Organelle redox of CF and CFTR-corrected airway epithelia.
Organelle redox of CF and CFTR-corrected airway epithelia.
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CF 和 CFTR 校正的气道上皮细胞的细胞器氧化还原。
DOI：
10.1016/j.freeradbiomed.2007.04.015
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发表时间：
2007
影响因子：
7.4
通讯作者：
Machen,TerryE
中科院分区：
文献类型：
--
作者：
Schwarzer,Christian;Illek,Beate;Suh,JungH;Remington,SJames;Fischer,Horst;Machen,TerryE
在囊性纤维化中，CFTR 功能降低可能会改变气道上皮细胞的氧化还原特性。使用氧化还原敏感的 GFP (roGFP1) 和成像显微镜来测量囊性纤维化鼻上皮细胞和 CFTR 校正细胞的细胞质、内质网 (ER)、线粒体和细胞表面的氧化还原电位。我们还测量了细胞裂解物和顶端液中的谷胱甘肽和半胱氨酸硫醇氧化还原状态，以覆盖可能受 CFTR 影响的一系列氧化还原电位和环境。使用 roGFP1 测量，CF 和 CFTR 校正细胞的细胞表面氧化还原电位（约 -207±8 mV）和 ER（约 -217±1 mV）以及 DTT 处理后顶端液和 ER 腔的调节速率相似。 CF 和 CFTR 校正细胞在线粒体 (−344±9 mV) 和细胞质 (−322±7 mV) 中具有相似的氧化还原电位。 CF 和 CFTR 校正细胞中羧基二氯二氢荧光素二乙酸酯和顶端 Amplex 红的氧化发生率相同。 CF 和 CFTR 校正细胞中细胞裂解物和顶端液中的谷胱甘肽和半胱氨酸氧化还原对相等。这些细胞器氧化还原电位的定量估计与使用小分子对的顶端和细胞测量相结合，证实了 CF 和 CFTR 校正细胞的氧化还原特性没有差异。
In cystic fibrosis reduced CFTR function may alter redox properties of airway epithelial cells. Redox-sensitive GFP (roGFP1) and imaging microscopy were used to measure the redox potentials of the cytosol, endoplasmic reticulum (ER), mitochondria, and cell surface of cystic fibrosis nasal epithelial cells and CFTR-corrected cells. We also measured glutathione and cysteine thiol redox states in cell lysates and apical fluids to provide coverage over a range of redox potentials and environments that might be affected by CFTR. As measured with roGFP1, redox potentials at the cell surface (approx −207±8 mV) and in the ER (approx −217±1 mV) and rates of regulation of the apical fluid and ER lumen after DTT treatment were similar for CF and CFTR-corrected cells. CF and CFTR-corrected cells had similar redox potentials in mitochondria (−344±9 mV) and cytosol (−322±7 mV). Oxidation of carboxydichlorodihydrofluorescein diacetate and of apical Amplex red occurred at equal rates in CF and CFTR-corrected cells. Glutathione and cysteine redox couples in cell lysates and apical fluid were equal in CF and CFTR-corrected cells. These quantitative estimates of organelle redox potentials combined with apical and cell measurements using small-molecule couples confirmed there were no differences in the redox properties of CF and CFTR-corrected cells.