| 刘秀娟,张念云,王 斌,孙 飙,徐 妍.脂肪间充质干细胞注射对离心运动大鼠肌肉修复中肌肉抑制素信号通路的影响机制[J].中国康复医学杂志,2021,(1):16~22 |
| 脂肪间充质干细胞注射对离心运动大鼠肌肉修复中肌肉抑制素信号通路的影响机制 点此下载全文 |
| 刘秀娟 张念云 王 斌 孙 飙 徐 妍 |
| 南京体育学院运动健康学院运动人体科学系，江苏省南京市玄武区灵谷寺路8号，210014 |
| 基金项目：国家自然科学基金青年项目（31900844） |
| DOI：10.3969/j.issn.1001-1242.2021.01.004 |
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| 摘要： |
| 摘要
目的：通过肌肉注射异体脂肪间充质干细胞，观察大鼠骨骼肌肌肉抑制素（myostatin，MSTN）信号通路及肌纤维超微结构的变化，探讨脂肪间充质干细胞在离心运动后骨骼肌修复过程中的作用机制。
方法：8周龄雄性SD大鼠，进行一次性离心运动后，在大鼠的左腿腓肠肌外侧头注射生理盐水（NS），右腿腓肠肌外侧头注射脂肪间充质干细胞（ASCs），然后，随机分为运动后1天组（D1）、运动后3天组（D3）、运动后7天组（W1）和运动后14天组（W2）。分别在相应的时间点将大鼠处死，收集样本。采用ELISA测定血清肌酸激酶（CK）、骨骼肌肌钙蛋白I（sTnI）、MSTN、卵泡抑素（follistatin，FST）的含量；实时荧光定量PCR测定骨骼肌MSTN、激活素A受体ⅡB（ACVR2B）、FST mRNA相对表达量。Western Blot测定骨骼肌MSTN、ACVR2B、FST、信号转导蛋白smad2/3磷酸化（p-smad2/3）蛋白的表达。
结果：与D1组比较，W1、W2组血清CK水平显著降低（P<0.05），D3和W1组血清sTnI显著升高（P<0.05）；与D1组比较，W1组血清MSTN水平显著降低（P<0.05），但W2组血清MSTN水平显著升高（P<0.05）；与D1组比较，W2组血清FST水平显著升高（P<0.05）。与NS组相比，ASCs组MSTN mRNA 相对表达量在时间点D3显著降低（P<0.05），在时间点W2极显著降低（P<0.01）；ACVR2B和FST mRNA相对表达量与NS组相比无显著变化；与NS组相比，ASCs组MSTN蛋白表达在时间点D1和D3均显著降低（P<0.05）；ACVR2B蛋白表达在时间点D1、D3、W1均显著升高（P<0.05），在时间点W2却显著降低（P<0.05）；与NS组相比，smad2/3的磷酸化水平在时间点W2显著降低（P<0.05）。
结论：离心运动后，肌注异体脂肪间充质干细胞对MSTN的影响可能比对FST的影响大；肌注异体脂肪间充质干细胞可能通过影响MSTN下游信号通路来促进离心运动后骨骼肌的再生修复。 |
| 关键词：离心运动 脂肪间充质干细胞 骨骼肌 肌肉抑制素 |
| The role of myostatin signaling pathway in the improvement of muscle repair by injection of adipose mesenchymal stem cells in rats after eccentric exercise Download Fulltext |
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| Department of Sports Science，School of Kinesiology，Nanjing Sports Institute，Nanjing，210014 |
| Fund Project: |
| Abstract: |
| Abstract
Objective： To observe the changes of myostatin signaling pathway and muscle fiber ultrastructure by injection of adipose mesenchymal stem cells (ASCs) in rats after a single eccentric exercise， to investigate the mechanism of ASCs injection in skeletal muscle injury repair after eccentric exercise.
Method： After a one-off eccentric exercise in 8-week-old male SD rats， normal saline was injected into the gastrocnemius muscle in the left leg， and ASCs were injected into the gastrocnemius muscle of the right leg. The rats were randomly divided into four groups： one day(D1)，three days(D3)， seven days(W1) and fourteen days(W2) post exercise. The rats were killed at the corresponding time points. The content of CK、sTnI、myostatin（MSTN）and follistatin（FST）in serum was determined by ELISA method. Real-time fluorescence quantitative PCR was used to detect the expressions of MSTN、ACVR2B and FST mRNA in skeletal muscle. The expressions of MSTN、ACVR2B、FST and p-smad2/3 protein in skeletal muscle were detected by Western Blot.
Result： Compared with group D1， the serum CK contents in group W1 and group W2 were significantly decreased (P<0.05)， the serum sTnI content in group D3 and group W1 was remarkably increased (P<0.05)， the serum myostatin level in group W1 was significantly decreased (P<0.05)， but which was remarkably increased (P<0.05) in group W2. Compared with group D1， the serum FST level in group W2 was significantly increased (P<0.05). Compared with group NS， relative expression quantity of MSTN mRNA in group ASCs was significantly decreased at time point D3 (P<0.05)，which was extremely significantly down-regulated at time point W2 (P<0.01). However， relative expression quantities of ACVR2B and FST mRNA had no significant difference between the two groups. Compared with group NS， the expression of MSTN protein was significantly decreased at time point D1 and D3 (P<0.05)， the expression of ACVR2B protein was remarkably increased at time point D1、D3 and W1(P<0.05)， while significantly decreased at time point W2 (P<0.05). Compared with group NS， the phosphorylation of smad2/3 was significantly decreased at the time point W2 in group ASCs (P<0.05).
Conclusion： After eccentric exercise， the effect of allogeneic adipose mesenchymal stem cells on MSTN may be greater than on FST. Allogeneic adipose mesenchymal stem cells injected intramuscularly may improve the regeneration and repair of skeletal muscle after eccentric exercise through affecting the downstream signaling pathway of MSTN. |
| Keywords:eccentric exercise adipose mesenchymal stem cells skeletal muscle myostatin |
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