CAJ | 학술논문

目的观察Caspase-3在小鼠胚胎肢芽间质干细胞分化过程中的作用,并将其运用于体外间质干细胞模型中进行验证.方法用基因芯片技术检测Caspase-3在小鼠胚胎肢芽间质干细胞分化过程中的基因表达,分析其表达规律和可能作用;Yg用Caspase-3活性检测试剂盒,荧光比色法和Western blot法检测体外培养的骨髓间质干细胞诱导失巢凋亡中Caspme-3活性的改变;借助流式细胞仪分析骨髓间质干细胞凋亡的变化.结果 Caspase-3在小鼠胚胎肢芽间质干细胞向软骨分化的关键期(E12)出现显著的表达下调;Caspase-3的活性以及蛋白表达水平随诱导凋亡时间的延长而明显升高,且与细胞凋亡率相关.结论 Caspase-3蛋白在体内和体外诱导间质干细胞凋亡的过程中均发挥着重要作用;胚胎发育过程中,通过下调Caspase-3的表达以促进软骨形成,而抑制Caspase-3的活性可以有效降低细胞凋亡率.
목적 관찰Caspase-3재소서배태지아간질간세포분화과정중적작용,병장기운용우체외간질간세포모형중진행험증.방법 용기인심편기술검측Caspase-3재소서배태지아간질간세포분화과정중적기인표체,분석기표체규률화가능작용;Yg용Caspase-3활성검측시제합,형광비색법화Western blot법검측체외배양적골수간질간세포유도실소조망중Caspme-3활성적개변;차조류식세포의분석골수간질간세포조망적변화.결과 Caspase-3재소서배태지아간질간세포향연골분화적관건기(E12)출현현저적표체하조;Caspase-3적활성이급단백표체수평수유도조망시간적연장이명현승고,차여세포조망솔상관.결론 Caspase-3단백재체내화체외유도간질간세포조망적과정중균발휘착중요작용;배태발육과정중,통과하조Caspase-3적표체이촉진연골형성,이억제Caspase-3적활성가이유효강저세포조망솔.
Objective To examine the gene expression profile of Casepase-3 during entochondrostosis of mice and explore the expression rules and effects of it;investigate the significance of Caspase-3 in anoikis induced by preventing mesenchymal stem cells (MSCs)from adherence in vitr. 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 EIO to El4. Pathway analysis of Casepase-3 was performed with GCOS1.2 software. The cultured MSCs were randomly divided into 3 groups:control group,induced group and Caspase-3 inhibitor group. MSCs anoikis was detected by flow cytometry. The Caspase-3 activity was evaluated by Caspase-3 Fluorometric Assay Kit and Western blot. Results There were obvious down regulation of Caspase-3 during the critical period of entochondrostosis(E12) in microarray. The flow cytometry analysis showd that apoptosis peak was appeared significantly in the induced groupand increased dramaticly along with the apoptosis conditions going on. The apoptosis rate of the others were lower and stable. Western blot and fluorometric assay showed the caspas e-3 expressive level or activity in induced group Was the highest and Was related with its apoptosis rate. The fluorometric assay technique Was sensitive to the western blot. Conclusion This finding revealed Caspase-3 plays a vital role in apoptosis of MSCs in vivo and in vitro. Down regulation of Caspase-3 during the critical period of entochondrostosis(El2) promote chondrogenesis in mice embryonal limb bud. Decreasing the activity of Caspase-3 by Caspase-3 Inhibitor can reduce the anoikis rat in vitro.