Epigallocatechin-3-gallate suppresses Hepatitis B virus replication through activating the AMPK/TFEB pathway to promote autophagic degradation of viral core protein

Abstract

Epigallocatechin-3-gallate (EGCG), a major polyphenolic compound in green tea, exhibits anti-viral activity against multiple viruses, including hepatitis B virus (HBV). However, its role in HBV replication and the underlying mechanisms remain incompletely understood. In this study, we investigated the effects of EGCG on HBV replication and its modulation of autophagy using two established HBV cell models. Our results show that EGCG significantly reduces secreted levels of hepatitis B surface antigen (HBsAg) and HBV deoxyribonucleic acid (DNA), as well as intracellular HBV DNA replicative intermediates, encapsidated pregenomic ribonucleic acid (pgRNA), and core protein (HBc), without affecting total HBV messenger RNAs (mRNAs) or pgRNA levels. EGCG enhances autophagic flux, evidenced by increased autophagosome formation and accelerated turnover of the selective autophagy receptor p62 and LC3-Ⅱ. This enhanced autophagy promotes HBc degradation. Pharmacological inhibition of autophagy with 3-methyladenine (3-MA), chloroquine (CQ), or bafilomycin A1 (BafA1) abolished the suppressive effect of EGCG on HBV. Notably, treatment with CQ or BafA1 together with EGCG markedly increased HBV production by blocking autophagic degradation and inducing accumulation of autophagosomes—effects similar to those induced by the autophagy activator rapamycin, which facilitates HBV replication. Mechanistically, EGCG activates the adenosine 5'-monophosphate-activated protein kinase (AMPK)/transcription factor EB (TFEB) signaling axis, leading to enhanced lysosomal biogenesis and ATP production, thereby promoting autophagic clearance. Pharmacological or genetic inhibition of AMPK attenuated TFEB transcriptional activity, suppressed lysosomal biogenesis and ATP generation, impaired autophagic degradation, increased HBc levels, and ultimately enhanced HBV replication. Conversely, pharmacological activation of AMPK produced opposing effects. These findings reveal a novel mechanism by which EGCG inhibits HBV: EGCG promotes autophagic degradation of the viral core protein via activation of the AMPK/TFEB signaling pathway.

Keywords

EGCG / HBV / Autophagy / AMPK / TFEB / Lysosome biogenesis

Funding

This work was supported by the National Natural Science Foundation of China (No. 81600470) and the Natural Science Foundation of Shandong Province of China (Nos. ZR2022MC053 and ZR2023MH122).

Supporting information

Supporting information for this work can be obtained by contacting the corresponding authors via E-mail.

Acknowledgements

We thank Prof. Liu Wei from Zhejiang University School of Medicine for providing us with GFP-LC3 plasmid, GFP-mCherry-LC3 plasmid, GFP-mCherry-p62 plasmid and pcDNA3.0-1.3HBV plasmid. We thank Prof. Chen Zhi from State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine for providing us with HepG2.2.15 cells and HuH-7 cells.

Declaration of competing interest

These authors have no conflict of interest to declare.

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