骨髓间充质干细胞；_Caspase3；_失巢凋亡_

大鼠骨髓间充质干细胞体外诱导失巢凋亡过程中半胱氨酸蛋白酶3的作用

http://www.100md.com 《中国矫形外科杂志》 2006年第11期

骨髓间充质干细胞；,Caspase3；,失巢凋亡,,骨髓间充质干细胞；,Caspase3；,失巢凋亡,1材料与方法,2结果

     摘要：[目的]探讨在体外诱导骨髓间充质干细胞失巢凋亡过程中半胱氨酸蛋白酶3(Caspase3)的作用。[方法]将体外培养的骨髓间充质干细胞随机分为3组：对照组、诱导凋亡组、抑制凋亡组。应用Caspase3活性检测试剂盒，荧光比色法和Western blotting法检测失巢凋亡中Caspase3活性的改变；借助流式细胞仪分析骨髓间充质干细胞凋亡的变化。[结果]流式细胞术显示诱导凋亡组出现了明显的凋亡峰，凋亡率随诱导时间的延长而明显增加；其它两组的凋亡率较低且较稳定。Western blotting检测和Caspase3活性荧光检测显示，诱导凋亡组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，以后者更灵敏，其余两组活性无明显变化。[结论]Caspase3蛋白在诱导骨髓间充质干细胞失巢凋亡的过程中发挥着重要作用；抑制Caspase3的活性可以有效降低细胞失巢凋亡率。

    关键词：骨髓间充质干细胞； Caspase3；失巢凋亡

    Significance of casepase-3 in induced anoikis of mesenchymal stem cells(MSCs)in vitro∥FENG Jianjun,YANG Shuhua,SUN Li,et alDepartment of Orthopaedics,Union Hospital,Tongji Medical College,Huazhong university of Science & Technology,Wuhan 430022

    Abstract：[Objective]To investigate the significance of Caspase3 in anoikis induced by preventing mesenchymal stem cells(MSCs)from adherence in vitro[Method]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 blotting[Result]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 blotting and fluorometric assay showed the caspase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 blotting[Conclusion]Caspase3 plays a vital role in suspended culture induced anoikis of MSCs in vitroDecreasing the activity of caspase3 by Caspase3 Inhibitor can reduce the anoikis rate ......

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