Abstract: Objective To investigate the effect of IH764-3 on the leukocyte-mediated hypoxia-reoxygention injury of human brain microvascular endothelial cell (HBMVEC). Methods MTT assay was used to detect the survival of HBMVEC; gelatin zymography was used to analyze the activity of MMPs. The level of reactive oxygen species （ROS） in leukocyte was determined via commercially available kit, and the indirect enzyme-linked immunosorbent assay （ELISA） was used to quantify the contents of TNF-α, IL-1α, IL-2 and INF-γ in leukocyte culture medium. Results Survival of HBMVEC was impaired by hypoxia-reoxygenation, and aggravated by supernatant of activated leukocytes, which was attenuated by IH764-3. Leukocytes produced high levels of MMP-9, ROS and cytokines (TNF-α, IL-1α, IL-2, IFN-γ) after suffered hypoxia-reoxygenation, which was inhibited by IH 764-3. Furthermore, IH 764-3 could effectively reverse hypoxia-reoxygenation injury of HBMVEC with supernatant of activated leukocytes. Conclusion IH764-3 can protect HBMVEC against leukocyte-involved hypoxia-reoxygenation injury by attenuating the activation of leukocytes and inhibiting the injury effects of leukocytes products.

Key words: IH764-3, Leukocytes, Brain, Endothelial Cells, Hypoxia-Reoxygenation injury