沉默 SIRT1 通过调节 Pol 表达和激活 JAK-STAT 信号通路促进 IFN-α 的抗 HBV 作用。

Silencing SIRT1 promotes the anti-HBV action of IFN-α by regulating Pol expression and activating the JAK-STAT signaling pathway.

机构信息

School of Public Health, North China University of Science and Technology, Tangshan, Hebei Province, China.

School of Life Science, North China University of Science and Technology, Tangshan, Hebei Province, China.

出版信息

Int Immunopharmacol. 2023 Nov;124(Pt B):110939. doi: 10.1016/j.intimp.2023.110939. Epub 2023 Sep 21.

DOI:10.1016/j.intimp.2023.110939

PMID:37741128

Abstract

PURPOSE

The purpose this study is to investigate the impact of SIRT1 on the anti-HBV activity of IFN-α and further elucidate its underlying mechanism.

METHODS

HepG2.2.15 cells stably transfected with HBV virus were chosen as the primary study subject. IFN-α was used to stimulate the cells and regulate the expression of SIRT1, and the JAK-STAT pathway and HBV-related indices were measured by qRT-PCR, Western blotting and ELISA. Immunofluorescence (IF) was used to detect the nuclear translocation of STAT1 and STAT2. Coimmunoprecipitation (Co-IP) was used to detect the binding of SIRT1 to HBV Polymerase (Pol).

RESULTS

In HepG2.2.15 cells, we found changes in SIRT1 expression. We show that silencing SIRT1 promotes the IFN-α-triggered Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway and consequently enhances the antiviral effects of IFN-α against HBV replication. Importantly, SIRT1 can interact with Pol and increase JAK-STAT activity by regulating Pol expression. Additionally, the inhibition of SIRT1 activity by treatment with the SIRT1 inhibitor selisistat enhanced the anti-HBV effect of IFN-α and JAK-STAT pathway activity.

CONCLUSION

In conclusion, our results demonstrate that silencing SIRT1 activates the JAK-STAT pathway and enhances the anti-HBV activity of IFN-α by inhibiting Pol expression. This would be a promising therapeutic target to improve the efficacy of IFN-α in the treatment of CHB.

摘要

目的

本研究旨在探讨 SIRT1 对 IFN-α抗 HBV 活性的影响，并进一步阐明其潜在机制。

方法

选择稳定转染 HBV 病毒的 HepG2.2.15 细胞作为主要研究对象。用 IFN-α刺激细胞，调节 SIRT1 的表达，通过 qRT-PCR、Western blot 和 ELISA 检测 JAK-STAT 通路和 HBV 相关指标。免疫荧光（IF）检测 STAT1 和 STAT2 的核转位。免疫共沉淀（Co-IP）检测 SIRT1 与 HBV 聚合酶（Pol）的结合。

结果

在 HepG2.2.15 细胞中，我们发现 SIRT1 表达发生变化。我们表明，沉默 SIRT1 可促进 IFN-α 触发的 Janus 激酶-信号转导和转录激活因子（JAK-STAT）信号通路，从而增强 IFN-α 对 HBV 复制的抗病毒作用。重要的是，SIRT1 可以通过调节 Pol 表达与 Pol 相互作用并增加 JAK-STAT 活性。此外，用 SIRT1 抑制剂 selisistat 处理抑制 SIRT1 活性增强了 IFN-α 的抗 HBV 作用和 JAK-STAT 通路活性。