In this study, we found that RIP1 inhibitor Nec‐1 inhibited the overproduction of ROS caused by osthole, whereas ROS inhibitor NAC also reduced the up‐regulated expression of RIP1, RIP3 and MLKL induced by osthole in U87 and C6 cells, which indicates that ROS plays an important role in the regulation of osthole‐mediated glioma cell necroptosis, and osthole may induce a positive feedback loop between ROS and RIP1. This evidence concerns the gene MLKL and glioma.