VEGFB对成肌细胞C2C12分化过程中葡萄糖摄取的调控作用
Regulatory Effect of VEGFB on Glucose Uptake During the Differentiation of Myoblast C2C12
全露露 1凌明发 1李凡 1刘金好 1江青艳 1王松波1
作者信息
- 1. 华南农业大学动物科学学院/广东省动物营养调控重点实验室,广州 510642
- 折叠
摘要
血管内皮生长因子B(vascular endothelial growth factor B,VEGFB)是骨骼肌摄取葡萄糖的重要调控因子,VEGFB敲除可促进糖尿病小鼠(Mus musculus)骨骼肌组织对葡萄糖的摄取,改善糖尿病小鼠的葡萄糖不耐受和胰岛素抵抗.但是,目前VEGFB对骨骼肌细胞葡萄糖摄取的具体调控作用尚未阐明.为探究VEGFB对骨骼肌细胞葡萄糖摄取的影响,本研究以小鼠成肌细胞C2C12为模型,利用qPCR技术检测C2C12分化过程中VEGF信号相关基因和葡萄糖转运载体4(glucose transporter 4,GLUT4)的表达,采用葡萄糖氧化酶法和2-NBDG(2-(N-(7-nitrobenz-2-oxa-13-diazol-4-yl)amino)-2-deoxy-D-glucose)法检测VEGFB、VEGF受体(VEGF receptor,VEGFR)抑制剂Axitinib和磷酸肌醇3激酶(phosphatidylinositol 3 kinase,PI3K)抑制剂Wortmanin对C2C12分化过程中葡萄糖消耗量和摄取量的影响,采用Western blot检测GLUT4在细胞膜上及胞浆内的表达以及PI3K和丝氨酸/苏氨酸蛋白激酶(serine/threonine kinase proteins,AKT)的激活情况.结果显示,与分化前相比,C2C12分化后VEGFB(P＜0.01)、VEGFR2(P＜0.01)和GLUT4(P＜0.05)的基因表达显著增加;外源添加VEGFB显著增加C2C12在分化过程中对葡萄糖的摄取(P＜0.05),显著促进GLUT4在细胞膜上的表达(P＜0.05),同时激活PI3K/AKT通路;而VEGF受体抑制剂Axitinib可逆转VEGFB对葡萄糖摄取、GLUT4膜转位及PI3K/AKT激活的促进作用;此外,PI3K抑制剂Wotmanin处理也可逆转VEGFB对葡萄糖摄取及GLUT4细胞膜表达的促进作用.上述研究结果表明,VEGFB可能通过VEGFR-PI3K/AKT信号通路促进GLUT4向C2C12细胞膜转位,进而促进C2C12在分化过程中对葡萄糖的摄取.本研究揭示了VEGFB对成肌细胞C2C12分化过程中葡萄糖摄取的调控作用,为骨骼肌葡萄糖代谢调控提供了新的科学认识,也为深入研究骨骼肌葡萄糖代谢的调控机制提供基础依据.
Abstract
Vascular endothelial growth factor B(VEGFB)is an important regulator of glucose uptake in skeletal muscle.Knockout of VEGFB can promote glucose uptake in skeletal muscle tissue of diabetes mice(Mus musculus),and improve glucose intolerance and insulin resistance in diabetes mice.However,there is no research to clarify the specific regulation of VEGFB on glucose uptake of skeletal muscle cell.In order to explore the regulatory effect of VEGFB on glucose uptake by skeletal muscle cell,this study used mouse myoblast C2C12 as a model,and used qPCR to detect the gene expression of VEGF signaling related genes and glucose transporter 4 gene(GLUT4)during the differentiation process of C2C12.The effects of VEGFB,VEGF receptor(VEGFR)inhibitor Axitinib and PI3K inhibitor Wortmanin on glucose consumption and uptake during C2C12 differentiation were detected by glucose oxidase method and 2-(N-(7-nitrobenz-2-oxa-13-diazol-4-yl)amino)-2-deoxy-D-glucose(2-NBDG)method.Western blot was used to detect the expression of GLUT4 on the cell membrane and in the cytoplasm and the activation of PI3K/AKT.The results of qPCR showed that the gene expression of VEGFB(P＜0.01),VEGFR2(P＜0.01)and GLUT4(P＜0.05)increased significantly after differentiation of C2C12 compared with that before differentiation.Exogenous addition of VEGFB significantly increased the glucose uptake of C2C12 during differentiation(P＜0.05),and significantly promoted the expression of GLUT4 on the cell membrane(P＜0.05),and activated the PI3K/AKT pathway.But the VEGF receptor inhibitor Axitinib could reverse the promotion of VEGFB on glucose uptake,GLUT4 membrane localization and PI3K/AKT activation.In addition,PI3K inhibitor Wotmanin treatment could also reverse the effect of VEGFB on glucose uptake and GLUT4 cell membrane expression.The results of this study indicate that VEGFB could promote GLUT4 translocation to C2C12 cell membranes through the VEGFR-PI3K/AKT signaling pathway,thereby promoting glucose uptake by C2C12 during differentiation.This study reveals the regulatory effect of VEGFB on glucose uptake during the differentiation of myoblast C2C12,providing a new scientific understanding of the regulation of skeletal muscle glucose metabolism,and a basic reference for in-depth research on the regulation mechanism of skeletal muscle glucose metabolism.
关键词
血管内皮生长因子B(VEGFB)/C2C12细胞系/葡萄糖摄取/PI3K/AKTKey words
Vascular endothelial growth factor B(VEGFB)/C2C12 cell line/Glucose uptake/PI3K/AKT引用本文复制引用
基金项目
国家自然科学基金(31972636)
国家自然科学基金(32172682)
出版年
2024
国家科技期刊平台
NSTL
万方数据