冲击波通过P2X7受体激活人骨髓间充质干细胞的p38 MAPK通路  

Shockwave-induced cellular ATP release activates P2X7 receptors which elicited osteogenic differentiation of human mesenchymal stem cells by the phosphorylation of p38MAPK

作　　者：侯向锋[1] 王成学[1] 赵毅[1] 郑学清[2] 黄玉龙[1] 张海鹏[1] 钟子龙 于铁成[1] 

机构地区：[1]吉林大学白求恩第一医院创伤骨科,吉林长春130021 [2]吉林大学白求恩第一医院碎石中心 [3]吉林大学白求恩第一医院手术室

出　　处：《中国老年学杂志》2014年第6期1540-1543,共4页Chinese Journal of Gerontology

基　　金：国家自然科学基金项目(No.81172183)

摘　　要：目的探讨体外冲击波是否通过促使ATP向细胞外释放激活P2X7受体信号通路并诱导人骨髓间充质干细胞(hMSCs)p38 MAPK激酶活化。方法用即时RT-PCR检测P2X7受体mRNA的表达;通过免疫印迹法评估P2X7受体和p38 MAPK激酶的表达;用ATP水解酶、P2X7受体的siRNA、p38 MAPK激酶抑制剂以及P2受体的抑制剂评估ATP释放、P2X7受体在冲击波诱导hMSCs p38 MAPK激酶磷酸化中的作用。结果 (能量密度0.18 mJ/mm2,作用0、50、100或150次)冲击波可引起细胞内的ATP(~6.9μmol/L)向外释放,冲击波(能量密度0.18 mJ/mm2,作用0、50、100或150次)和细胞外的ATP(0.1~1μmol/L)能够促使p38 MAPK激酶活化。用ATP水解酶消除细胞外ATP、用siRNA或抑制剂抑制P2X7受体抑制p38 MAPK激酶的活化。结论冲击波引起的细胞内ATP向外释放,能够有效激活P2X7受体传导信号通路,引起hMSCs向成骨细胞的p38MAPK激酶活化,这可能是冲击波疗法的机制所在。Objective To investigate whether this effect could be due to ATP release-induced the activation of p38MAPK of human mesenchymal stem cells( hMSCs). Methods Cultured bone marrow-derived hMSCs were subjected to shockwave treatment and ATP release was assessed. Expressions of P2X7 receptors mRNA were determined by real-time RT-PCR. Western blot was used to assess the expression of P2X7 receptors and the activation of p38MAPK. P2X7-siRNA,apyrase,P2 receptor antagonists were used to evaluate the roles of ATP release,and P2X7 receptors in shockwave-induced osteogenic hMSCs p38 MAPK signaling. Results Shockwave treatment released significant amounts( ~ 6. 9 μmol / L) of ATP from hMSCs. Shockwaves and exogenous ATP induced p38 MAPK activation. Removal of ATP with apyrase,targeting of P2X7 receptors with P2X7-siRNA or selective antagonist prevented the activation of p38MAPKof hMSCs. Conclusions Shockwaves release cellular ATP that activates P2X7 receptors and downstream signaling events that cause osteogenic the activation of p38MAPKof hMSCs. Shockwave therapy might promote bone healing through P2X7 receptor signaling,which contributes to the activation of p38MAPK of hMSCs.

关 键 词：冲击波 人骨髓间充质干细胞 ATP P2X7受体 p38 MAPK 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]