Background: Chronic hepatitis B virus (HBV) infection is a well-recognized cause of hepatic injury through prolonged viral replication, inflammation, and oxidative stress. Existing antiviral drugs limit viral replication but cannot eliminate viral transcription or even totally preclude liver injury, thus reemphasizing the significance of drugs with combined antiviral and hepatoprotective effects. Objectives: To evaluate the effects of wedelolactone on HBV replication, gene expression, inflammation, and oxidative stress in an in-vitro model of HBV plasmid transfection with human hepatic cells. Methods: Human hepatocellular carcinoma cells (Huh7) were transfected with a 1.3-mer plasmid and treated with wedelolactone (2.5 - 10 µM). Luciferase assays for HBV promoter activity, Northern blotting and Southern blotting for transcripts and replicative intermediates, qPCR for extracellular HBV DNA, and western blotting for viral antigens such as HBx were performed. Cell cytotoxicity was measured. NF-κB/IκB, inflammatory cytokines (TNF-α, IL-6), and antioxidant markers (Nrf2, HO-1, Keap1) were assessed to evaluate inflammatory and oxidative responses. Results: Wedelolactone significantly suppresses HBV promoter activity, RNAs, core particle formation, and extracellular HBV DNA. It reduced the expression of HBcAg and HBsAg. It inhibited NF-κB activation and cytokine release, while simultaneously enhancing Nrf2/HO-1 signaling, including induction of heme oxygenase-1 by lowering levels of Keap1. Conclusions: Wedelolactone exerts dual antiviral and hepatoprotective actions by inhibiting HBV replication and modulating inflammatory and oxidative stress pathways.

Background: Chronic liver injury progressively induces fibrotic remodeling through persistent inflammatory activity and aberrant extracellular matrix accumulation. Growing evidence indicates that this process involves not only intrahepatic signaling but also gut–liver crosstalk and sirtuin-regulated inflammatory pathways. Hesperetin, a citrus-derived flavanone with reported antioxidant and anti-inflammatory properties, has not been adequately evaluated with respect to microbiota-associated changes and SIRT2-associated signaling during fibrotic liver injury. Objectives: This study investigated whether hesperetin attenuates CCl4-induced liver fibrosis in mice by modulating selected gut microbial populations and suppressing SIRT2-associated inflammatory signaling pathways. Methods: Male mice were randomly assigned to the vehicle, CCl4, and CCl4 + hesperetin groups (n = 5 per group). Liver fibrosis was induced by intraperitoneal administration of CCl4 twice weekly for six weeks, and hesperetin was administered orally at 100 mg/kg/day. Histological, biochemical, inflammatory, SIRT2-associated, and selected gut microbial endpoints were assessed. The expression levels of α-SMA, TGF-β1, TNF-α, IL-6, and SIRT2 were quantitatively analyzed using quantitative polymerase chain reaction (qPCR), Western blotting, and an enzyme-linked immunosorbent assay (ELISA). Levels of selected gut microflora were measured using targeted qPCR for Firmicutes, Bacteroidetes, Akkermansia, and Escherichia coli. Results: CCl4 exposure increased collagen deposition, collagen proportionate area (CPA), and serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, indicating fibrotic remodeling and hepatocellular injury. Hesperetin treatment significantly decreased collagen deposition and CPA and partially normalized ALT and AST activities compared with CCl4 treatment alone. Fibrosis biomarkers, including α-SMA and TGF-β1, and inflammatory cytokines, including TNF-α and IL-6, showed significant decreases in expression at both the mRNA and protein levels. Hesperetin reduced CCl4-associated SIRT2 upregulation and restored acetyl-α-tubulin levels, suggesting attenuation of SIRT2-associated deacetylase activity. Conclusions: These exploratory findings suggest that hesperetin attenuates CCl4-induced liver fibrosis by reducing collagen accumulation and biochemical liver injury, suppressing inflammatory and fibrogenic signaling, modulating selected gut microbial populations, and regulating SIRT2-associated acetylation signaling.