Objective To investigate the anti-oxidant effects of HXJN in brain microvascular endothelial cells (bEND.3) and the underlying mechanism. Methods HXJN-containing serum and HXJN extract solution were prepared and their effects on bEND.3 cell viability were measured by WST-1 assay. The protective effect of HXJN on H2O2-induced oxidative cell damage was also measured. Heme oxygenase (HO-1) mRNA level was detected by quantitative RT-PCR. The expression levels of HO-1, ph-Akt, ph-Nrf2 (nuclear factor erythroid 2, Nrf2) and Nrf2 nuclear translocation were detected by using Western Blot analysis. The involvement of Akt/Nrf2/HO-1 signaling pathway in the effect of HXJN were analyzed using specific pharmacological inhibitors. Results Treatment of the bEND.3 cells with HXJN-containing serum less than 10% for 24 h did not reduce the cell viability. Treatment of the cells with HXJN extract for 24 h increased the cell viability significantly. The cell viability was greatly decreased by a 3 h’s treatment of 200 μM or 300 μM H2O2, and HXJN showed a significant protective effect on the cell injury induced by 200 μM H2O2（P<0.01）. Furthermore, HXJN induced the expression of HO-1 and increased the phosphorylation and nuclear translocation of Nrf2. HXJN-induced HO-1 up-regulation was suppressed by LY294002, perifosine and ML385. The nuclear translocation of Nrf2 was inhibited by LY294002. Conclusion HXJN protects bEND.3 against oxidative stress through up-regulating the expression of HO-1. PI3K/Akt/Nrf2 signaling pathway may be involved in this effect.