Update information. 10-hydroxydec-2-enoic acid (10-HDA), a natural agent from royal jelly, could fight against cell and tissue damage, yet the underlying mechanisms are still unexplored. Here, we found the scavenging rate of hydroxyl free radical (•OH) by 2% (g/mL) 10-HDA was more than 20%, which was achieved by a multiple-step oxidization of the −OH group and C=C bond in 10-HDA. Moreover, 10-HDA could enhance the viability of vascular smooth muscle cells (VSMCs) damaged by •OH, indicating its role in reducing the •OH-induced cytotoxicity. The 195 proteins decreased by •OH challenge in VSMCs were up-regulated by 10-HDA rescue, which mainly elevated the activities of protein and energy metabolisms within nucleus and cytoplasm. These evidences indicate that 10-HDA plays key roles in promoting cell survival through antagonizing •OH-induced injury in VSMCs. Our results provide the first solid evidence that 10-HDA scavenging •OH could be a potential pharmacological application in keeping vascular health.