Ide基因通过AKT调控成肌细胞增殖和分化的研究

doi:10.11843/j.issn.0366-6964.2020.08.002

Abstract

Abstract: This study was performed to investigate the expression of insulin-degrading enzyme (Ide) in different tissues of pig as well as its role in C2C12 myoblast proliferation and differentiation. RT-PCR and Western blotting were used to detect the expression of Ide in different tissues of 8-month old male mini-pigs. siRNA technology was used to knock down the expression of Ide in mouse C2C12 cell line to explore the role of Ide in myoblast proliferation,apoptosis and differentiation. Two groups containing Ide-siRNA treatment group and NC-siRNA (negative control) group were set up. There were 3 replicates in each group. The results showed that Ide was widely expressed in brain, quadriceps, biceps femoris, longest dorsal muscle, heart, liver, spleen, lung, kidney, and testis of pigs, among which its expression was relatively higher in quadriceps, kidney, and testis. After Ide-siRNA transfection into myoblasts for 48 hours, Ide expression was extremely significantly decreased(P<0.001). CCK-8 analysis showed that myoblast proliferation was promoted after Ide interference. Accordingly, the expressions of cell cycle-related genes were significantly increased. Meanwhile, the down-regulation of Ide by siRNA interference did not lead to myoblast apoptosis. Myoblasts induced to differentiation by 2% horse serum were transfected with Ide-siRNA, which resulted in the significant decrease in expression of differentiation marker genes Myog and Myhc at Day 2 and Day 5 compared with control group. This implied that the differentiation of myoblasts was inhibited after Ide interference. We further found that the expressions of Akt2 and phosphorylated AKT (P-AKT) were decreased at Day 5 of differentiation. In summary, Ide was widely expressed in different tissues of pigs, with higher expression in muscle, kidney and testis. Ide is involved in negatively regulating the proliferation of myoblasts, and interfering Ide can inhibit the differentiation of myoblasts through Akt2/P-Akt/Myog pathway.

Key words: insulin-degrading enzyme(Ide), pig, myoblast, proliferation and differentiation, AKT

CLC Number:

S813.3

GUO Jiankang, FAN Ziyao, NIU Pengxia, LIU Zhiguo, MU Yulian, ZHANG Mingrui, LI Kui, WANG Bingyuan. The Study of Ide Gene Regulating Myoblast Proliferation and Differentiation through AKT[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(8): 1784-1794.

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The Study of Ide Gene Regulating Myoblast Proliferation and Differentiation through AKT

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