Lysine acetyltransferase 2A-mediated succinylation of adenosine monophosphate-activated protein kinase suppresses gallstone formation by inhibiting inflammation and pyroptosis.

BACKGROUND: Cholelithiasis is a prevalent biliary tract disorder primarily characterized by gallbladder or biliary stone formation. Although succinylation has been extensively studied as a protein post-translational modification, its role in cholelithiasis remains unexplored. AIM: To investigate the functional role of succinylation in cholelithiasis and determine its underlying molecular mechanisms. METHODS: A murine cholelithiasis model was established through high-fat diet feeding, followed by isolation of mouse gallbladder mucosal epithelial cells (GMECs) for in vitro analysis. Gallbladder tissues and serum samples were collected for subsequent analysis. Inflammatory cytokine production was quantified using enzyme-linked immunosorbent assay. Pyroptosis was analyzed by flow cytometry, while succinylation- and pyroptosis-related protein expression was detected via western blot. RESULTS: Our findings demonstrated that lysine acetyltransferase 2A (KAT2A)-mediated succinylation regulated gallstone formation. KAT2A overexpression inhibited the pyroptosis, inflammatory responses, and promoted the activation of the adenosine monophosphate-activated protein kinase (AMPK)/silent information regulator 1 (SIRT1) signaling pathway in GMECs. Mechanistically, AMPK exhibited succinylation at lysine 170 (K170). Notably, AMPK inhibition significantly increased pyroptosis rates, inflammatory responses, and pyroptosis-related protein expression in GMECs. Furthermore, in vivo experiments revealed that KAT2A overexpression suppressed both inflammation and gallstone formation. CONCLUSION: KAT2A-mediated succinylation of AMPK inhibited cholelithiasis progression by modulating the AMPK/SIRT1 signaling pathway, offering potential therapeutic strategies for this condition.