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.
QUAN Lu-Lu,LING Ming-Fa,LI Fan, et al. Regulatory Effect of VEGFB on Glucose Uptake During the Differentiation of Myoblast C2C12[J]. 农业生物技术学报, 2024, 32(6): 1362-1370.
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