The severity of non-alcoholic steatohepatitis (NASH) is determined by environmental and genetic factors, the latter of which are incompletely characterized. Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1) is a 130 kDa transmembrane glycoprotein expressed on blood and vascular cells. In the present study, we provide data for the novel finding that genetic deficiency of PECAM-1 potentiates the development and progression of NASH. We found that the rate of development and severity of diet-induced NASH are markedly enhanced in PECAM-1-deficient (KO) relative to wild-type (WT) mice, as measured by histological and biochemical evaluation. Livers from KO mice exhibited typical histological features of NASH, including macrovesicular fat accumulation, hepatocyte injury with infiltration of inflammatory cells, fibrosis and heightened oxidative stress. Alanine aminotransferase, a marker for liver injury, was also significantly higher in KO compared to WT mice. Consistent with a role for PECAM-1 as a suppressor of pro-inflammatory cytokines, plasma levels of inflammatory cytokines, including TNF- and MCP-1, were also significantly higher in KO compared to WT mice. These findings are the first to show that the PECAM-1-deficient mouse develops progressive NAFLD, supporting a role for PECAM-1 as a negative regulator of NAFLD progression. Future examination of recently-identified PECAM-1 allelic isoforms in humans as potential risk factors for developing NASH may be warranted.