机械牵拉对大鼠骨骼肌卫星细胞的增殖作用及其途径的研究  

The Effects and Mechanisms of Mechanical Stretch on the Proliferation of Satellite Cells in Rat Skeletal Muscle

作　　者：杨璐 陆耀飞[1] YANG Lu;LU Yaofei(Shanghai University of Sport,Department of Kinesiology,Shanghai 200438,China)

机构地区：[1]上海体育学院运动科学学院

出　　处：《体育科学》2019年第10期83-90,共8页China Sport Science

摘　　要：目的:探讨不同的牵拉模式对大鼠骨骼肌卫星细胞增殖的作用及其可能的作用途径。方法:用两步酶消化法和差速贴壁离心技术分离和提纯卫星细胞。取第3代卫星细胞,利用Flexcell细胞牵拉系统以频率为1 Hz,分别施加5%和15%的拉伸度,各牵拉4 h和6 h。CCK-8检测牵拉结束24 h后卫星细胞的增殖情况,WB检测牵拉结束即刻和牵拉结束24 h后Akt、P38、ERK和JNK的活性以及牵拉结束24 h后的Myf5、MyoD的表达。结果:1)与对照组(不牵拉,下同)相比,15%拉伸度,牵拉4 h和6 h骨骼肌卫星细胞增殖升高(P<0.01),5%拉伸度,牵拉4 h和6 h未出现显著增殖(P>0.05);2)15%拉伸度,牵拉6 h的Myf5蛋白表达高于对照组(P<0.01),牵拉4 h和6 h的MyoD的表达高于对照组(P<0.01),5%拉伸度,牵拉4 h和6 h的MyoD和Myf5的表达与对照组相比未有显著变化(P>0.05);3)牵拉结束即刻:15%拉伸度,牵拉4 h和6 h的Akt、P38、ERK和JNK的活性高于对照组(P<0.01),5%拉伸度,牵拉4 h和6 h的Akt、P38、ERK和JNK的活性对照组相比未有显著变化(P>0.05)。牵拉结束24 h后:15%拉伸度,牵拉4 h和6 h的Akt、P38、ERK、JNK活性增加(P<0.05),牵拉结束24 h后:5%拉伸度,牵拉4 h和6 h的Akt、P38、ERK、JNK活性未有显著变化。结论:15%拉伸度的机械牵拉能促进骨骼肌卫星细胞的增殖,这可能是通过激活Akt、P38、ERK、JNK这些通路实现的。Objective:To investigate the effects and mechanisms of different traction modes on the proliferation of rat skeletal muscle satellite cells.Methods:Satellite cells were isolated and purified by two-step enzymatic digestion and differential adherence centrifugation.The third generation satellite cells were collected and a Flex cell pulling system was used to stretch the cells at a frequency of 1 Hz,and 5%and 15%stretches for 4 h and 6 h,respectively.The CCK-8 was used to detect the proliferation of satellite cells at 24 hours after the stretch;the activity of Akt,P38,ERK and JNK was detected immediately after the stretch and at 24 hours after the stretch,and the expression of Myf5 and MyoD was also detected at 24 hours after the stretch.Result:1)Compared with the control group(no stretch,the same below),15%stretch for 4 h or 6 h was increased the proliferation of skeletal muscle satellite cell(P<0.01),but the 5%stretch for 4 h or 6 h had was no significant proliferation(P>0.05);2)A higher protein expression in Myf5 was observed after 15%stretch for 6 h than that of the control group(P<0.01),and the protein expression of MyoD at 4 h and 6 h was both higher than that of the control group(P<0.01).However,no significant difference was observed in the expression of MyoD and Myf5 after 5%stretch for 4 h or 6 h compared with the control group(P>0.05);3)Immediately and 24 hours after traction:The activities of Akt,P38,ERK and JNK after 15%stretch for 4 h or 6 h were higher than those in the control group(P<0.01;P<0.05).However,no significant difference was observed in the activities of Akt,P38,ERK and JNK after 5%stretch for 4 h or 6 h against the control group.There was no significant change(P>0.05).Conclusion:Mechanical traction with 15%stretch can promote the proliferation of skeletal muscle satellite cells,which may be achieved by activating Akt,P38,ERK,JNK pathways.

关 键 词：机械牵拉 肌卫星细胞 增殖 信号通路 

分 类 号：G804.2[文化科学—运动人体科学]