哇巴因诱导Jurkat细胞凋亡与胱冬酶-3及bcl-2基因家族的关系

Ouabain-induced Apoptosis of Jurkat Cells Correlates with Acti-vation of caspase-3 and Regulation of bcl-2 Gene Family

本研究探讨哇巴因诱导Jurkat细胞凋亡及其机制。采用四甲基偶氮唑盐(MTT)法观察哇巴因对Jurkat细胞生长的抑制作用,应用流式细胞术、原位末端标记技术(TUNEL)等观察细胞凋亡,应用Western-blot测定胱冬酶-3(caspase-3)的活性亚单位及Bcl-2、Bax蛋白表达水平,用比色法检测caspase-3活性。结果表明:哇巴因能诱导Jurkat细胞凋亡,在细胞凋亡过程中,Bax蛋白表达增加,caspase-3活性明显升高。结论:哇巴因可能通过调节Bcl-2、Bax蛋白表达,从而激活caspase途径诱导Jurkat细胞凋亡。

The study was aimed to explore the apoptosis effect of ouabain on Jurkat cells and its mechanism. MTT method was used to observe the inhibitory effect of ouabain on Jurkat cell proliferation. Apoptosis was detected by using flow cytometry （FCM） and the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick endlabeling reaction method （TUNEL）. The protein expressions of Bax, Bcl-2 and active subunits of caspase-3 were measured by Western blot. Activities of caspase-3 were determined by colorimetry method. The results showed that ouabain could induce apoptosis of Jurkat cells, the expression of Bax protein in process of cell apoptosis, caspase-3 activity of Jurkat cells were remarkably enhanced after ouabain treatment. It is concluded that ouabain may induce apoptosis of Jurkat cells due to the activation of caspase-3 resulting from regulation of Bax protein and bcl-2 gene expressions.