FGFR2功能增强对软骨内成骨作用的机制研究  

The study of the mechanism in the endochondral ossification of bone mesenchymal stem cells in mice by continued enhanced function of fibroblast growth factor typeⅡ receptor mutation

作　　者：陈鹏[1] 张凡喜[1] 周玉峰[1] 张波[2] 

机构地区：[1]解放军第324医院神经外科,重庆400020 [2]第三军医大学大坪医院野战外科研究所,重庆400042

出　　处：《重庆医学》2015年第8期1009-1011,1016,共4页Chongqing medicine

基　　金：国家973计划重点基础研究发展计划基金资助项目(2011CB964701)

摘　　要：目的利用模拟人Apert综合征的FGFR2S252W/+小鼠模型,研究ERK信号通路在成纤维细胞生长因子受体2(FGFR2)功能持续增强对软骨内成骨过程的影响。方法动物繁殖、鉴定后分为FGFR2功能获得突变型和同窝野生型。于出生后6周获取骨髓间充质干细胞(BMSCs)进行体外培养及2代BMSCs成软骨诱导,对ERK信号蛋白检测,并比较Col2、Col10、OC、OP基因的表达。加入ERK信号通路阻滞剂PD98059后再次培养观察相应基因的表达。胚胎骨体外培养技术比较ERK信号通路在FGFR2功能持续增强对软骨内成骨过程的影响。结果在体外BMSCs成软骨诱导后观察发现,FGFR2功能突变小鼠BMSCs表达总ERK蛋白量无明显变化,但磷酸化增强。FGFR2S252W/+小鼠BMSCs表达Col2、Col10较野生型弱,但OC、OP基因强于野生型小鼠。加入ERK信号通路阻滞剂PD98059培养后,对Col2、Col10基因影响不大,但OC、OP表达量增高。并且在胚胎骨体外培养过程中,发现PD98059治疗组能纠正FGFR2功能持续增强所致软骨内成骨发育障碍。结论 ERK信号通路是FGFR2下游影响软骨内成骨的关键通路,特别对软骨内成骨后期成骨过程影响巨大。其信号通路阻滞剂对软骨内成骨发育障碍有一定救治作用。Objective To study the role of ERK signal pathway in the endochondral ossification of bone mesenchymal stem cells ,and to explore the mechanism of ERK signal pathway in persistent enhanced FGFR2 function on development of mice BMSCs by a knock‐in mouse model with the FGFR2S252W/＋ .Methods Mice with neo‐FGFR2 gain‐of‐function mutation were mated with EII‐Cre mice .The genotype of generation mice were identified by PCR and divided into wild type group and mutant type group ac‐cording to their genotype .6 week‐old mice were sacrificed to receive bone mesenchymal stem cells .The western blot was used to compare the level of P‐ERK and ERK and the RT‐PCR was applied to detect the genes of Col2 ,Col10 ,OC ,OP in chondrogenic dif‐ferentiation medium of BMSCs .Then ,treatment of cultured BMSCs with PD98059 ,compare the changes of genes and utilize the in vitro culture of long bones detect the role of ERK signal pathway in the endochondral ossification by FGFR2 mutant .Results We successfully derive BMSCs from FGFR2S252W/＋ mutant mice and found the activity of ERK signal pathway of FGFR2S252W/＋ was en‐hanced .After been cultured in chondrogenic differentiation medium ,the expressions of the BMSCs mRNA of Col2 ,Col10 from mu‐tant group were decreased ,while the expressions of OC ,OP were increased .Those OC ,OP genes levels showed an increased treated by PD98059 .Using in vitro culture of long bones ,we found the retardation of total length growth of long bones has been rescued by PD98059 treatment ,suggesting that ERK signal pathways was responsible for the retarded long bone development in FGFRS252W/＋mice .Conclusion The results indicate these effects are mediated by the ERK signal pathway .Furthermore ,the retardation of long bones has been recued by PD98059 treatment ,suggesting that ERK signal pathway is responsible for the retarded long bone devel‐opment in FGFR2S252W/＋ mice .

关 键 词：成纤维细胞生长因子受体2 ERK信号通路 骨髓间充质干细胞 软骨内成骨 软骨分化 

分 类 号：R336[医药卫生—人体生理学]