Hk2 Promotes Postburn Intestinal Injury by Facilitating Pyroptosis in Intestinal Epithelial Cells Through Enhancing Gsdmc2 Lactylation.

Abstract

BACKGROUND AND AIM: Intestinal injury is a common complication following burn injury, increasing patient adverse outcomes. Hk2, a key glycolysis gene, promotes lactylation by raising intracellular lactate levels. While pyroptosis is crucial for intestinal homeostasis, its mediation by Hk2-induced lactylation remains unclear. This study elucidates how Hk2 regulates postburn intestinal injury via pyroptosis modulation. METHODS: BALB/c mice were exposed to a boiling water bath to induce a mouse burn model. Mouse intestinal epithelial cells were treated with lipopolysaccharide (LPS) to simulate intestinal injury in vitro. The role of Hk2 on LPS-induced mouse intestinal epithelial cells was evaluated by detecting cell viability, lactate dehydrogenase release, levels of inflammation, and pyroptosis factors and pyroptosis rate. The underlying mechanism was determined by quantitative real-time PCR, coimmunoprecipitation, and IP. Intestinal injury was evaluated by hematoxylin and eosin staining and measurements of inflammation factors. RESULTS: LPS promoted glycolysis and upregulated Hk2 in both models. Increased inflammation, pyroptosis, glycolysis, and histone lactylation caused by LPS were inhibited by Hk2 knockdown but enhanced by Hk2 overexpression. Exogenous addition of lactate reversed the inhibition of Hk2 knockdown on pyroptosis and inflammation in LPS-induced mouse intestinal epithelial cells. Mechanistically, Hk2 knockdown reduced the stability of the Gsdmc2 protein by decreasing its lactylation. Moreover, Hk2 knockdown improved survival rate, pathological changes, and inflammation of the intestine and downregulated Gsdmc2 in the mouse burn model. CONCLUSION: Hk2 knockdown mitigated postburn intestinal injury by inhibiting pyroptosis through decreased Gsdmc2 lactylation, providing a theoretical basis for developing clinical treatment strategies for this condition.