Hypoxia training (HT) can reduce body weight and improve fatty liver. However, the mechanism is not clear. A previous study indicated that HT-induced weight loss might be associated with the endocannabinoid system (ECS), which has also been reported recently to be involved in the persistent lipid mediators after weight loss. The present study investigated the effects of HT, a new prospective weight-loss method, on nutritionally obese mice and demonstrated that HT significantly reduced body weight, fat mass, transcriptional expression of liver endocannabinoid receptor 1 (CB1), biosynthetic enzyme diacylglycerol lipase alpha (DAGL alpha) and improved the transcriptional expression of degrading enzyme monoacylglycerol lipase (MAGL). Liver endocannabinoids 2-arachidonoylglycerol (2-AG) but not anandamide (AEA) was evidently decreased in response to HT. Simultaneously, HT significantly reduced liver index, serum alanine aminotransferase (ALT) and liver fat contents. Western blot showed decreased expression of liver CB1, sterol regulatory element-binding protein-1 (SREBP-1), peroxisome proliferator-activated receptor gamma (PPAR gamma) and increased expression of adipose triglyceride lipase (ATGL) and carnitine palmitoyltransferase-1 (CPT-1) levels after HT. However, intraperitoneal injection of CB1 receptor agonist WIN55212-2 offset the benefits by which HT reduced hepatic fat synthesis, with significant increased protein expression of SREBP-1 and PPAR gamma. Taken together, these findings reported the alleviation of obesity and hepatic steatosis through HT and provided a putative molecular mechanism by inhibiting the CB1-mediated fat synthesis. (C) 2020 Elsevier Inc. All rights reserved.