<p>Metabolic dysfunction-associated steatotic liver disease (MASLD) has become a major global public health problem, and its occurrence is associated with adverse environmental exposures during development. In this study, we investigated the impact of the use of prednisone (a synthetic glucocorticoid drug) during pregnancy on susceptibility to MASLD in offspring and explored its potential therapeutic targets. Pregnant rodents were administered clinically equivalent doses of prednisone daily by oral gavage during gestation days (GDs) 0–20 in rats and GD0–18 mice, and their offspring were fed a high-fat diet from postnatal weeks 8–12. The results showed that prenatal prednisone exposure (PPE) led to reduced hepatic glucose uptake and fatty acid oxidation in offspring rats prenatally and postnatally and that the offspring developed more severe MASLD when fed a high-fat diet, with males exhibiting greater severity than females. Consistent findings were observed in PPE adult offspring mice. RNA-seq and experimental results revealed that hepatic Serpina3c expression was consistently reduced in PPE offspring before and after birth, which led to an increase in chymase-Ang II production and subsequent activation of its receptor AT1R, leading to MASLD susceptibility. In vivo and in vitro studies revealed that the programming of low Serpina3c expression was associated with reduced H3K27ac levels in the gene promoter region of Serpina3c caused by the activation of GR-HDAC3 signaling by the active metabolite prednisolone. Finally, postnatal high expression of hepatic Serpina3c reversed the activation of the chymase-Ang II-AT1R pathway and significantly ameliorated hepatic glucose and lipid metabolic dysfunction and MASLD susceptibility in PPE offspring. In summary, this study reveals MASLD susceptibility in offspring induced by PPE and identifies Serpina3c as a target for the prevention and treatment of MASLD susceptibility.</p>