Liu Zhengyun, Yang Haiyan, Wen Juan, Xiao Dongyan, Yu Wan, Luo Guo, Gong Qihai, Wang Huan
Key Laboratory of Infectious Disease and Biosafety, Provincial Department of Education, Zunyi Medical University, Zunyi, Guizhou, China.
Department of Basic Teaching, Zunyi Medical and Pharmaceutical College, Zunyi, Guizhou, China.
Front Pharmacol. 2025 Apr 1;16:1544714. doi: 10.3389/fphar.2025.1544714. eCollection 2025.
To investigate the potential anti-hepatitis B virus (HBV) activity of Icariside Ⅱ (ICS Ⅱ), and elucidate its underlying mitochondrial dynamics mechanisms.
The study employed and assays to evaluate anti-HBV effects of ICS Ⅱ. An HBV replicating mouse model was established through hydrodynamic injection of pAAV/HBV1.2, the impact of ICS Ⅱ on HBV replication and liver toxicity was assessed. cell-based assays used HBV-positive HepG2.2.15 cells. Cytotoxicity was determined with CCK-8 assay, while ELISA and qPCR were employed to measure HBsAg, HBeAg, and HBV DNA levels. The livers of ICS II-treated HBV-infected mice were taken for transcriptome sequencing to screen for different genes, and the results were verified by Western Blot. Mitochondrial morphology and dynamics were visualized using confocal imaging and transmission electron microscopy. Key protein expressions related to mitochondrial fission and fusion were analyzed via WB. Intracellular ROS generation was assessed using fluorescence staining.
The study found that ICS Ⅱ exhibited significant anti-HBV effects both and . The results of RNA-Seq indicated that ICS Ⅱ modulated the mRNA levels of Fisl, a protein associated with mitochondrial dynamics, during the anti-HBV response. It induced mitochondrial fragmentation and enhanced mitochondrial motility in HBV-positive cells. Notably, key proteins associated with mitochondrial fission and fusion demonstrated alterations favoring fission. Furthermore, ICS Ⅱ effectively reduced ROS production in HBV-positive cells.
ICS Ⅱ exhibits significant anti-HBV potential through its regulation of mitochondrial dynamics and ROS production.
研究淫羊藿苷Ⅱ（ICSⅡ）潜在的抗乙型肝炎病毒（HBV）活性，并阐明其潜在的线粒体动力学机制。
本研究采用[具体方法1]和[具体方法2]实验评估ICSⅡ的抗HBV作用。通过尾静脉注射pAAV/HBV1.2建立HBV复制小鼠模型，评估ICSⅡ对HBV复制和肝毒性的影响。基于细胞的实验使用HBV阳性的HepG2.2.15细胞。用CCK-8法测定细胞毒性，采用ELISA和qPCR检测HBsAg、HBeAg和HBV DNA水平。取接受ICSⅡ治疗的HBV感染小鼠的肝脏进行转录组测序以筛选差异基因，结果通过蛋白质免疫印迹法（Western Blot）验证。使用共聚焦成像和透射电子显微镜观察线粒体形态和动力学。通过蛋白质免疫印迹法分析与线粒体分裂和融合相关的关键蛋白表达。使用荧光染色评估细胞内活性氧（ROS）的产生。
研究发现ICSⅡ在[具体实验1]和[具体实验2]中均表现出显著的抗HBV作用。RNA测序结果表明，在抗HBV反应过程中，ICSⅡ调节了与线粒体动力学相关的蛋白Fisl的mRNA水平。它诱导HBV阳性细胞中的线粒体碎片化并增强线粒体运动性。值得注意的是，与线粒体分裂和融合相关的关键蛋白表现出有利于分裂的变化。此外，ICSⅡ有效降低了HBV阳性细胞中的ROS产生。
ICSⅡ通过调节线粒体动力学和ROS产生表现出显著的抗HBV潜力。
