TET3 Regulates Hepatic Fibrosis in Mice through the Pyroptotic Pathway

Abstract

Background: Hepatic fibrosis (HF) is a prevalent precursor to cirrhosis in various chronic liver diseases, underscoring the need for comprehensive investigations into its etiology, prevention, and management. Pyroptosis, an emerging form of cellular death, is prevalent in liver diseases. Recent research suggests that ten-eleven-translocation protein 3 (TET3), an enzyme responsible for DNA demethylation, plays a vital role in regulating HF, although the precise mechanism remains unclear. This experimental study aimed to investigate the impact of the TET3-mediated pyroptotic pathway on HF in mice and elucidate the underlying regulatory mechanisms. Methods: HF was induced in mice through intraperitoneal injection of carbon tetrachloride (CCl₄), while lipopolysaccharide (LPS) induced pyroptosis in murine hepatocytes. Protein expression was detected using the Western blotting technique, and pathological alterations were evaluated through hematoxylin and eosin (H&E) and Masson staining. Immunofluorescence and dot blotting assessed the expression of 5-hydroxymethylcytosine (5-hmC), while transmission electron microscope measured hepatocyte ultrastructure. The relationship between TET3 and NLR family pyrin domain containing 3 (NLRP3), Caspase-1, and gasdermin D (GSDMD) was evaluated using chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR). Results: The expression of pyroptotic proteins showed a positive correlation with the extent of HF (p < 0.05), while the expression of TET3 and 5-hmC exhibited a negative correlation (p < 0.05). TET3 overexpression successfully prevented HF, upregulated 5-hmC expression, and attenuated pyroptosis (p < 0.05). TET3 administration significantly increased LPS-induced pyroptosis in hepatocytes of mice (p < 0.05). Finally, TET3 was shown to target NLRP3, Caspase-1, and GSDMD genes (p < 0.01). Conclusions: By reversing the classical pathway of pyroptosis, TET3 can potentially improve HF progression in mice by targeting NLRP3, Caspase-1, and GSDMD.