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目的：观察低浓度哇巴因对人白血病细胞株Jurkat 生长的影响并初步探讨哇巴因特异性调节Jurkat 细胞生长的机制。方法：分别用不同低浓度哇巴因作用于人白血病细胞株Jurkat 后，采用四甲基偶氮唑盐MTT 法检测细胞增殖情况、流式细胞学FCM 技术检测细胞凋亡情况以及细胞内线粒体膜电位变化情况，Western blot 法观察哇巴因对Jutkat 细胞膜表面钠钾ATP 酶的表达调节作用。结果：MTT 及FCM 检测结果表明随着哇巴因浓度的增高, 哇巴因对Jurkat 细胞的增殖抑制及促凋亡作用越明显。 WesternBlot 结果显示30nM 及50nM 哇巴因作用于Jurkat 细胞株48h 后引起细胞钠钾ATP 酶表达下调，[3H]- 哇巴因结合实验结果显示在Jurkat 细胞株随着哇巴因作用浓度升高，细胞膜钠泵对哇巴因的亲和力逐渐下降。结论：低浓度哇巴因即可抑制白血病细胞株Jurkat 增殖并诱导其凋亡。这种特异性细胞生长调控作用与哇巴因引起的细胞膜钠钾ATP 酶表达变化相关，最终引起细胞内线粒体膜电位发生变化，释放相关凋亡蛋白,诱导细胞凋亡。

英文摘要:

Objective: To investigate the effect and the mechanism of ouabain at low concentrations on the growth in the Jurkat leukemia cell lines. Methods: Cell proliferation was observed by using MTT method.The cell apoptosis and the mitochondrial membrane potential changes in cells were assayed by Streaming cytology technology. The expressions of Na/K-ATPase α1 Subunit were evaluated by western-blot before and after the cells were treated with different low concentration ouabain. Results: MTT tests showed that with the increasing of the concentration of ouabain, the proliferation of the Jurkat cell suppressed and the apoptosis role increasing. The result of western-blot showed that low concentrations of ouabain (30nM 50nM) caused Na/K-ATPase expression fall on the Jurkat cell for 48h. The [3H]-ouabain combining tests showed that with the increasing of the concentration,of ouabain the active Na/K-ATPase in the plasma membrane gradually decreased. Conclusions: The low concentration ouabain can not only inhibit the proliferation of the Jurkat cells, but also induce its apoptosis. This effect and the Na/K-ATPase expression changes related. That changes caused the mitochondrial membrane potential to change finally thus to induce the apoptosis by releasing the apoptosis proteins.

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