Xiaoai Jiedu recipe promotes NLRP3 inflammasome activation by alleviating PINK1-mediated mitophagy in hepatocellular carcinoma

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Background The Xiaoai Jiedu recipe (XJR), a traditional Chinese medicinal formula, has been applied clinically for liver cancer management. XJR exhibits promising anti-tumor potential and may offer a novel strategy for oncological intervention. This study was designed to investigate the molecular mechanisms by which XJR exerts its pharmacological effects against hepatocellular carcinoma (HCC). Methods In vivo assays were conducted employing an HCC xenograft mouse model, and HepG2 cells were employed in vitro . ELISA, immunohistochemistry (IHC), immunofluorescence (IF), RT-PCR and western blot were performed to detect PTEN-induced putative kinase 1 (PINK1)-mediated mitophagy and NLRP3 (NOD-like receptor protein 3) inflammasome activation. The cellular mitochondrial reactive oxygen species (ROS) levels were tested with DCFH-DA and MitoSOX. Transmission electron microscopy (TEM) and JC-1 were utilized to observe the mitochondrial morphology and function, as well as mitochondrial membrane potential (MMP) in the process of XJR intervention for HCC. Results XJR prominently promoted pyroptosis by upregulating the related inflammatory cytokines levels in serum and HepG2 cells. The key targets include PINK1, parkin, p62, LC3B, TOMM20, COX IV, NLRP3, ASC, GSDMD, caspase1, IL-1β and IL-18 were observed. XJR disrupted PINK1-mediated mitophagy, which in turn increased mitochondrial ROS levels and promoted NLRP3 inflammasome assembly both in vitro and in vivo . Meanwhile, functional assays using pathway-specific inhibitors and siRNA-mediated knockdown further demonstrated that XJR suppresses mitophagy and enhances NLRP3 inflammasome activation via inhibition of the PINK1/parkin signaling pathway. Conclusions The preliminary results of the present study suggested that XJR may alleviate PINK1-mediated mitophagy and motivate NLRP3 signaling pathway to subsequently exert anti-tumor effects in HCC.