富血小板血浆结合周期性机械牵拉对C2C12肌管萎缩的影响及机制探讨

关键词：富血小板血浆  C2C12骨骼肌细胞  周期性机械牵拉  分化  肌肉萎缩  肌少症

Keywords:platelet-rich plasma  C2C12 skeletal muscle cells  periodic mechanical stretch  differentiation  muscle atrophy  sarcopenia

摘要：

      摘要 目的：研究富血小板血浆（platelet-rich plasma，PRP）结合周期性机械牵拉对C2C12肌管萎缩的影响及机制。 方法：通过蛋白质印记（Western Blot，WB）检测MyoD、MyoG的表达选取最适PRP浓度。随后探究PRP结合周期性机械牵拉（Str）对C2C12细胞的促分化作用；用TNF-α诱导C2C12肌管萎缩，改良吉姆萨染色验证模型；为探究2%PRP结合周期性机械牵拉对C2C12肌管萎缩的作用及机制，将实验分为CON组、TNF-α组、TNF-α+2%PRP组、TNF-α+2%PRP+Str组，并加入抑制剂，检测MuRF-1、Fbx32及Akt、p-Akt、p70、p-p70表达变化。 结果：2% PRP组MyoD、MyoG表达含量明显增加；2% PRP+Str组MyoD、MyoG表达含量增加显著；萎缩实验中，改良吉姆萨染色显示，TNF-α组肌管明显萎缩，TNF-α+2% PRP组可改善肌管萎缩现象；TNF-α+2% PRP+Str组MuRF-1、Fbx32表达含量下降；使用抑制剂干预后，PRP结合周期性机械牵拉的抗萎缩作用被抑制。 结论：2% PRP结合周期性机械牵拉可促进C2C12细胞分化，改善肌管萎缩，通过Akt/p70信号通路发挥作用。

Abstract:

      Abstract Objective： To study effects and mechanism of platelet-rich plasma combined with periodic mechanical stretch on C2C12 myotube atrophy. Method： The expression of MyoD and MyoG was detected by Western Blot（WB） to confirm the optimal concentration of PRP； Subsequently, the pro-differentiation effect of PRP and periodic mechanical stretch （Str） on C2C12 cells was explored； C2C12 cells were induced to shrink by TNF-α，and the models were verified by Modified Giemsa staining. To explore the effect and mechanism of 2%PRP combined with periodic mechanical stretching in improving the atrophy of C2C12 cells， the experiments were divided into control group， TNF-α group， TNF-α+2%PRP group， TNF-α+2%PRP+Str group. The changes of MuRF-1， Fbx32 and Akt， p-Akt， p70 and p-p70 were detected by adding the inhibitors. Result： The expression levels of MyoD and MyoG in the 2% PRP group and the 2% PRP+Str group were significantly increased. In the anti-atrophy experiment， the modified Giemsa staining showed the myotubes in the TNF-α group were significantly atrophied， and the TNF-α+ 2% PRP group can improve myotube atrophy. The expressions of MuRF-1 and Fbx32 were decreased in the TNF-α+2% PRP+Str group. After GDC0068 intervention， the anti-atrophic effect of PRP combined with periodic mechanical stretch was inhibited. Conclusion： 2% PRP combined with periodic mechanical stretch can effectively promote the differentiation of C2C12 cells， improve myotube atrophy， and play a role through the Akt/p70 signaling pathway.