Vascular endothelial injury is a major cause of cardiovascular diseases. Conventional treatments have several limitations; we thus explored whether sophoridine could protect against tumor necrosis factor-alpha induced vascular endothelial injury. We evaluated the effects of sophoridine on cell survival and apoptosis in TNF-α-treated human umbilical vein endothelial cells and in mice. Western blotting and real-time quantitative reverse transcription polymerase chain reaction were used to estimate the expression levels of relevant proteins and their mRNAs. The in vivo effects of sophoridine on vascular endothelial function were explored by immunofluorescence staining. Sophoridine enhanced vascular endothelial cell survival and reduced apoptosis after tumor necrosis factor-α-induced injury, and modulated the expression levels of Src, Akt, PI3K, and ZO-1. Sophoridine protected against tumor necrosis factor-α induced vascular endothelial injury by regulating the Src/Akt/ PI3K pathway.