机械生长因子对骨骼肌卫星细胞增殖及MGF mRNA表达的影响  

Extrinsic Mechano Growth Factor Induces Cell Proliferation and MGF mRNA Expression in Rat Skeletal Muscle Satellite Cells in Vitro

作　　者：史仍飞[1] 刘子青[2] 袁海平[1] 王顺利[1] 魏安奎[1] 张平[1] 

机构地区：[1]上海体育学院运动科学学院,上海200438 [2]上海市杨浦区中心医院康复医学科,上海200090

出　　处：《临床医学工程》2012年第1期18-21,共4页Clinical Medicine & Engineering

基　　金：国家自然科学基金资助项目(№30800542);上海市第二期重点学科建设项目资助(S30802)

摘　　要：目的骨骼肌卫星细胞是肌源性干细胞,该细胞的增殖受一系列生长因子的影响。而机械生长因子(mechano growth factor,MGF)是近年来发现的一种对力学信号敏感的自分泌局部生长因子,可以激活肌卫星细胞的增殖。本研究采用不同浓度的MGF干预体外培养的骨骼肌卫星细胞,探讨促进卫星细胞增殖的适宜浓度。方法用Ⅱ型胶原酶和胰蛋白酶两步酶消化法分离骨骼肌卫星细胞,待细胞同步化后,分别应用不同浓度的MGF(0ng/ml、25ng/ml、50ng/ml、100ng/ml和150ng/ml)干预原代培养的骨骼肌卫星细胞96小时,采用RT-PCR技术检测肌卫星细胞MGF mRNA的表达,用CCK-8检测卫星细胞的增殖情况,并检测细胞总蛋白含量。结果与0ng/ml MGF组相比,25ng/ml和50ng/ml组OD值在各时间点均显著性增加,其中25ng/ml组在24h至96h内OD值具有高度显著性(P<0.01),50ng/ml组在24h至72h内也有高度显著性的增加(P<0.01);25ng/ml和50ng/ml组MGF mRNA的表达量均显著高于对照组(P<0.05),其它组的表达与对照组无显著性差异。总蛋白含量也显示,与0ng/ml MGF组相比,25ng/ml和50ng/ml组均有显著性升高(P<0.05)。结论适宜浓度的MGF具有促进肌卫星细胞增殖的作用,其中浓度为25ng/ml和50ng/ml时具有较强的促增殖作用,但有一定的时间差异。Objective Mechano Growth Factor （MGF） is one kind of autocrine growth factor sensitive to mechanical stimulus. It plays a significant role between mechanical stimulation and gene expression by activating satellite cell proliferation. The purpose of the study is to investigate the effects of supplements of MGF on the proliferation of skeletal muscle satellite ceils. Methods Satellite cells were isolated from SD rat hind limb muscles with two-steps enzyme digestion. After cell synchronization, skeletal muscle satellite cells cultured for 96 hours, and different concentrations of MGF （0 ng/ml, 25 ng/ml, 50 ng/ml, 100 ng/ml and 150 ng/ml） were added to culture medium, respectively. The proliferation of satellite cells and the total protein content were detected. Total RNA was extracted from the cells samples and RT-PCR detection was used to evaluate mRNA MGF relative expression. Results Compared with 0 ng/ml MGF, OD values in 25 ng/m. 1 and 50 ng/ml group were significantly increased at each time point （P 〈0.05）, From 24 h to 96 h, OD value in 25 ng/ml group was highly significant （P〈0.01）, and OD value in 50 ng/ml group was also a highly significant increase at 24 h to 72 h （P〈0.01）; Compared with the control group, the expression of MGF mRNA in 25 ng/ml and 50 ng/ml groups were significantly increased （P 〈0.05）, but other treatment groups have no significant difference （P 〉0.05）; the total protein content in 25 ng/ml and 50 ng/ml group were significantly increased compared with 0 ng/ml group （P 〈0.05）. Conclusion The appropriate concentration of MGF has role in promoting muscle satellite cell proliferation, between 25 ng/ml and 50 ng/ml concentration have a strong effect on the proliferation, but difference in time course.

关 键 词：骨骼肌卫星细胞 机械生长因子 细胞原代培养 细胞增殖 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]