中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
7期
1402-1405
,共4页
陈鹏%张凡喜%周玉峰%张波
陳鵬%張凡喜%週玉峰%張波
진붕%장범희%주옥봉%장파
成纤维细胞生长因子受体2%p38信号通路%骨髓间充质干细胞%软骨内分化
成纖維細胞生長因子受體2%p38信號通路%骨髓間充質榦細胞%軟骨內分化
성섬유세포생장인자수체2%p38신호통로%골수간충질간세포%연골내분화
Fibroblast growth factor receptor 2%p38 signal pathway%Bone mesenchymal stem cells%Chondrogenic differentiation
目的 基因敲入技术建立成纤维细胞生长因子受体2(FGFR2)功能持续增强小鼠模型(FGFR2S252W/+),观察p38信号通路在FGFR2功能持续增强对软骨内成骨过程的影响.方法 获取出生后6周小鼠骨髓间充质干细胞(BMSCs)进行体外培养并进行成软骨诱导.Western blot检测pβ8信号蛋白,逆转录-聚合酶链反应(RT-PCR)比较野生型及突变型Ⅱ型胶原(Col2)、X型胶原(Col10)、OC、OP基因表达,加入p38信号通路阻滞剂SB203580后再次比较相关基因表达.体外胚胎骨培养观察pβ8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 体外BMSCs成软骨诱导后,FGFR2功能突变小鼠BMSCs表达p38蛋白磷酸化增强,表达Col2、Col10减弱,但OC、OP基因表达强于野生型;加入p38信号通路阻滞剂SB203580后,BMSCs基因表达量明显增高,表达Col2、Col10为原来1.30±0.07、1.94±0.13,表达OC、OP为原来1.97±0.17、1.50±0.10.胚胎骨体外培养可见SB203580治疗能纠正软骨内成骨发育障碍,使胫骨的总长度及钙化组织长度明显增长.结论 FGFR2下游p38信号通路对软骨内成骨过程影响巨大,其信号通路阻滞剂对软骨内成骨发育障碍有救治作用.
目的 基因敲入技術建立成纖維細胞生長因子受體2(FGFR2)功能持續增彊小鼠模型(FGFR2S252W/+),觀察p38信號通路在FGFR2功能持續增彊對軟骨內成骨過程的影響.方法 穫取齣生後6週小鼠骨髓間充質榦細胞(BMSCs)進行體外培養併進行成軟骨誘導.Western blot檢測pβ8信號蛋白,逆轉錄-聚閤酶鏈反應(RT-PCR)比較野生型及突變型Ⅱ型膠原(Col2)、X型膠原(Col10)、OC、OP基因錶達,加入p38信號通路阻滯劑SB203580後再次比較相關基因錶達.體外胚胎骨培養觀察pβ8信號通路在FGFR2功能持續增彊對軟骨內成骨過程的影響.結果 體外BMSCs成軟骨誘導後,FGFR2功能突變小鼠BMSCs錶達p38蛋白燐痠化增彊,錶達Col2、Col10減弱,但OC、OP基因錶達彊于野生型;加入p38信號通路阻滯劑SB203580後,BMSCs基因錶達量明顯增高,錶達Col2、Col10為原來1.30±0.07、1.94±0.13,錶達OC、OP為原來1.97±0.17、1.50±0.10.胚胎骨體外培養可見SB203580治療能糾正軟骨內成骨髮育障礙,使脛骨的總長度及鈣化組織長度明顯增長.結論 FGFR2下遊p38信號通路對軟骨內成骨過程影響巨大,其信號通路阻滯劑對軟骨內成骨髮育障礙有救治作用.
목적 기인고입기술건립성섬유세포생장인자수체2(FGFR2)공능지속증강소서모형(FGFR2S252W/+),관찰p38신호통로재FGFR2공능지속증강대연골내성골과정적영향.방법 획취출생후6주소서골수간충질간세포(BMSCs)진행체외배양병진행성연골유도.Western blot검측pβ8신호단백,역전록-취합매련반응(RT-PCR)비교야생형급돌변형Ⅱ형효원(Col2)、X형효원(Col10)、OC、OP기인표체,가입p38신호통로조체제SB203580후재차비교상관기인표체.체외배태골배양관찰pβ8신호통로재FGFR2공능지속증강대연골내성골과정적영향.결과 체외BMSCs성연골유도후,FGFR2공능돌변소서BMSCs표체p38단백린산화증강,표체Col2、Col10감약,단OC、OP기인표체강우야생형;가입p38신호통로조체제SB203580후,BMSCs기인표체량명현증고,표체Col2、Col10위원래1.30±0.07、1.94±0.13,표체OC、OP위원래1.97±0.17、1.50±0.10.배태골체외배양가견SB203580치료능규정연골내성골발육장애,사경골적총장도급개화조직장도명현증장.결론 FGFR2하유p38신호통로대연골내성골과정영향거대,기신호통로조체제대연골내성골발육장애유구치작용.
Objective To study the role of p38 signal pathway in the endochondral ossification of bone mesenchymal stem cells (BMSCs) of wild type mice and mutant type [fibroblast growth factor receptor 2 (FGFR2)S252w/+),and to explore the mechanism of p38 signal pathway in persistent enhanced FGFR2 function on development of mice BMSCs by a knock-in mouse model with the FGFR2S252w/+.Methods The BMSCs were isolated from 60week-old mice.Western blotting was used to compare the level of P-p38 and p38,and the reverse transcription-polymerase chain reaction (RT-PCR) was applied to detect the genes of Col2,Col10,OC and OP in chondrogenic differentiation medium of BMSCs.The cultured BMSCs were treated with SB203580,the expression of genes was compared,and the in vitro culture of long bones was utilized to detect the role of p38 signal pathway in the endochondral ossification by FGFR2 mutant.Results The activity of p38 signal pathway of FGFR2S252w/+ was enhanced.After culture in chondrogenic differentiation medium,the mRNA expression of collagentype Ⅱ (Col2) and collagentype X (Col10) in BMSCs from mutant group was decreased,and that of OC and OP was increased.After treatment with SB203580,the expression of these genes was increased,SB203580 group express Col2,Col10 were 1.30 ±0.07 and 1.94 ± 0.13 times the MT group,expres OC,OP were 1.97 ± 0.17,1.50 ± 0.10 times the MT group.Using in vitro culture of long bones,it was found the retardation of total length growth of long bones was rescued by SB203580 treatment,suggesting that p38 signal pathways was responsible for the retarded long bone development in FGFR2S252w/+ mice.Conclusion The results indicated that these effects are mediated by the p38 signal pathway.Furthermore,the retardation of long bones has also been rescued by SB203580 treatment,suggesting that p38 signal pathway was responsible for the retarded long bone development in FGFR2S252W/+ mice.
