基础医学与临床 ›› 2025, Vol. 45 ›› Issue (4): 465-470.doi: 10.16352/j.issn.1001-6325.2025.04.0465

• 研究论文 • 上一篇    下一篇

大蒜素通过HBV启动子SP2抑制HBV复制

卢莉莉, 朱席琳, 伍晓盼*, 刘英*   

  1. 中国医学科学院基础医学研究所 北京协和医学院基础学院 生物化学与分子生物学系, 北京 100005
  • 收稿日期:2024-12-20 修回日期:2025-02-20 出版日期:2025-04-05 发布日期:2025-03-24
  • 通讯作者: *wuxiaopan@ibms.cams.cn; liuyingpumc@163.com
  • 基金资助:
    国家重点研发计划(2021YFC2700601)

Allicin inhibits HBV replication through the HBV promoter SP2

LU Lili, ZHU Xilin, WU Xiaopan*, LIU Ying*   

  1. Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005, China
  • Received:2024-12-20 Revised:2025-02-20 Online:2025-04-05 Published:2025-03-24

摘要: 目的 探究大蒜素(allicin)对乙型肝炎病毒(HBV)复制的影响并初步阐明其调控分子机制。方法 不同浓度的大蒜素处理HepG2.2.15细胞,采用ELISA检测HBsAg和HBeAg的表达水平,CCK-8实验评估大蒜素对细胞活力的影响,从中筛选出适宜的药物浓度;以该浓度的大蒜素处理HepG2-NTCP细胞,48 h后通过RT-qPCR检测HBV相关标志物的表达;通过双荧光素酶报告基因实验,分析4种HBV启动子(Enh Ⅰ/Xp, SP1, SP2 和 CP)的活性,并评估大蒜素对这些启动子活性的影响;使用工具Gene-regulation预测可能与启动子结合的转录因子,过表达转录因子后处理细胞以检测大蒜素对启动子活性的影响。最后,通过ChIP实验验证转录因子与HBV启动子的结合情况,以及大蒜素处理是否改变这种结合。结果 大蒜素呈浓度依赖性显著降低HBsAg表达水平,同时对HBeAg表达有轻微抑制作用(P<0.001);40 μmol/L的大蒜素表现出最佳的抗病毒效果,且对细胞活力无显著影响;此外,40 μmol/L的大蒜素还能显著抑制HBV的DNA复制和转录水平(P<0.05);大蒜素显著抑制了HBV启动子SP2的活性(P<0.001);进一步研究发现,转录因子SP1能够与HBV启动子SP2的DNA序列结合,经大蒜素处理后,该结合显著受到抑制(P<0.001)。结论 大蒜素通过抑制转录因子SP1与HBV启动子SP2的结合,降低HBV的转录水平,进而抑制病毒的复制。

关键词: 大蒜素, 乙型肝炎病毒, 抗病毒, 转录调控, HBV启动子SP2

Abstract: Objective To investigate the effect of allicin on the replication of hepatitis B virus (HBV) and to preliminarily elucidate its underlying molecular mechanisms. Methods HepG 2.2.15 cells were treated with different concentrations of allicin and the levels of HBsAg and HBeAg were assessed by ELISA. Cell viability was evaluated using the CCK-8 assay to determine the optimal concentration of allicin; HepG2-NTCP cells were incubated with the optimal concentration of allicin for 48 hours, and the expression of HBV-related markers was detected by RT-qPCR; The activity of four HBV promoters (Enh Ⅰ/Xp, SP1, SP2, and CP) was analyzed using a dual-luciferase reporter gene experiment. The effect of allicin on promoter activity was assessed; Gene-regulation tools were used to predict potential transcription factor that might bind to the promoter. After over-expressing the transcription factor, cells were incubated with allicin and the effect on promoter activity was examined; Finally, ChIP was used to confirm whether these transcription factors bind to the HBV promoters and whether allicin treatment affects this binding. Results Allicin significantly reduced the expression of HBsAg and slightly lowered the expression of HBeAg(P<0.001); A concentration of 40 μmol/L allicin significantly inhibited HBV DNA replication and transcription(P<0.05), without affecting cell viability; Allicin also significantly suppressed the activity of the HBV promoter SP2(P<0.001). Further investigation revealed that the transcription factor SP1 could bind to the DNA sequence of the HBV promoter SP2, and this binding was significantly inhibited after allicin treatment(P<0.001). Conclusions Allicin inhibits the binding of the transcription factor SP1 to HBV promoter SP2, thereby reducing the transcriptional activity of HBV and suppressing viral replication.

Key words: allicin, Hepatitis B virus, antiviral, transcriptional regulation, HBV promoter SP2

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