摘要
本文旨在探讨胰岛素对大鼠骨骼肌成肌细胞增殖的影响。分离和培养大鼠骨骼肌成肌细胞,用胰岛素作用于培养的骨骼肌成肌细胞,采用3H-TdR渗入法、BrdU检测法、MTT实验分析检测骨骼肌成肌细胞的增殖情况,用Western blot检测Akt和ERK的磷酸化水平,同时使用PI3K和MEK特异性抑制剂干预胰岛素对骨骼肌成肌细胞的增殖作用影响。结果显示,胰岛素能明显促进骨骼肌成肌细胞对3H-TdR的摄入,促进细胞的DNA合成和增殖,并且胰岛素促细胞增殖作用具有一定的量效关系,骨骼肌成肌细胞原代细胞和成肌细胞株L6、C2C12都显示该促增殖作用可被PI3K和MEK特异性抑制剂阻断,胰岛素还提高了骨骼肌成肌细胞Akt和ERK的磷酸化水平。本研究证实胰岛素可能通过PI3K/Akt和MEK/ERK信号通路促进骨骼肌成肌细胞增殖。
The present study was to explore the effects of insulin on proliferation of skeletal myoblast cells in rats. Separated and cul- tured primary skeletal myoblast cells from rats were treated by insulin. By means of the incorporation of 3H-TdR, BrdU assay and MTT assay, the proliferation of skeletal myoblast cells was detected. Western blot was used to check the phosphorylation of Akt and ERK of myoblast cells. The results showed that insulin significantly promoted the incorporation of 3H-TdR into cultured skeletal myo- blast cells in a dose-dependent manner. MTT assay and BrdU assay also showed insulin promoted the proliferation of skeletal myo- blast cells. The promotion of skeletal myoblast cells proliferation by insulin was inhibited by PI3K inhibitor wortmannin or MEK in- hibitor U0126, and the same phenomenon was shown in L6 and C2C12 cells. Also, insulin increased the phosphorylation of Akt and ERK in myoblast cells. These results suggest that insulin may promote proliferation of skeletal myoblast cells through PI3K/Akt and MEK/ERK pathways.
出处
《生理学报》
CAS
CSCD
北大核心
2013年第1期19-25,共7页
Acta Physiologica Sinica
基金
supported by the Key Project of Science and Technology from Education Department of Jiangxi Province
China(No.GJJ11694)
the National Natural Science Foundation of China(No.81000075)
