People living long-term in areas with UV will cause premature photoaging. An abnormal reduction in autophagy is a key feature of photoaging, and p38MAPK has been regarded as a key regulator of autophagy. Isothiocyanate is one of the main active components of Moringa oleifera Lam. seeds. Studies have reported that MITC has anticancer, anti-inflammatory, cardiometabolic repair, nervous system protection, blood lipid regulation and diabetes prevention properties. However, the molecular mechanisms of MITC with protective effects against skin photoaging have not been studied thus far. In this study, we aimed to evaluate the antiphotoaging activity of MITC and to investigate the effect of p38MAPK-dependent autophagy in vivo and in vitro models of photoaging. In this research we found that MITC can reverse the intracellular ROS content and inhibit the activation of p38MAPK to improve the autophagy level, reduce the expression of MMPs, and finally protect against photoaging by UV. Our results will uncover the molecular mechanisms of MITC that play a role in the protective effects against skin photoaging, provide helpful information for developing MITC as an ant-photoaging plant material and improve the utilization of Moringa oleifera Lam. seeds.