Hepatoprotective Effects of Imeglimin Are Associated With Preserved Mitochondrial Bioenergetics and Reduced Oxidative Stress in a Murine Cholestasis Model.

Abstract

Purpose: Biliary atresia is a cholestatic liver disease that causes persistent inflammation and reduces the hepatic reserve. New therapeutic concepts are crucial to improve native liver survival. We investigated whether imeglimin, a mitochondria-targeting agent, has hepatoprotective effects on cholestatic liver disease using a murine bile duct ligation (BDL) model.

Methods: BDL was performed in 4-5-week-old mice. After BDL, the mice received intraperitoneal imeglimin or vehicle injections. Sham-operated mice received vehicle. Blood and liver samples were collected on days 7 and 14 for biochemical and histological analysis. Mitochondrial morphology was examined by transmission electron microscopy. Quantitative PCR and immunoblotting were performed for inflammation and mitochondrial stress markers. Metabolomic profiling was conducted using capillary electrophoresis-mass spectrometry. Transcriptome was analyzed by RNA sequencing. Seahorse assays, ATP quantification, and MitoSOX staining were used to evaluate mitochondrial function and oxidative stress.

Results: Imeglimin improved survival after BDL, and reduced serum AST, ALT, and bilirubin levels. Tissue fibrosis and Col1a1 and Acta2 expression were attenuated in imeglimin-treated livers. Glycolysis, amino acid metabolism, and mitochondrial electron transport activity were enhanced. Expression of genes associated with mitochondrial function, antioxidant defense, and metabolic regulation was upregulated. Mitochondrial morphology was preserved, and mitochondrial number and size increased. Imeglimin elevated oxygen consumption, extracellular acidification rates, and ATP levels of BDL mice livers, while reducing mitochondrial superoxide and oxidative protein modification by 4-hydroxynonenal. Expression levels of Il1β, Il6, and Tnf decreased.

Conclusion: Imeglimin exerted hepatoprotective effects on BDL-operated mice, which are associated with preserved mitochondrial bioenergetic function and reduced oxidative stress.