Targeting HCK in M1 macrophages alleviates postoperative abdominal adhesions by suppressing NF-κB/IL6 axis-driven mesothelial-to-mesenchymal transition

Abstract Postoperative abdominal adhesions (PAAs) are the result of excessive or prolonged inflammation. M1 macrophages are the primary producers of proinflammatory cytokines; however, their direct contribution to PAAs and the underlying mechanism remain unclear. The results of our previous study and public database analysis revealed that peritoneal injury induced M1 macrophage infiltration with specific hematopoietic cell kinase (HCK) upregulation. In vivo HCK inhibition reduced M1 macrophage accumulation in peritoneal injury. Accordingly, in vitro, HCK inhibitors and HCK knockout suppressed M1 macrophage polarization by downregulating the NF-κB pathway. Transwell and FITC-gelatin degradation assay confirmed that HCK deficiency abolished macrophage migration and invasion by disrupting podosome assembly. Furthermore, the causal role of M1 macrophages in PAAs was confirmed by abdominal macrophage depletion and adoptive transfer. M1 macrophage HCK orchestrated the IL6-driven proadhesive mesothelial-to-mesenchymal transition (MMT) via the NF-κB pathway. Finally, perioperative HCK inhibition was verified to alleviate PAA formation through the suppression of MMT-derived myofibroblast transition. Overall, we demonstrated that targeting HCK in M1 macrophages alleviates postoperative abdominal adhesions by suppressing NF-κB/IL6 axis-driven MMT, providing a new target and available strategy for the prevention of PAAs.