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抑制钠钾ATP酶可激活人神经母细胞瘤细胞系中肿胀诱导的牛磺酸外流。

Inhibition of Na+, K+-ATPase activates swelling-induced taurine efflux in a human neuroblastoma cell line.

作者信息

Basavappa S, Mobasheri A, Errington R, Huang C C, Al-Adawi S, Ellory J C

机构信息

University Laboratory of Physiology, University of Oxford, United Kingdom.

出版信息

J Cell Physiol. 1998 Feb;174(2):145-53. doi: 10.1002/(SICI)1097-4652(199802)174:2<145::AID-JCP1>3.0.CO;2-O.

Abstract

The Na+ pump (Na+, K+-ATPase) has been implicated in the regulation of many cellular functions, including cell volume regulation. The effects of inhibiting Na+ pump activity on cell volume and taurine efflux were evaluated in the human neuroblastoma cell line CHP-100. Cell volume changes monitored with the Coulter Multisizer technique and confocal microscopy showed that neuroblastoma cells exposed to ouabain swelled by 22 +/- 4% (n = 5). The rapid cell swelling was followed by regulatory volume decrease (RVD). In cells treated with ouabain, 14C-taurine efflux increased by 183 +/- 11% compared with controls. However, cells exposed simultaneously to ouabain and hypoosmotic solution resulted in a 14C-taurine efflux of 207 +/- 18%. Western blot and immunofluorescence microscopy with specific monoclonal antibodies for the catalytic alpha isoforms of Na+, K+-ATPase demonstrated high levels of the ubiquitously expressed alpha1 and the neuronal-specific alpha3. Ouabain-binding data showed that CHP-100 cells express approximately 3 x 10(5) pump units/cell. The present data indicate that efflux of taurine may be involved during volume recovery subsequent to blockade of Na+, K+-ATPase in CHP-100 cells.

摘要

钠泵(Na⁺,K⁺ - ATP酶)与包括细胞体积调节在内的多种细胞功能的调控有关。在人神经母细胞瘤细胞系CHP - 100中评估了抑制钠泵活性对细胞体积和牛磺酸外流的影响。用库尔特多参数血细胞分析仪技术和共聚焦显微镜监测细胞体积变化,结果显示,暴露于哇巴因的神经母细胞瘤细胞肿胀了22±4%(n = 5)。快速的细胞肿胀之后是调节性容积减小(RVD)。在用哇巴因处理的细胞中,与对照相比,¹⁴C - 牛磺酸外流增加了183±11%。然而,同时暴露于哇巴因和低渗溶液的细胞,¹⁴C - 牛磺酸外流为207±18%。用针对Na⁺,K⁺ - ATP酶催化α同工型的特异性单克隆抗体进行的蛋白质印迹和免疫荧光显微镜检查显示,普遍表达的α1和神经元特异性的α3水平很高。哇巴因结合数据表明,CHP - 100细胞每个细胞表达约3×10⁵个泵单位。目前的数据表明,在CHP - 100细胞中,Na⁺,K⁺ - ATP酶被阻断后,牛磺酸外流可能参与了容积恢复过程。

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