新藤黄酸通过PTEN-PI3K/AKT/VEGF/eNOS信号通路干预人脐静脉内皮细胞血管生成的研究

目的 探究PTEN-PI3K/Akt/VEGF/eNOS信号通路在新藤黄酸(gambogenic acid,GNA)抑制人脐静脉内皮细胞(human umbilical vein endothelial,HUVECs)迁移中的作用.方法 倒置显微镜下观察HUVECs形态变化;Boyden chamber实验以及细胞划痕...

Gambogenic Acid Intervenes Against Angiogenesis in Human Umbilical Vein Endothelial Cells via the PTEN-PI3K/AKT/VEGF/eNOS Signaling Pathway

Objective To investigate the role of the PTEN-PI3K/Akt/VEGF/eNOS signaling pathway in the inhibition of the migration of human umbilical vein endothelial cells (HUVECs) by gambogenic acid (GNA). Methods An inverted microscope was used to observe the morphological changes of HUVECs; Boyden chamber experiment and wound-healing assay were used to observe the effect of GNA on the migration ability of HUVECs; the nitrate reductase method was used to measure the level of nitric oxide (NO); Western blot was used to measure the effect of GNA and LY29400 pretreatment on the expression levels of phosphoinositide 3-kinase (PI3K), phosphorylated PI3K (p-PI3K), protein kinase B (AKT), phosphorylated AKT (p-AKT) in HUVECs; after HUVECs were transfected with PTEN-siRNA, RT-PCR was used to measure the mRNA expression of vascular endothelial growth factor (VEGF). Results GNA effectively inhibited the migration of HUVECs in a dose-dependent manner (P<0.05). GNA and the inhibitors LY294002 and L-NAME effectively inhibited the production of NO, and the GNA+LY294002 group and the GNA+L-NAME group had a significantly better inhibitory effect (P<0.05). Western blot showed that GNA upregulated the protein expression of PTEN and downregulated the protein expression of endothelial nitric oxide synthase, p-PI3K, and p-AKT (P<0.05). RT-PCR showed that after PTEN-siRNA transfection, there was a significant increase in the mRNA expression of VEGF (P<0.05). Conclusion GNA inhibits the migration of HUVECs and prevents angiogenesis in HUVECs via the PTEN-PI3K/Akt/VEGF/eNOS signaling pathway.