Objectives: To investigate the role and mechanism of fibroblast growth factor (FGF) 19 in inflammation-induced injury of vascular endothelial cells caused by high glucose (HG).
Methods: Human umbilical vein endothelial cells (HUVECs) were randomly divided into four groups: control, HG, FGF19, and HG+FGF19 (n=3 each). The effect of different concentrations of glucose and/or FGF19 on HUVEC viability was assessed using the CCK8 assay. Flow cytometry was utilized to examine the impact of FGF19 on HUVEC apoptosis. Levels of interleukin-6 (IL-6), inducible nitric oxide synthase (iNOS), total superoxide dismutase (T-SOD), and malondialdehyde (MDA) were measured by ELISA. Real-time quantitative PCR and Western blotting were used to determine the mRNA and protein expression levels of vascular endothelial growth factor (VEGF), nuclear factor erythroid 2 related factor 2 (Nrf2), and heme oxygenase-1 (HO-1). Cells were further divided into control, siRNA-Nrf2 (siNrf2), HG, HG+FGF19, HG+FGF19+negative control, and HG+FGF19+siNrf2 groups (n=3 each) to observe the effect of FGF19 on oxidative stress injury in HUVECs induced by high glucose after silencing the Nrf2 gene.
Results: Compared to the control group, the HG group exhibited increased apoptosis rate, increased IL-6, iNOS and MDA levels, and increased VEGF mRNA and protein expression, along with decreased T-SOD activity and decreased mRNA and protein expression of Nrf2 and HO-1 (P<0.05). Compared to the HG group, the HG+FGF19 group showed reduced apoptosis rate, decreased IL-6, iNOS and MDA levels, and decreased VEGF mRNA and protein expression, with increased T-SOD activity and increased Nrf2 and HO-1 mRNA and protein expression (P<0.05). Compared to the HG+FGF19+negative control group, the HG+FGF19+siNrf2 group had decreased T-SOD activity and increased MDA levels (P<0.05).
Conclusions: FGF19 can alleviate inflammation-induced injury in vascular endothelial cells caused by HG, potentially through the Nrf2/HO-1 signaling pathway.
目的: 探讨成纤维细胞生长因子（fibroblast growth factor, FGF）19在高糖（high glucose, HG）导致的血管内皮细胞炎症损伤中的作用及其机制。方法: 将人脐静脉内皮细胞（human umbilical vein endothelial cell, HUVEC）随机分为对照组、HG组、FGF19组、HG+FGF19组（n=3）。运用CCK8法检测不同浓度葡萄糖和/或FGF19对HUVEC细胞活力的影响，流式细胞术检测FGF19对HUVEC细胞凋亡的影响，ELISA法测定白细胞介素-6（interleukin 6, IL-6）、诱导型一氧化氮合酶（inducible nitric oxide synthase, iNOS）、总超氧化物歧化酶（total superoxide dismutase, T-SOD）、丙二醛（malondialdehyde, MDA）水平，实时荧光定量PCR及Western blot法检测血管内皮生长因子（vascular endothelial growth factor, VEGF）、红系衍生的核因子2相关因子2（nuclear factor erythroid 2 related factor 2, Nrf2）、血红素加氧酶-1（heme oxygenase-1, HO-1）mRNA及蛋白表达水平；另取细胞分为对照组、siRNA-Nrf2（siNrf2）组、HG组、HG+FGF19组、HG+FGF19+阴性对照组、HG+FGF19+siNrf2组（n=3），观察沉默Nrf2基因后FGF19对HG诱导的HUVEC氧化应激损伤的影响。结果: 与对照组比较，HG组细胞凋亡率和IL-6、iNOS、MDA含量以及VEGF mRNA及蛋白表达升高（P<0.05），T-SOD活力以及Nrf2、HO-1 mRNA及蛋白表达降低（P<0.05）；与HG组比较，HG+FGF19组细胞凋亡率和IL-6、iNOS、MDA含量以及VEGF mRNA及蛋白表达降低（P<0.05），T-SOD活力以及Nrf2、HO-1 mRNA及蛋白表达水平升高（P<0.05）。与HG+FGF19+阴性对照组相比，HG+FGF19+siNrf2组T-SOD活力下降，MDA含量升高（P<0.05）。结论: FGF19可减轻HG导致的血管内皮细胞炎症损伤，其机制可能与Nrf2/HO-1信号通路有关。.
Keywords: Fibroblast growth factor19; Human umbilical vein endothelial cell; Inflammation; Oxidative stress; Type 1 diabetes mellitus; Vascular endothelial dysfunction.