Deguelin inhibits HCV replication through suppressing cellular autophagy via down regulation of Beclin1 expression in human hepatoma cells
Deguelin inhibits HCV replication through suppressing cellular autophagy via down regulation of Beclin1 expression in human hepatoma cells
复制标题
Deguelin 通过下调人肝癌细胞中 Beclin1 的表达来抑制细胞自噬，从而抑制 HCV 复制
DOI：
10.1016/j.antiviral.2020.104704
复制
发表时间：
2020-02-01
影响因子：
7.6
通讯作者：
Ye, Li
中科院分区：
文献类型：
--
作者：
Liao, Weibo;Liu, Xin;Ye, Li
目的：Deguelin 是一种源自 Mundulea sericea（豆科）和其他一些植物的天然化合物，具有抑制自噬的活性，自噬是丙型肝炎病毒 (HCV) 复制所需的细胞机制。本研究旨在阐明德桂林对HCV复制的影响及其机制。方法：采用HCV JFH-1-Huh7感染系统进行研究。使用实时RT-PCR、蛋白质印迹、荧光显微镜检测来测量病毒或细胞因子的表达水平。采用过表达和沉默表达技术来确定关键细胞因子的作用。结果：Deguelin处理Huh7细胞以剂量和时间依赖性方式显着抑制HCV JFH-1复制。与对照细胞相比，Deguelin 处理抑制了 Huh7 细胞中的自噬，表现为 LC3B-II 水平降低、LC3B-I 向 LC3B-II 的转化、GFP-LC3 斑点的形成以及 p62 水平增加。 HCV 感染可诱导自噬，而鱼胶素治疗也可抑制自噬。机制研究表明，deguelin 抑制 Beclinl 的表达，而 Beclinl 是自噬中启动自噬体形成的关键细胞因子。经deguelin处理的Huh7细胞中Beclinl的过表达或沉默表达可分别减弱或增强deguelin对自噬的抑制作用，从而相应地部分恢复或进一步抑制HCV复制。结论：Deguelin 可能通过其对自噬的抑制作用而成为一种新型抗 HCV 化合物，值得进一步研究作为 HCV 感染的潜在治疗剂。
Aims: Deguelin, a natural compound derived from Mundulea sericea (Leguminosae) and some other plants exhibits an activity to inhibit autophagy, a cellular machinery required for hepatitis C virus (HCV) replication. This study aimed to illuminate the impact of deguelin on HCV replication and mechanism(s) involved.Methods: HCV JFH-1-Huh7 infectious system was used for the investigation. Real time RT-PCR, Western blot, fluorescent microscopy assay were used to measure the expression levels of viral or cellular factors. Overexpression and silencing expression techniques were used to determine the role of key cellular factors.Results: Deguelin treatment of Huh7 cells significantly inhibited HCV JFH-1 replication in a dose- and time-dependent manner. Deguelin treatment suppressed autophagy in Huh7 cells, evidenced by the decrease of LC3B-II levels, the conversion of LC3B-I to LC3B-II, and the formation of GFP-LC3 puncta as well as the increase of p62 level in deguelin-treated cells compared with control cells. HCV infection could induce autophagy which was also suppressed by deguelin treatment. Mechanism research reveals that deguelin inhibited expression of Beclinl, which is a key cellular factor for the initiation of the autophagosome formation in autophagy. Overexpression or silencing expression of Beclinl in deguelin-treated Huh7 cells could weaken or enhance the inhibitory effect on autophagy by deguelin, respectively, and thus partially recover or further inhibit HCV replication correspondingly. Conclusions: Deguelin may serve as a novel anti-HCV compound via its inhibitory effect on autophagy, which warrants further investigation as a potential therapeutic agent for HCV infection.