首页|基于HIF-VEGF-Notch通路探讨丁苯酞对氧糖剥夺/复氧人脑微血管内皮细胞的保护作用

基于HIF-VEGF-Notch通路探讨丁苯酞对氧糖剥夺/复氧人脑微血管内皮细胞的保护作用

扫码查看
目的 基于HIF-VEGF-Notch通路探讨丁苯酞对氧糖剥夺/复氧人脑微血管内皮细胞的保护作用.方法 制备氧糖剥夺/复氧人脑微血管内皮细胞模型,将细胞分为空白组、氧糖剥夺/复氧组、丁苯酞组和丁苯酞+sh-HIF-1α组.CCK-8及EdU染色检测H9c2细胞活力;流式细胞仪和AO/EB染色检测细胞凋亡;Matrigel体外成管试验检测血管生成能力;ELISA检测细胞中炎症因子含量;蛋白质印迹法检测细胞中H1F-1α、VEGF、Notch1蛋白表达.结果 和空白组相比,氧糖剥夺/复氧组细胞存活率(51.32±4.68)%、EdU阳性细胞率(12.08±0.75)%、小管形成数目(121.08±10.59)个明显降低,细胞凋亡率(14.92±1.06)%、细胞中IL-1β(30.46±2.51)pg/mL、IL-6(35.96±2.74)pg/mL、TNF-α(24.08±1.95)pg/mL 水平、HIF-1 α(0.61±0.08)、VEGF(0.72±0.08)、Notch 1(0.43±0.05)蛋白表达明显升高(P<0.01);和氧糖剥夺/复氧组相比,丁苯酞组细胞存活率(85.94±7.09)%、EdU 阳性细胞率(20.27±2.01)%、小管形成数目(204.15±16.71)个及细胞中 HIF-1α(1.15±0.12)、VEGF(0.96±0.10)、Notch1(0.87±0.10)蛋白表达均明显增加,细胞凋亡率(7.40±0.65)%、细胞中 IL-1 β(10.32±0.87)pg/mL、IL-6(12.81±1.03)pg/mL、TNF-α(10.31±0.89)pg/mL水平明显降低(P<0.0001);和丁苯酞组相比,丁苯酞+sh-HIF-1α组细胞存活率(54.18±5.06)%、EdU阳性细胞率(15.46±0.83)%、小管形成数目(129.26±11.64)个及细胞中 HIF-1 α(0.53±0.06)、VEGF(0.60±0.06)、Notch 1(0.36±0.04)蛋白表达明显降低,细胞凋亡率(12.87±1.01)%、细胞中 IL-1β(28.59±2.39)pg/mL、IL-6(32.87±2.31)pg/mL、TNF-α(22.01±1.87)pg/mL水平明显升高(P<0.01).结论 丁苯酞可促进氧糖剥夺/复氧人脑微血管内皮细胞血管生成,从而对损伤的细胞发挥保护作用,其作用机制可能和促进HIF-1α/VEGF/Notch1信号通路激活有关.
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.

ButylphthalideOxygen glucose deprivation/reoxygenationHuman brain microvascular en-dothelial cellsAngiogenesisHIF-1 α/VEGF/Notch1 signaling pathway

李娟、刘鑫、张宁、康永敬、宋娟、刘文红、陈永新

展开 >

黄骅市人民医院神经内一科,河北沧州 061100

丁苯酞 氧糖剥夺/复氧 人脑微血管内皮细胞 血管生成 HIF-1α/VEGF/Notch1信号通路

河北省沧州市重点研发计划指导项目

212302205

2024

解剖学研究
广东省解剖学会 中国解剖学会

解剖学研究

CSTPCD
影响因子:0.327
ISSN:1671-0770
年,卷(期):2024.46(5)