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首页> 外文期刊>Journal of Huazhong University of Science and Technology >Role of Caspase-3 Inhibitor in Induced Anoikis of Mesenchymal Stem Cells In Vitro
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Role of Caspase-3 Inhibitor in Induced Anoikis of Mesenchymal Stem Cells In Vitro

机译：Caspase-3抑制剂在诱导间充质干细胞失语症中的作用

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By preventing mesenchymal stem cells (MSCs) from adhering to precoated agarose to create a model of MSC suspension in vitro, we investigated anoikis in MSCs and the role of caspase-3 in the anoikis. The cultured MSCs were randomly divided into 3 groups: the anoikis group, caspase-3 inhibitor group and control group. Before experiment, we coated dishes with 1.5 % agarose; in the anoikis group, MSCs were put into the precoated dishes; and in the inhibitor group, caspase-3 inhibitor and MSCs were also put into the precoated dishes; but there were not intervention in the control group. MSCs were collected at 2 h, 6 h, 12 h and 24 h. The alteration of caspase-3 activity was evaluated by caspase-3 fluorometric assay and western blot analysis. The apoptosis rates were detected by flow cytometry. MSCs were round and suspended sufficiently in the anoikis and inhibitor groups. Caspase-3 fluorometric assay showed that there were significant differences in statistics between the anoikis group and the others (P < 0.05). Western blot analysis discovered that caspase-3 expression in the anoikis group was more than that in the control and inhibitor groups (P < 0.05). Flow cytometry showed that the apoptosis peak appeared in all the three groups, but it increased dramatically in the anoikis group. The apoptosis rates in the inhibitor and control groups were low and stable. And there were significant differences in statistics between the anoikis group and the others (P < 0.05). MSCs will undergo anoikis in suspended condition if they are separated from the extracellular matrix. Caspase-3 inhibitors can suppress caspase-3 activity and reduce the apoptosis rate significantly. Caspase-3 plays a vital part in induced MSC anoikis in vitro. MSCs suspension culture system might be set up with argorose and caspase-3 inhibitor.
机译：通过防止间充质干细胞（MSCs）粘附到预先包被的琼脂糖上以建立体外MSC悬浮液模型，我们研究了MSCs中的anoikis和caspase-3在anoikis中的作用。将培养的MSCs随机分为3组：无神经组，caspase-3抑制剂组和对照组。在实验之前，我们用1.5％的琼脂糖包被菜。在anoikis组中，将MSCs放入预涂层的盘子中。在抑制剂组中，将caspase-3抑制剂和MSCs也放入预涂盘中。但对照组没有干预。在2小时，6小时，12小时和24小时收集MSC。 caspase-3活性的变化通过caspase-3荧光测定和蛋白质印迹分析进行评估。用流式细胞仪检测细胞凋亡率。 MSC是圆形的，并充分悬浮于失神经和抑制剂组中。 Caspase-3荧光测定法显示，无神经症组与其他组之间的统计学差异有统计学意义（P <0.05）。蛋白质印迹分析发现，anoikis组的caspase-3表达高于对照组和抑制剂组（P <0.05）。流式细胞仪检测发现，三组均出现凋亡高峰，而无神经组则明显升高。抑制剂和对照组的细胞凋亡率低且稳定。 anoikis组与其他组之间的统计学差异有统计学意义（P <0.05）。如果将MSC与细胞外基质分开，它们将处于悬浮状态。 Caspase-3抑制剂可抑制caspase-3活性并显着降低细胞凋亡率。 Caspase-3在体外诱导的MSC阳极中起着至关重要的作用。 MSCs悬浮培养系统可能建立有阿果糖和caspase-3抑制剂。

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