The twinning and hydrogen embrittlement (HE) behavior of twinning-induced plasticity (TWIP) steels with various gradient twin structures were investigated by slow strain rate tensile testing. The results revealed that the gradient design can be very beneficial for alleviating the strength-ductility trade-off dilemma in TWIP steels, especially under hydrogen environment. Among the different structures after cathodic hydrogen charging, the most outstanding was the inverse gradient-like structure material, which comprehensively utilized fine recrystallized grains at sample’s edge region to effectively lower HE (EIloss: 4.01% and UTSloss: 1.61%) and gradient-design of the central coarse grain to improve the comprehensive mechanical properties.