Exploring the protective effect of butylphthalide on oxygen glucose deprivation/reoxygenation human brain microvascular endothelial cells based on the HIF-VEGF Notch pathway
Objective To investigate the protective effect of butylphthalide on oxygen glucose deprivation/reoxygenation human brain microvascular endothelial cells based on the HIF-VEGF Notch pathway.Method A hu-man brain microvascular endothelial cell model was prepared by oxygen glucose deprivation/reoxygenation,and the cells were divided into blank group,oxygen glucose deprivation/reoxygenation group,butylphthalide group,and bu-tylphthalide+sh-HIF-1 group α Group.CCK-8 and EdU staining were used to detect the activity of H9c2 cells;Flow cytometry and AO/EB staining were used to detect cell apoptosis;Matrigel in vitro tube forming test to detect angio-genesis ability;ELISA detection of inflammatory factor content in cells;Protein blotting method for detecting HIF-1 in cells α,VEGF and Notch 1 protein expression.Results Compared with the control group,the cell survival rate(51.32%±4.68%),EdU-positive cell rate(12.08%±0.75%),number of tubule formation(121.08±10.59),cell apoptosis rate(14.92%±1.06%),levels of IL-1 β(30.46 pg/mL±2.51 pg/mL),IL-6(35.96 pg/mL±2.74 pg/mL),TNF-α(24.08 pg/mL±1.95 pg/mL),protein expression of HIF-1α(0.61±0.08),VEGF(0.72±0.08),and Notch 1(0.43±0.05)were significantly decreased in the oxygen-glucose deprivation/reoxygenation(OGD/R)group(P<0.01).Compared with the OGD/R group,the cell survival rate(85.94±7.09)%,EdU-positive cell rate(20.27%±2.01%),number of tubule formation(204.15±16.71),protein expression of HIF-1α(1.15±0.12),VEGF(0.96±0.10),and Notch1(0.87±0.10)were significantly increased,while cell apoptosis rate(7.40±0.65)%,levels of IL-1β(10.32 pg/mL±0.87 pg/mL),IL-6(12.81 pg/mL±1.03 pg/mL),TNF-α(10.31 pg/mL±0.89 pg/mL)were signifi-cantly decreased in the butein group(P<0.0001).Compared with the butein group,the cell survival rate(54.18%±5.06%),EdU-positive cell rate(15.46%±0.83%),number of tubule formation(129.26±11.64),protein expres-sion of HIF-1α(0.53±0.06),VEGF(0.60±0.06),and Notch1(0.36±0.04)were significantly decreased,while cell apoptosis rate(12.87±1.01)%,levels of IL-1β(28.59 pg/mL±2.39 pg/mL),IL-6(32.87 pg/mL±2.31 pg/mL),TNF-α(22.01 pg/mL±1.87 pg/mL)were significantly increased in the butein+sh-HIF-1α group(P<0.01).Conclusion Butylphthalide can promote angiogenesis in oxygen glucose deprivation/reoxygenation human brain microvascular en-dothelial cells,thereby exerting a protective effect on damaged cells.Its mechanism of action may be related to the promotion of HIF-1 α/VEGF/Notch1 signaling pathway activation is involved.