ABSTRACT
Objective To investigate the effect of IH764-3 on the leukocyte-mediated hypoxia-reoxygention injury of human brain microvascular endothelial cell (HBM VEC). Methods MTT assay was used to detect the survival of HBMVEC; gelatin zymography was used to check 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, which was aggravated by supernatant of activated leukocytes but was attenuated by IH764-3. Leukocytes produced high level of MMP-9, ROS and cytokines (TNF-α, IL-1α, IL-2, IFN-γ) after hypoxia-reoxygenation, the process was inhibited by IH 764-3. Furthermore, IH764-3 could effectively reverse hypoxia-reoxygenation injury of HBMVEC with supernatant of activated leukocytes. Conclusion IH764-3 can protect HBMVEC from leukocyte-involved hypoxia-reoxygenation injury by attenuating the activation of leukocytes and inhibiting the pathogenic effects of leukocytes products.
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 check 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-?,IL-1?,IL-2 and INF-? in leukocyte culture medium.Results Survival of HBMVEC was impaired by hypoxia-reoxygenation,which was aggravated by supernatant of activated leukocytes but was attenuated by IH764-3. Leukocytes produced high level of MMP-9,ROS and cytokines (TNF-?,IL-1?,IL-2,IFN-?) after hypoxia-reoxygenation,the process was inhibited by IH 764-3. Furthermore,IH764-3 could effectively reverse hypoxia-reoxygenation injury of HBMVEC with supernatant of activated leukocytes. Conclusion IH764-3 can protect HBMVEC from leukocyte-involved hypoxia-reoxygenation injury by attenuating the activation of leukocytes and inhibiting the pathogenic effects of leukocytes products.