We found that long term HFD significantly facilitated the protein expressions of both ER stress markers and pyroptosis markers including GRP78,CHOP(ER stress markers), NLRP3, GSDMD/−N, Caspase-1, p20, and IL-1 beta (pyroptosis markers), suggesting that ER stress and pyroptosis are important pathological process of NAFLD. This evidence concerns the gene GSDMD and metabolic dysfunction-associated steatotic liver disease.