Protective effect of BNDF on vascular endothelial cells with H2 O2-induced oxidative injury / 中国病理生理杂志

ABSTRACT

[ABSTRACT]AIM:Tostudytheprotectiveeffectofbrain-derivedneurotrophicfactor(BDNF)onvascularendo-thelial cells with H 2 O2-induced oxidative injury .METHODS: Human umbilical vein endothelial cells ( HUVECs ) were cultured in vitro, and the oxidation injury model of HUVECs was established by treatment with H 2 O2 .The oxidatively in-jured HUVECs were cultured with different concentrations (1, 10 and 100μg/L) of BDNF.At the same time, the control group (no injury), PBS treatment after H2O2 injury group and TrkB inhibitor group (with 100 μg/L BDNF and 1∶1 000 TrkB inhibitor) were also set up.The viability of the HUVECs was detected by MTT assay .The levels of LDH, MDA, SOD and GSH were measured .The releases of NO , ET-1 and ICAM-1 were analyzed by ELISA .The changes of ROS pro-duction and cell apoptosis were evaluated by flow cytometry .The protein levels of TrkB , p-TrkB, cleaved caspase-3, Bcl-2 and Bax were determined by Western blot .RESULTS:Compared with uninjured control group , in H2 O2 oxidative injury plus PBS treatment group , the viability of the cells was decreased significantly , the LDH and MDA levels were increased significantly and the activities of SOD and GSH were decreased significantly .The NO secretion was decreased , and the ET-1 and ICAM-1 concentrations were increased significantly .The ROS content and apoptotic rate were increased significantly . The protein levels of cleaved caspase-3 and Bax were increased but Bcl-2 protein expression was decreased significantly . Compared with PBS treatment group , in H2 O2-injured HUVECs treated with different concentrations of BDNF , the cell via-bility was gradually increased , the LDH and MDA levels were decreased and the activities of SOD and GSH were increased gradually .The secretion of NO was increased but ET-1 and ICAM-1 were decreased gradually .The ROS content and apop-totic rate were decreased significantly .The TrkB and p-TrkB levels were significantly increased significantly , the protein expression of cleaved-caspase 3 and Bax was decreased gradually and the Bcl-2 protein expression increased gradually .The role of BDNF was inhibited by TrkB inhibitor .CONCLUSION:BDNF protects HUVECs from oxidative injury by binding with TrkB to activate the BDNF-TrkB signaling pathways .