中华风湿病学杂志
中華風濕病學雜誌
중화풍습병학잡지
CHINESE JOURNAL OF RHEUMATOLOGY
2008年
5期
325-329
,共5页
唐欣%徐忠世%杨述华%陈雨辰%李奇%余从年%杨操%李进%许伟华%杜靖远
唐訢%徐忠世%楊述華%陳雨辰%李奇%餘從年%楊操%李進%許偉華%杜靖遠
당흔%서충세%양술화%진우신%리기%여종년%양조%리진%허위화%두정원
芯片分析技术%SOX9基因%软骨发生%间质干细胞
芯片分析技術%SOX9基因%軟骨髮生%間質榦細胞
심편분석기술%SOX9기인%연골발생%간질간세포
Microchip analytical procedures%Sry-type high-mobility-group box gene-9%Chondrogenesis%Mesenchymal stem cells
目的 检测并分析SOX9基因在软骨发育过程中的表达规律;构建其质粒并转染至培养的骨髓源基质细胞(BMSCs)中,通过细胞培养观察SOX9基因对BMSCs生长特性的影响,为可能的骨关节炎(OA)基因治疗提供理论基础.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SOX9基因在软骨发育过程中的表达规律;用噻唑蓝(MTT)法、免疫组织化学、苏木素-伊红(HE)染色、反转录-聚合酶链反应(RT-PCR)及酶联免疫吸附试验(ELISA)法检测SOX9基因转染MSCs的效果及产物的表达.结果 SOX9基因在软骨发育过程中的软骨形成关键期表达显著上调;pDC316-SOX-9质粒转染骨髓基质细胞后可促进骨髓基质细胞的增殖,细胞有向软骨细胞分化趋势.结论 SOX9能够促进软骨形成,SOX9基因质粒转染的MSCs可望在软骨组织工程临床治疗中得到更广泛的运用.
目的 檢測併分析SOX9基因在軟骨髮育過程中的錶達規律;構建其質粒併轉染至培養的骨髓源基質細胞(BMSCs)中,通過細胞培養觀察SOX9基因對BMSCs生長特性的影響,為可能的骨關節炎(OA)基因治療提供理論基礎.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SOX9基因在軟骨髮育過程中的錶達規律;用噻唑藍(MTT)法、免疫組織化學、囌木素-伊紅(HE)染色、反轉錄-聚閤酶鏈反應(RT-PCR)及酶聯免疫吸附試驗(ELISA)法檢測SOX9基因轉染MSCs的效果及產物的錶達.結果 SOX9基因在軟骨髮育過程中的軟骨形成關鍵期錶達顯著上調;pDC316-SOX-9質粒轉染骨髓基質細胞後可促進骨髓基質細胞的增殖,細胞有嚮軟骨細胞分化趨勢.結論 SOX9能夠促進軟骨形成,SOX9基因質粒轉染的MSCs可望在軟骨組織工程臨床治療中得到更廣汎的運用.
목적 검측병분석SOX9기인재연골발육과정중적표체규률;구건기질립병전염지배양적골수원기질세포(BMSCs)중,통과세포배양관찰SOX9기인대BMSCs생장특성적영향,위가능적골관절염(OA)기인치료제공이론기출.방법 용기인심편기술건립임신태서지아연골발육과정적기인표체보,분석SOX9기인재연골발육과정중적표체규률;용새서람(MTT)법、면역조직화학、소목소-이홍(HE)염색、반전록-취합매련반응(RT-PCR)급매련면역흡부시험(ELISA)법검측SOX9기인전염MSCs적효과급산물적표체.결과 SOX9기인재연골발육과정중적연골형성관건기표체현저상조;pDC316-SOX-9질립전염골수기질세포후가촉진골수기질세포적증식,세포유향연골세포분화추세.결론 SOX9능구촉진연골형성,SOX9기인질립전염적MSCs가망재연골조직공정림상치료중득도경엄범적운용.
Objective To examine the gene expression profile of SRY-type high-mobility-group box-9(SOX9) during entochondrostosis of mice and investigate the effects of transfection of the pDC316-SOX-9 on mice mesenchymal stem cells(MSCs) in vitro. Methods cDNA microarray technique with 34 000 genes was used to analyze the gene expression profiles during entochondrostosis in the limbs of mice embryo from E10 to E14. Pathway analysis of SOX9 was performed with GCOS1.2 software. The recombined expression vector pDC316-SOX-9 was constructed and transfected into mice MSCs by lipofectamine. The phenotype changes of cells were observed with cell energometry, HE stain, immunohistochemical method, RT-PCR and ELISA.Results The gene expression of SOX9 during the critical phase of chondrogenesis in mice embryo limbs at E12 was increased evidently. SOX9 might promot chondrogenesis. As compared with vector and blank group,the chondrocytes of the SOX9 transfected group had the tendency of enhanced differentiation. Conclusion SOX9 may promote chondrogenesis. The transfection of SOX9 gene into mice MSCs can promot MSCs differentiate into chondrocyte, which may provide some experimental data for cartilage histoengineering.
