The development of new renewable bio-based resins is an important means to realize the substitution of petroleum-based related products and green and sustainable development. In this paper, epoxy soybean oil-based acrylic resin (AESO) was prepared by one-pot method using maleic anhydride (MA), methyl tetrahydrophthalic anhydride (MTHPA), hydroxyethyl acrylate (HEA) and epoxy soybean oil (ESO) as raw materials. A certain amount of isocyanate or HEA semi-sealed isocyanate was added to the resin to obtain an epoxy soybean oil-based acrylate-polyurethane resin with a bio-base content of more than 30%. The effects of different isocyanate types and amounts on photocuring activity, thermal stability and tensile properties of resin were investigated. Through the dynamic thermo-mechanical, mechanical properties and rheological viscosity tests, the results show that the introduction of HEA structure can increase the glass transition temperature to 65.3 ℃, the energy storage modulus to 1417.9 MPa. the tensile strength is up to 27.4 MPa and the fracture elongation is 67.2%. The viscosity is reduced to 43.4 Pa·s. At the same time, the gel-found rate of the above two resins is more than 86%, both of which have alkali soluble, excellent solvent resistance and high gloss. This study provided a new idea for the development of epoxy soybean oil-based acrylate-polyurethane resin, which is helpful to broaden its application in the field of bio-based light curing coating.