Sleem Rasha, Salah Ahmed, Alziz Amal Abd, Daif Ahmed A, Megeed Ahmed A Abdel, Khalil Hany
Department of Molecular Biology, Genetic Engineering and Biotechnology Research Institute, University of Sadat City, El-Sadat City, Menoufia Governate, Egypt.
Molecular Diagnostic Department, Genetic Engineering and Biotechnology Research Institute, University of Sadat City, El-Sadat City, Menoufia Governate, Egypt.
Arch Virol. 2025 May 3;170(6):120. doi: 10.1007/s00705-025-06294-6.
Hepatitis C virus (HCV) infection is a significant global health concern, as both acute and chronic hepatitis caused by HCV can lead to liver cancer and long-term liver damage. Thymoquinone (TQ), the active compound found in the remarkable herb Nigella sativa, has various anti-inflammatory and antiproliferative effects. In this study, we investigated the effect of TQ on the interferon-alpha (IFN-α) pathway and its ability to prevent HCV replication in the HepG2 cell line. Our findings showed no significant alterations in cell viability or lactate dehydrogenase (LDH) production in TQ-treated cells, while significant alteration in both factors was detected in cells treated with Sovaldi, the most commonly used drug for treatment of HCV infection. Interestingly, the level of the HCV NS5A protein was significantly reduced in infected cells treated with either TQ or Sovaldi in a dose-dependent manner. The expression of phosphorylated Raf-1 (phospho-Raf-1) and phospho-Mek-1 in infected cells was inhibited by TQ treatment, and the potential interaction between TQ and Ref-1 was confirmed by a molecular docking simulation. Unlike autophagy-related 12 (Atg12), the expression of LC3B in infected cells was also inhibited in a dose-dependent manner by TQ treatment. Conversely, the levels of interleukin-27 (IL-27) and interferon-alpha (IFN-α) in infected cells were significantly increased in a time- and dose-dependent manner by TQ treatment. These data suggest that TQ exerts antiviral effects in HepG2 cells by disrupting HCV replication through targeting of the Raf-1 signaling pathway and promoting the overproduction of IL-27 and IFN-α in infected cells.
丙型肝炎病毒（HCV）感染是一个重大的全球健康问题，因为HCV引起的急性和慢性肝炎都可导致肝癌和长期肝损伤。百里醌（TQ）是著名草药黑种草中发现的活性化合物，具有多种抗炎和抗增殖作用。在本研究中，我们研究了TQ对干扰素-α（IFN-α）途径的影响及其在HepG2细胞系中预防HCV复制的能力。我们的研究结果表明，TQ处理的细胞中细胞活力或乳酸脱氢酶（LDH）产生没有显著变化，而在用治疗HCV感染最常用的药物索磷布韦处理的细胞中，这两个因素都有显著变化。有趣的是，用TQ或索磷布韦处理的感染细胞中，HCV NS5A蛋白水平均以剂量依赖性方式显著降低。TQ处理抑制了感染细胞中磷酸化Raf-1（磷酸-Raf-1）和磷酸-Mek-1的表达，分子对接模拟证实了TQ与Ref-1之间的潜在相互作用。与自噬相关蛋白12（Atg12）不同，TQ处理也以剂量依赖性方式抑制了感染细胞中LC3B的表达。相反，TQ处理以时间和剂量依赖性方式显著增加了感染细胞中白细胞介素-27（IL-27）和干扰素-α（IFN-α）的水平。这些数据表明，TQ通过靶向Raf-1信号通路破坏HCV复制并促进感染细胞中IL-27和IFN-α的过量产生，从而在HepG2细胞中发挥抗病毒作用。
