MiR-324-5p mitigates experimental acute pancreatitis by suppressing Rock2 expression.

Abstract

Acute pancreatitis (AP) is a lethal pancreatic disease with limited therapeutic options. MicroRNA-324-5p (miR-324-5p) has shown dual roles across diseases, but its specific role and molecular targets in AP remain unclear. Plasma miR-324-5p levels were measured in 80 AP patients and 40 healthy controls using RT-qPCR. An in vivo AP mouse model was induced by caerulein, and agomir miR-324-5p was administered to assess its therapeutic effect. In vitro, MPC-83 pancreatic acinar cells were treated with caerulein and transfected with miR-324-5p mimics or co-transfected with Rock2 overexpression plasmids. Apoptosis and inflammation were evaluated using qPCR, Western blot, ELISA, and histology. Potential targets of miR-324-5p were predicted using StarBase, microT, miRmap, and PITA databases. Rock2 targeting was experimentally confirmed using dual-luciferase reporter assay, RNA pull-down, and RIP assays. MiR-324-5p was significantly downregulated in AP patient plasma and AP mouse pancreas. Experimental agomir-mediated overexpression of miR-324-5p alleviated pancreatic injury alleviated pancreatic injury, reduced serum amylase/lipase, MPO levels, and suppressed cytokines expression in vivo. In vitro, miR-324-5p restored cell viability, inhibited apoptosis, and suppressed inflammatory cytokines in caerulein-treated MPC-83 cells. Bioinformatic and experimental assays identified Rock2 as a direct target of miR-324-5p. Rock2 overexpression reversed the protective effects of miR-324-5p on apoptosis and inflammation in both cell and animal models. MiR-324-5p plays a protective role in AP by directly targeting Rock2 and suppressing inflammatory and apoptotic responses.