Aim To investigate the effect of maternal perinatal high-salt diet on dimethylarginine dimethylamino-hydrolase 2 (DDAH2)/asymmetric dimethylarginine (ADMA)/endothelial nitric oxide synthase (eNOS)/nitric oxide (NO) pathway of the mesenteric artery in the male offspring rats. Methods The rats were divided into 2 groups:normal-salt diet (NSD) group and high-salt diet (HSD) group, and fed respectively with normal-salt diet (1%NaCl) and high-salt diet (8%NaCl) during perinatal period. After delivery, the male offspring rats were fed with the same diet for 16 weeks. Blood pressure, mesenteric artery endothelial-dependent diastolic function, NO content, eNOS activity and ADMA content in plasma and mesenteric artery, DDAH2 activity and protein expression of DDAH1 and DDAH2 in mesenteric artery were detected by various methods. Results The systolic blood pressure (SBP) in HSD group was significantly higher than that in NSD group (P<0.01). The endothelium-dependent tension of mesenteric artery in HSD group was lower than that in NSD group (P<0.01). After incubation with ADMA, the blood vessel tension was significantly decreased in NSD group, while no significant change was found in HSD group. Compared with the NSD group, NO content was decreased (P<0.05), eNOS activity was decreased (P<0.01), ADMA content was increased (P<0.05) in plasma in HSD group, and NO content was decreased (P<0.01), eNOS activity was decreased (P<0.01), ADMA content was increased (P<0.05) on mesenteric artery in HSD group. In HSD group, DDAH2 activity and protein expression were decreased (P<0.01), but DDAH1 protein expression was not changed significantly. In HSD group, correlation analysis of mesenteric arterial indexes showed that eNOS activity was positively correlated with NO content, ADMA content was negatively correlated with eNOS activity, DDAH2 activity and DDAH2 protein expression were negatively correlated with ADMA content. Conclusion The high-salt diet in the maternal perinatal period results in the increase of SBP and the endothelial-dependent diastolic dysfunction on mesenteric arteries in male offspring rats, which are related to the decrease of DDAH2 activity and the disorder of DDAH2/ADMA/eNOS/NO pathway in mesenteric arteries.