LiCl激活经典Wnt信号通路诱导猪骨骼肌卫星细胞向慢肌分化

Wnt/β-catenin信号通路参与调控细胞的增殖及分化,决定细胞的命运。LiCl通过抑制GSK-3β(glycogen synthetase kinase-3β)稳定细胞内β-catenin的表达。为研究Wnt/β-catenin信号通路在哺乳动物骨骼肌卫星细胞成肌分化过程中的作用。以原代培养猪骨骼肌卫星细胞为实验材料,将其培养在含有1%CEE和20%胎牛血清的GMEM培养液中,再用15 mmol/L LiCl诱导分化,并采用免疫荧光染色和Western blotting等方法检测经典Wnt信号通路中相关因子及成肌分化过程中细胞的蛋白表达。结果表明:LiCl处理猪骨骼肌卫星细胞后促进分化,p-GSK-3β、β-catenin、myosin及慢肌蛋白增加,p-β-catenin、GSK-3β及快肌蛋白减少,并且核内源性β-catenin增多。结果提示,激活Wnt/β-catenin信号通路促进骨骼肌卫星细胞成肌分化,并且诱导其向慢肌分化。

Activation Canonical Wnt Signaling by LiCl Induces Porcine Skeletal Muscle Satellite Cells Differentiation into Slow Muscle

Wnt/β-catenin signaling involved in regulating cell proliferation and differentiation to determine cell fate.Lithium chloride(LiCl) is known to activate canonical Wnt signaling by inhibiting glycogen synthetase kinase-3β and consequently stabilizing free cytosolic β-catenin.To understand the role of the Wnt/β-catenin pathway in the regulation of porcine skeletal muscle satellite differentiation,to make the skeletal muscle satellite cells for the new sources of the diseases treatment.We cultured porcine skeletal muscle satellite cells in GMEM with 1% CEE,20% FBS,and supplied 15 mmol/L LiCl to induce myoblast differentiation.Then we detected the protein expression of Wnt/β-catenin related genes and myoblast differentiation marker genes by Western blotting and immunofluorescence.The results showed that the cells protein of p-GSK-3β,β-catenin,myosin and MYH7 were increased,p-β-catenin GSK-3β and MY-32 were decreased after LiCl treatment.Additionally,the nuclear staining of endogenousβ-catenin was also significantly increased.These result demonstrated that Wnt/β-catenin is a significant pathway that regulates myogenic differentiation,and induced porcine skeletal muscle satellite cells differentiation to slow muscle.