Synthesis of highly-efficiency electrocatalysts with controllable morphology, enriched active sites, and low cost is of great significance for urea oxidation reaction (UOR) applications. In this work, novel two-dimensional (2D) nickel-based metal-organic framework (Ni-MOF) nanosheets comprising Ni2+ and an organic ligand of 4-Dimethylaminopyridine (Ni-DMAP-t, where t is the synthesis time) are firstly synthesized by a facile one-pot solvothermal method. By thickness regulation of Ni-DMAP-t nanosheets, the self-supported Ni-DMAP-2 with the smallest thickness on Ni foam (Ni-DMAP-2/NF) exhibits high electrocatalytic activity toward UOR with a low potential of 1.45 V at a current density of 100 mA cm−2. Moreover, by further coupling with the cathodic hydrogen evolution reaction, the urea-assisted electrolytic hydrogen production system using Ni-DMAP-2/NF as anode displays a dramatic voltage reduction by 290 mV at a current density of 50 mA cm−2 as compared with that of conventional water electrolysis using the same electrodes. Such a superior UOR electrocatalytic activity of Ni-DMAP-2/NF is ascribed to the abundant Ni active sites exposed on the surface of Ni-DMAP-2, as well as the accelerated electron/ion transfer properties endowed by its ultrathin 2D nanosheets structure. This work offers some new ideas into the design and development of ultrathin 2D Ni-based MOF electrocatalysts for urea-related energy storage devices. Display omitted •Ni-MOFs comprising Ni2+ and 4-Dimethylaminopyridine are firstly synthesized.•The Ni-DMAP-2 with the smallest thickness shows remarkable UOR activity.•A urea-assisted H2 production with reduced cell voltage is constructed.