血小板源性生长因子受体α参与成纤维生长因子对辐射诱导人骨髓间充质干细胞损伤的修复作用  

PDGFRα Participates in Basic Fibroblast Growth Factor-mediated Recovery of Human Bone Marrow Mesenchymal Stem Cell Proliferation and Osteogenic Differentiation after Irradiation

作　　者：代凯[1] 杨智[1] 徐双年[1] 张建民[1] 陈洁平[1] 

机构地区：[1]第三军医大学西南医院血液科,重庆400030

出　　处：《中国实验血液学杂志》2015年第6期1709-1715,共7页Journal of Experimental Hematology

基　　金：国家自然基金面上项目(39870046;81270605;30971066;81470324);第三军医大学临床与基础重大基金(2012XLC03);重庆市自然基金(2008BA5001);重庆市研究生教育改革项目(yjg123114);军队重大医学科研基金(BWS13C018)

摘　　要：目的:探讨碱性成纤维生长因子(basic fibroblast growth factor,b FGF)对辐射诱导人骨髓间充质干细胞(human bone marrow mesenchymal stem cell,h BMMSC)增殖、成骨分化损伤后的修复作用及所涉及的可能机制。方法:h BMMSC经0、6、12 Gy X射线辐照后,流式细胞术、cell counting kit-8(CCK-8)、Western blot法和茜素红染色法检测辐射对h BMMSC凋亡、增殖、成骨分化的影响;0、1、5、10、20 ng/ml浓度b FGF分别作用于辐射h BM M SC,探索抗辐射保护作用最佳浓度;Western blot法检测血小板源性生长因子受体α(platelet derived grow th factor receptor alpha,PDGFRα)蛋白表达动态变化。结果:辐射后h BM M SC增殖、成骨分化能力下降,b FGF能够修复h BMMSC增殖、成骨分化损伤(P<0.05),5 ng/ml浓度作用最显著;仅辐射后早期(24 h)12 Gy组h BMMSC凋亡高于0 Gy组(P<0.05),其余时间点辐射组与未辐照组细胞凋亡无差异(P>0.05)。辐射后h BMMSC血小板源性生长因子受体-α表达明显下调,b FGF作用后PDGFRα表达上调。结论:辐射对h BMMSC凋亡无显著影响,b FGF对h BMMSC辐射损伤有修复作用,促进其增殖和成骨分化功能的恢复;h BMMSC增殖、成骨分化损伤与辐射诱导PDGFRα表达下调相关。PDGFRα参与b FGF对h BMMSC辐射损伤的修复。Objective:To explore the effects of basic fibroblast growth factor(bFGF) on human bone marrow mesenchymal stem cell(hBMMSC) damaged by irradiation and its underlying mechanisms.Methods:hBMMSC was irradiated with 0,6,12 Gy X ray,then flow cytometry,cell counting kit-8(CCK-8),Western blot and alizarin red staining were used to detect the effects of X ray on apoptosis proliferation and osteogenic differentiation of hBMMSC;0,1,5,10,20 ng/ml bFGF was added to hBMMSC irradiated with X ray for selecting the suitable bFGF reaction concentration;then the Western blot was used to detect the expression of PDGFRa so as to evaluate whether the expression of PDGFRa participated in bFGF-mediated recovery of hBMMSC proliferation and osteogenic differentiation after irradiation.Results:The proliferation and osteogenic differentiation of hBMMSC decreased remarkably after irradiation.bFGF promoted the recovery of proliferation and osteogenic differentiation of irradiated hBMMSC compared with untreated irradiated hBMMSC(P <0.05);5 ng/ml bFGF was identified as the optimal concentration.A significant difference in the number of apoptotic cells could be detected only between the 0 Gy group and 12 Gy group at the 24 h time point,while no differences were detected at later time points.Irradiated hBMMSC showed remarkable decrease of PDGFRa expression,while the PDGFRa expression increased after bFGF was added.Conclusion:Irradiation dose not show significant effect on apoptosis of hBMMSC,but the bFGF displays a effect on repairing the irradiation damage of hBMMSC and promotes the recovery of hBMMSC proliferation and osteogenic differentiation.The damage of hBMMSC proliferation and osteogenic differentiation associates with downregulation of PDGFRa expression induced by irrediation.PDGFRa involves in repairing effect of bFGF on irradiation damage of hBMMSC.

关 键 词：辐射损伤 人骨髓间充质干细胞 碱性成纤维生长因子 凋亡 增殖 成骨分化 血小板源性生长因子受体-α 

分 类 号：R818[医药卫生—放射医学]