Activation of the NLRP3 inflammasome by CCl₄ exacerbates hepatopathogenic diet-induced experimental NASH

Administration of carbon tetrachloride (CCl₄), along with an hepatopathogenic diet, is widely employed as a chemical inducer to replicate human nonalcoholic steatohepatitis (NASH) in rodents; however, the role of the nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 (NLRP3) inflammasome in this model remains unclear. We aimed to determine the relevance of NLRP3 inflammasome activation in the development of NASH induced by CCl₄ along with an hepatopathogenic diet in male Wistar rats.

Animals were fed either a high fat, sucrose, and cholesterol diet (HFSCD) or a HFSCD plus intraperitoneal injections of low doses of CCl₄ (400 mg/kg) once a week for 15 weeks. Liver steatosis, inflammation, fibrosis, and NLRP3 inflammasome activation were evaluated using biochemical, histological, ultrastructural, and immunofluorescence analyses, western blotting, and immunohistochemistry.

Our experimental model reproduced several aspects of the human NASH pathophysiology. NLRP3 inflammasome activation was induced by the combined effect of HFSCD plus CCl₄ and significantly increased levels of both proinflammatory and profibrogenic cytokines and collagen deposition in the liver; thus, NASH severity was higher in the HFSCD+CCl₄ group than that in the HFSCD group, to which CCl₄ was not administered. Hepatic stellate cells, the most profibrogenic cells, were activated by HFSCD plus CCl₄, as indicated by elevated levels of α-smooth muscle actin. Thus, activation of the NLRP3 inflammasome, triggered by low doses of CCl₄, exacerbates the severity of NASH.

Our results indicate that NLRP3 inflammasome activation plays a key role and may be an important therapeutic target for NASH treatment.