Pyroptosis Pathway Activation and NLRP3 Inflammasome Biochemistry in Patients with Non-Alcoholic Fatty Liver Disease in Kano

Pyroptosis, a form of inflammatory programmed cell death mediated by the NLRP3 inflammasome and gasdermin D, is an emerging mechanism driving hepatocyte injury in non-alcoholic fatty liver disease, and its biochemical characterisation in West African populations is entirely unexplored. This study investigates NLRP3 inflammasome activation, caspase-1 activity, gasdermin D cleavage, and pyroptotic cytokine release in liver biopsy tissue and peripheral blood mononuclear cells of NAFLD patients in Kano, North West Nigeria. A case-control design was employed, recruiting 60 biopsy-confirmed NAFLD patients including 30 with simple steatosis and 30 with NASH, and 40 healthy controls. NLRP3 protein expression and ASC speck formation were assessed by immunohistochemistry and Western blotting. Caspase-1 and caspase-4/5 activities were measured by fluorometric assay. Plasma interleukin-1 beta, interleukin-18, and gasdermin D N-terminal fragment were quantified by ELISA. Pyroptotic activity was correlated with histological inflammation score (NAFLD Activity Score) and fibrosis stage. Transcriptomic analysis by RNA sequencing of PBMC was integrated to construct a pyroptosis gene regulatory network. This study generates original mechanistic biochemistry on pyroptosis in NAFLD within a West African population context and identifies novel therapeutic targets. Keywords: pyroptosis, NLRP3 inflammasome, NAFLD, gasdermin D, Kano.