冲击波干预后骨髓间充质干细胞转化生长因子-β_1和磷酸化细胞外信号调节激酶表达的动态观察

Dynamic observation of expression of TGF-β_1 and phosphorylate exracellular signal-regulated kinases in human mesenchymal stem cells after extracoporeal shock wave intervention.

目的动态观察体外冲击波干预对骨髓间充质干细胞(hMSCs)表达骨细胞生长因子和酶联信号分子的影响,探讨其诱导成骨的机制。方法将5kV、100频次的体外冲击波作用于第1代骨髓间充质干细胞,采用免疫细胞化学法隔代观察(P3-P11)转化生长因子-β1(TGF-β1)和磷酸化细胞外信号调节激酶(pERK)的表达,计数阳性率。结果5kV的体外冲击波对骨髓间充质干细胞增殖的影响显著;干预后的P3代hMSCs胞浆内即出现黄染颗粒,pERK1/2弱阳性;对照组仅P9代个别hMSCs阳性(P<0.01)。干预组P9代hMSCs中pERK1/2活跃,TGF-β1出现强阳性表达(72.2%),随后开始下降;与对照组比较差异有显著性(P<0.01)。结论体外冲击波干预后pERK1/2被激活,TGF-β1参与了hMSCs增殖后期ERKs酶联信号活动。

Objective To discuss the osteogenetic mechanism of Extracorporeal shock wave（ESW, dynamic expression of TGF-β1 and phosphorylate extracellular signal-regulated kinases （ pERKs ） mesenchymal stem cells after ESW intervention. Methods After success in marrow aspiration and isolation, an optimal dose of 5kV and 100 times of ESW was applied to hMSCs. The expression of pERK and TGF-β1 were examined by immunocytochemical staining. Results pERK shouved weak positive expressina, which mediated he, cascades of mitogen - activated protein kinases signaling pathway was activated in hMSCs of p3 in exposure group, and earlier than the expression of TGF-β1. At the same interval,the expression of pERK and TGF-β1 in ESW group were significantly lower than that of control group （ P 〈 0.01 ）. pERK1/2 of p9） in Esw group was activated and TGF-β1 shoued strong positive expression（72.2% ） ,which began to decrease,with signigicant difference（ P 〈 0.01 ） Conclusion ESW of 5kV and 100 times indirectly activates signaling cascades of pERKs, which could participate the process of ESW accelerating osteogenesis and be associated with the appearance of TGF-β1.