Glut4突变调控脂肪重分布及肌纤维重塑的机制研究

doi:10.11843/j.issn.0366-6964.2021.09.009

Abstract

Abstract: The purpose of this study was to find out the molecular mechanism of skeletal muscle energy metabolism and muscle fiber transformation due to Glut4 gene mutation. Glut4^Q177L mutant mice were constructed by CRISPR/Cas9. The animals were divided into 2 groups including 22-week-old healthy wild male mice and Glut4^Q177L mutant male mice, 3 in each group, repeated 3 times. Subsequently, the phenotypic evaluation, glucose tolerance and insulin tolerance tests were performed. The expression of genes related to glucose transporters, lipid metabolism and muscle fiber type in gastrocnemius muscle of mice in the two groups were detected by qPCR; Western blot was applied to detect the content of AMPK and its phosphorylation level in gastrocnemius muscle. The results showed that the weight of subcutaneous fat and epididymal fat in mutant mice was significantly lower than that in the control group (P<0.05), and the area under the glucose tolerance curve of mutant mice significantly increased (P<0.01), indicated the glucose tolerance was impaired. The content of triglyceride in serum of mutant mice significantly decreased(P<0.05). The expression of several glucose transporters such as Glut4, Glut1 and Glut12 in gastrocnemius muscle of mutant mice was significantly higher than that in the control group (P<0.05). The limitation of glucose uptake resulted in a significant increase in the mRNA, protein and phosphorylation modification expression level of AMPK(P<0.05), which was the key gene regulating energy metabolism. At the same time, the expression of CD36 and ATGL, which promote fatty acid uptake and lipolysis, were significantly up-regulated (P<0.05). The expression of slow oxidation fiber type (type I) and fast oxidation fiber type (type IIa) related genes significantly increased(P<0.01), while the expression of fast glycolysis fiber type (type IIb) related gene significantly decreased(P<0.05). The results of this study showed that Glut4^Q177Lmutation reduced glucose uptake in skeletal muscle and fat tissue, resulting in compensating glucose uptake by up-regulating other glucose transporters. Besides, the insufficiency of energy could activate AMPK signaling pathway and secrete myokines to meet energy requirement by facilitating lipolysis. In addition, Glut4 mutation promoted the expression of slow oxidation fiber type related gene, rich in mitochondria, to improve energy efficiency. Glut4^Q177Lmice can not only provide an effective animal model of skeletal muscle insulin resistance, but also provide gene editing reference sites for animal breeding.

Key words: Glut4, insulin resistance, skeletal muscle, lipid metabolism, muscle fiber types

CLC Number:

S813.2

XIE Ning, ZHANG Kaiyi, RUAN Jinxue, TAO Cong, WU Tianwen, WANG Yanfang, YANG Shulin. Research of the Mechanism of Glut4 Mutation Regulating Fat Redistribution and Muscle Fiber Remodeling[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(9): 2464-2474.

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Research of the Mechanism of Glut4 Mutation Regulating Fat Redistribution and Muscle Fiber Remodeling

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