Here, we demonstrated that gut microbiota exerted protective effects against HFD-induced obesity and improved glucose homeostasis associated with several mechanisms involving gut microbial metabolites of dietary PUFAs: (1) gut microbiota converted LA to HYA, reducing the adipose inflammation otherwise induced by excessive dietary LA via the AA cascade; (2) HYA activated GPR40 and GPR120 and promoted GLP-1 secretion; and (3) HYA suppressed lipid absorption by promoting intestinal peristalsis via EP3. Here, PTGER3 is linked to obesity due to melanocortin 4 receptor deficiency.