中华创伤骨科杂志
中華創傷骨科雜誌
중화창상골과잡지
CHINESE JOURNAL OF ORTHOPAEDIC TRAUMA
2009年
4期
362-365
,共4页
唐欣%徐忠世%杨述华%陈雨辰%李奇%余从年%杨操%李进%许伟华
唐訢%徐忠世%楊述華%陳雨辰%李奇%餘從年%楊操%李進%許偉華
당흔%서충세%양술화%진우신%리기%여종년%양조%리진%허위화
成纤维细胞生长因子%软骨%寡核苷酸序列分析%骨髓细胞%转染
成纖維細胞生長因子%軟骨%寡覈苷痠序列分析%骨髓細胞%轉染
성섬유세포생장인자%연골%과핵감산서렬분석%골수세포%전염
Fasic fibroblast growth factor%Cartilage%Oligonucleotide array sequence analysis%Bone marrow cells%Transfection
目的 检测并分析促分裂原活化蛋白激酶(MAPK)信号通路中成纤维细胞生长因子(FGF)在软骨发育过程中的基因表达规律;通过细胞培养观察FGF基因对骨髓基质干细胞(BMSCs)生长特性的影响. 方法用基因芯片技术建立妊娠胎鼠肢芽软骨发育过程的基因表达谱,分析MAPK信号通路中碱性成纤维细胞生长因子(brGF)在软骨发育过程中的基因表达规律.构建bFGF质粒并转染至培养的BMSCs中,用MTT法、免疫组织化学、HE染色、RT-PCR及酶联免疫吸附法检测bFGF基因转染BMSCs的效果及产物表达. 结果 MAPK信号通路中的FGF在软骨发育过程中的软骨形成关键期表达显著上调,并启动MAPK信号通路,促进软骨形成.bFGF基因转染的BMSCs生长活力较强,可以保持2周以上;HE染色显示细胞增殖旺盛,胞核深染;RT-PCR表明有bFGF的基因表达;酶联免疫吸附法检测bFGF表达量高. 结论 FGF能够启动MAPK信号通路从而促进软骨彤成.bFGF质粒转染BMSCs后可促进BMSCs的增殖,细胞有向软骨细胞分化趋势.
目的 檢測併分析促分裂原活化蛋白激酶(MAPK)信號通路中成纖維細胞生長因子(FGF)在軟骨髮育過程中的基因錶達規律;通過細胞培養觀察FGF基因對骨髓基質榦細胞(BMSCs)生長特性的影響. 方法用基因芯片技術建立妊娠胎鼠肢芽軟骨髮育過程的基因錶達譜,分析MAPK信號通路中堿性成纖維細胞生長因子(brGF)在軟骨髮育過程中的基因錶達規律.構建bFGF質粒併轉染至培養的BMSCs中,用MTT法、免疫組織化學、HE染色、RT-PCR及酶聯免疫吸附法檢測bFGF基因轉染BMSCs的效果及產物錶達. 結果 MAPK信號通路中的FGF在軟骨髮育過程中的軟骨形成關鍵期錶達顯著上調,併啟動MAPK信號通路,促進軟骨形成.bFGF基因轉染的BMSCs生長活力較彊,可以保持2週以上;HE染色顯示細胞增殖旺盛,胞覈深染;RT-PCR錶明有bFGF的基因錶達;酶聯免疫吸附法檢測bFGF錶達量高. 結論 FGF能夠啟動MAPK信號通路從而促進軟骨彤成.bFGF質粒轉染BMSCs後可促進BMSCs的增殖,細胞有嚮軟骨細胞分化趨勢.
목적 검측병분석촉분렬원활화단백격매(MAPK)신호통로중성섬유세포생장인자(FGF)재연골발육과정중적기인표체규률;통과세포배양관찰FGF기인대골수기질간세포(BMSCs)생장특성적영향. 방법용기인심편기술건립임신태서지아연골발육과정적기인표체보,분석MAPK신호통로중감성성섬유세포생장인자(brGF)재연골발육과정중적기인표체규률.구건bFGF질립병전염지배양적BMSCs중,용MTT법、면역조직화학、HE염색、RT-PCR급매련면역흡부법검측bFGF기인전염BMSCs적효과급산물표체. 결과 MAPK신호통로중적FGF재연골발육과정중적연골형성관건기표체현저상조,병계동MAPK신호통로,촉진연골형성.bFGF기인전염적BMSCs생장활력교강,가이보지2주이상;HE염색현시세포증식왕성,포핵심염;RT-PCR표명유bFGF적기인표체;매련면역흡부법검측bFGF표체량고. 결론 FGF능구계동MAPK신호통로종이촉진연골동성.bFGF질립전염BMSCs후가촉진BMSCs적증식,세포유향연골세포분화추세.
Objective To explore the gene expression profile of fibmblast growth factor (FGF) in MAPK signaling pathway during entochondrostosis in mice and investigate the effects of transfectiou of pcDNA-bFGF on mice bome marrow stromal cells (BMSCs) in vitro. Methods cDNA microarray technique with 34,000 genes was used to analyze the gene expression profile during entochondrostosis in the limbs of mice embryo from E10 to E14. Pathway analysis of FGF in MAPK signaling pathway was performed with GCOS1.2 software. The recombined expression vector PcDNA-bFGF was constructed and transfected into mice BMSCs by Lipofectaming The phenotype changes of cells were observed by cell energometry, HE stain and immunohistochemical assay under light mi-croscopy, RT-PCR and Elisa. Results The gene expression of FGF in MAPK signaling pathway during the critical phase of chondrogenesis in the limbs of mice embryo at E12 was increased obviously. FGF started the MAPK signaling pathway and promoted chondrogenesis. The phenotypes of BMSCs were verified significantly except the bFGF transfected group. As compared with the vector and blank groups, the bFGF transfected group had a tendency of chondrocyte cytodifferentiation. Conclusions FGF may start the MAPK signaling pathway and promote chondrogenesis. The transfection of bFGF gene into mice BMSCs can help BMSCs differentiate into chondrocytes, which should be further investigated for cartilage tissue engineering.
