低强度脉冲式超声波上调细胞周期蛋白D1表达促进骨髓间充质干细胞增殖  

Low-intensity pulsed ultrasound promotes the proliferation of bone marrow mesenchymal stem cells by inducing cyclin D1 up-regulation

作　　者：李志 华永永 张剑权[4] 符真[4] Li Zhi;Hua Yongyong;Zhang Jianquan;Fu Zhen(Ward Three,Department of General Surgery,Taihe Hospital,Hubei University of Medicine,Shiyan 442000,Hubei Province,China;Hubei Key Laboratory of Embryonic Stem Cell Research,Taihe Hospital,Hubei University of Medicine,Shiyan 442000,Hubei Province,China;Taihe Hospital Affiliated to Xi’an Jiaotong University Health Science Center,Shiyan 442000,Hubei Province,China;Department of Hepatobiliary Surgery,Haikou Hospital Affiliated to Xiangya School of Medicine,Central South University,Haikou 570000,Hainan Province,China)

机构地区：[1]湖北医药学院附属太和医院普外科三病区,湖北省十堰市442000 [2]湖北医药学院附属太和医院胚胎干细胞研究湖北省重点实验室,湖北省十堰市442000 [3]西安交通大学医学部附属太和医院,湖北省十堰市442000 [4]中南大学湘雅医学院附属海口医院肝胆外科,海南省海口市570000

出　　处：《中国组织工程研究》2022年第13期2006-2011,共6页Chinese Journal of Tissue Engineering Research

基　　金：海南省重点科技计划项目(ZDXM2015082),项目负责人:张剑权;海南省自然科学基金项目(817382),项目负责人:符真。

摘　　要：背景:前期研究发现低强度脉冲式超声波可以促进人骨髓间充质干细胞增殖。目的:探讨FAK/PI3K/Akt信号通路在低强度脉冲式超声波促进人骨髓间充质干细胞增殖中的作用机制。方法:将购买的商品化人骨髓间充质干细胞复苏培养至第4代,分为0 mW/cm^(2)超声波组(对照组)、50 mW/cm^(2)超声波组、50 mW/cm^(2)超声波+PF-562271组,其中50 mW/cm^(2)超声波+PF-562271组在超声处理前加入1μL FAK/PI3K/Akt信号通路抑制剂PF-562271进行预处理。每隔24 h超声刺激1次,每次5 min,刺激3次后采用CCK-8试剂盒检测细胞增殖情况,流式细胞仪检测细胞周期变化,Western blot法检测FAK及其下游靶基因和细胞周期蛋白D1的表达。结果与结论:①与对照组相比,50 mW/cm^(2)超声波组细胞增殖能力明显增强(P﹤0.05),使用PF-562271抑制FAK/PI3K/Akt信号通路后,细胞增殖能力明显降低(P﹤0.001);②与对照组相比,50 mW/cm^(2)超声波组FAK、p-FAK、PI3K、p-PI3K、Akt、p-Akt和细胞周期蛋白D1的表达均上调(P﹤0.05);与50 mW/cm^(2)超声波组相比,50 mW/cm^(2)超声波+PF-562271组上述指标明显降低(P﹤0.05);③与对照组相比,50 mW/cm^(2)超声波组G_(1)期细胞比例减少(P=0.026),S期细胞比例增多(P=0.002);与50 mW/cm^(2)超声波组相比,50 mW/cm^(2)超声波+PF-562271组G_(1)期细胞比例升高(P=0.023),S期细胞比例降低(P=0.035);④结果表明,低强度脉冲式超声波可能通过FAK/PI3K/Akt信号通路上调细胞周期蛋白D1促进人骨髓间充质干细胞的增殖。BACKGROUND:Previous studies found that low-intensity pulsed ultrasound can promote the proliferation of human bone marrow mesenchymal stem cells.OBJECTIVE:To investigate the mechanism of low-intensity pulsed ultrasound on the proliferation of human bone marrow mesenchymal stem cells via the FAK/PI3K/Akt signaling pathway.METHODS:The purchased commercial human bone marrow mesenchymal stem cells were resuscitated and cultured to the fourth generation,and they were divided into 0 mW/cm^(2) ultrasound group(control group),50 mW/cm^(2) ultrasound group,and 50 mW/cm^(2) ultrasound+PF-562271 group.The 50 mW/cm^(2) ultrasound+PF-562271 group was pretreated with 1μL FAK/PI3K/Akt signaling pathway inhibitor PF-562271 before ultrasound treatment,once every 24 hours,5 minutes each time.After three stimuli,cell proliferation was detected by Cell Counting Kit-8 and the cell cycle was detected by flow cytometry.Western blot assay was used to detect the expression of FAK and its downstream target gene and cyclin D1.RESULTS AND CONCLUSION:(1)Compared with the control group,the cell proliferation ability was significantly enhanced in the 50 mW/cm^(2) ultrasound group(P<0.05).After inhibiting the FAK/PI3K/Akt signaling pathway using PF-562271,the cell proliferation ability was significantly reduced(P<0.001).(2)Compared with the control group,the expression levels of FAK,p-FAK,PI3K,p-PI3K,Akt,p-Akt and cyclin D1 were all up-regulated in the 50 mW/cm^(2) ultrasound group(P<0.05).Compared with the 50 mW/cm^(2) ultrasound group,the above indicators were significantly reduced in the 50 mW/cm^(2) ultrasound+PF-562271 group(P<0.05).(3)Compared with the control group,the proportion of cells in the G_(1) phase decreased in the 50 mW/cm^(2) ultrasound group(P=0.026),and the proportion of cells in the S phase increased(P=0.002).Compared with the 50 mW/cm^(2) ultrasound group,the proportion of cells in G_(1) phase increased(P=0.023),and the proportion of cells in S phase decreased in the 50 mW/cm^(2) ultrasound+PF-562271 group(P=0.035).(4)The

关 键 词：干细胞 人骨髓间充质干细胞 低强度脉冲式超声波 增殖 通路 细胞周期 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]