Abstract
<title>Abstract</title> <p>Background Acetaminophen (APAP) overdose is the primary cause of acute liver failure (ALF). Its pathological mechanism involves multiple types of programmed cell death, but whether a correlation exists between these programmed death processes remains unclear. Mesenchymal stem cell-derived apoptotic vesicles (MSC-ApoVs) possess excellent immunomodulatory potential, but their regulatory role in APAP-ALF is not yet defined. Materials and Methods This study utilized an APAP-induced mouse ALF model and a hepatocyte injury model to evaluate the therapeutic effects of MSC-ApoVs. Through transcriptome sequencing (RNA-seq), miRNA sequencing, and molecular biology experiments, the specific molecular mechanism of MSC-ApoVs in regulating PANoptosis was explored. Results The study found that multiple forms of programmed death exist in APAP-induced liver injury, and PANoptosis occurs in hepatocytes. MSC-ApoVs can significantly reduce serum transaminase levels in APAP mice, decrease liver tissue necrosis, and inhibit the expression of pyroptosis-, apoptosis-, and necroptosis-related proteins in hepatocytes, successfully blocking the assembly of the RIPK3-ASC-Caspase-8 PANoptosome complex. Mechanistically, RNA-seq screened out NLRC5 as a key target; knocking down NLRC5 can mimic the anti-PANoptotic and anti-inflammatory effects of MSC-ApoVs. Further research confirmed that MSC-ApoVs are rich in miR-378a-5p, which directly binds to and inhibits NLRC5 mRNA. Utilizing engineering methods to overexpress miR-378a-5p significantly enhanced the inhibitory effect of MSC-ApoVs on PANoptosis, while inhibiting miR-378a-5p weakened this protective effect. Conclusion This study reveals for the first time that APAP leads to hepatocyte PANoptosis. MSC-ApoVs block the formation of the PANoptosome complex through the "miR-378a-5p/NLRC5" axis, thereby alleviating APAP-induced acute liver failure. This provides a new strategy based on cell-free therapy for the treatment of APAP-ALF.</p>