 感染后，将 VD 衍生物添加到培养基中，并通过测量培养基和细胞裂解物中的 HCV 核心抗原水平来评估 HCV 的繁殖。为了确定这些化合物影响 HCV 生命周期的步骤，使用 CD81 阴性细胞系进行了单循环病毒产生测定。在 14 种 VD 的结构衍生物中，有 9 种化合物具有抗 HCV 作用。这些有效化合物对细胞活力没有影响。这 2 种代表性的 VD 衍生物在单循环病毒产生测定中抑制了感染性病毒的产生。用这些化合物和 25-(OH)D 处理可抑制载脂蛋白 A1 和 C3 的表达，已知它们参与 HCV 的感染性病毒产生，而这些载脂蛋白的敲低可减少感染性病毒的产生。总之，我们通过筛选 VD 衍生物鉴定了几种具有抗 HCV 活性的化合物。这些化合物通过抑制宿主细胞中载脂蛋白的表达来减少 HCV 的感染性病毒产生。
