机械生长因子促骨骼肌卫星细胞增殖作用途径的初步研究

发布时间：2018-07-02 12:20

本文选题：机械生长因子 + 骨骼肌卫星细胞　；参考：《中国运动医学杂志》2014年11期

【摘要】：目的:研究机械生长因子促进骨骼肌卫星细胞增殖的量效关系及可能的作用途径。方法:选用4周龄雄性SD大鼠1只,无菌条件下取后肢肌肉,采用改进的两步酶消化法结合差速贴壁技术,分离及纯化骨骼肌卫星细胞。免疫细胞化学法鉴定骨骼肌卫星细胞。取第3代骨骼肌卫星细胞,随机分为对照组、DMEM对照组、MGF组(M组)、LY+MGF组(LM组)、PD+MGF组(PM组)和LY+PD+MGF组(LPM组),同步化后,相应组细胞换用含有PI3K特异性抑制剂LY294002(30μM)及MEK特异性抑制剂PD98059(50μM)的生长培养基预阻断12 h后,再用含有MGF(25 ng/ml)的生长培养基继续培养24 h后,用CCK-8比色法、BCA比色法观察不同抑制剂干预下SC的增殖及总蛋白含量。结果:1、PM组和LPM组OD值均显著低于M组(P0.01),而LM组OD值与M组无显著差异。2、LM组、PM组、LPM组SC总蛋白含量均显著低于M组。结论:MEK/Erk途径在MGF对卫星细胞促增殖方面有较重要作用。
[Abstract]:Aim: to study the dose-effect relationship and possible mechanism of mechanical growth factor (MGF) on proliferation of skeletal muscle satellite cells. Methods: a 4-week-old male Sprague-Dawley rat was used to isolate and purify skeletal muscle satellite cells under aseptic condition. Identification of skeletal muscle satellite cells by immunocytochemistry. The third generation skeletal muscle satellite cells were randomly divided into DMEM control group (MGF group), LY MGF group (LM group), PD MGF group (PM group) and LY PD MGF group (LPM group). The cells in the corresponding group were preblocked with PI3K-specific inhibitor LY294002 (30 渭 M) and MEK-specific inhibitor PD98059 (50 渭 M) for 12 h, then cultured on the growth medium containing MGF (25 ng/ml) for 24 h. The proliferation and total protein content of SC were observed by CCK-8 colorimetry and BCA colorimetry. Results the OD values of PM group and LPM group were significantly lower than those of M group (P0.01), but the OD value of LM group was not significantly different from that of M group. 2 the SC total protein content in LPM group was significantly lower than that in M group. Conclusion the MGF pathway plays an important role in promoting the proliferation of satellite cells.
【作者单位】：上海体育学院运动科学学院;
【基金】：上海市教委科研创新项目(05IZ02) 运动健身科技省部共建教育部重点实验室(上海体育学院)开放基金
【分类号】：R329.2

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