The effect of NO-donors on chloride efflux, intracellular Ca^2 + concentration and mRNA expression of CFTR and ENaC in cystic fibrosis airway epithelial cells

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• 作者：Igor Oliynyk^a ; Rashida Hussain^a ; Ahmad Amin^a ; Marie Johannesson^a ; ^b ; Godfried M. Roomans^a ; ^{godfried.roomans@oru.se}
• 关键词：Nitric oxide donors ; Cystic fibrosis ; Airway epithelium ; CFTR ; ENaC ; Calcium
• 刊名：Experimental and Molecular Pathology
• 出版年：2013
• 出版时间：June, 2013
• 年：2013
• 卷：94
• 期：3
• 页码：474-480
• 全文大小：582 K

文摘

Since previous studies showed that the endogenous bronchodilator, S-nitrosglutathione (GSNO), caused a marked increase in CFTR-mediated chloride (Cl^?) efflux and improved the trafficking of CFTR to the plasma membrane, and that also the nitric oxide (NO)-donor GEA3162 had a similar, but smaller, effect on Cl^? efflux, it was investigated whether the NO-donor properties of GSNO were relevant for its effect on Cl^? efflux from airway epithelial cells. Hence, the effect of a number of other NO-donors, sodium nitroprusside (SNP), S-nitroso-N-acetyl-dl-penicillamine (SNAP), diethylenetriamine/nitric oxide adduct (DETA-NO), and diethylenetriamine/nitric oxide adduct (DEA-NONOate) on Cl^? efflux from CFBE (?F508/?F508-CFTR) airway epithelial cells was tested. Cl^? efflux was determined using the fluorescent N-(ethoxycarbonylmethyl)-6-methoxyquinoliniu bromide (MQAE)-technique. Possible changes in the intracellular Ca^2 + concentration were tested by the fluorescent fluo-4 method in a confocal microscope system. Like previously with GSNO, after 4 h incubation with the NO-donor, an increased Cl^? efflux was found (in the order SNAP > DETA-NO > SNP). The effect of DEA-NONOate on Cl^? efflux was not significant, and the compound may have (unspecific) deleterious effects on the cells. Again, as with GSNO, after a short (5 min) incubation, SNP had no significant effect on Cl^? efflux. None of the NO-donors that had a significant effect on Cl^? efflux caused significant changes in the intracellular Ca^2 + concentration. After 4 h preincubation, SNP caused a significant increase in the mRNA expression of CFTR. SNAP and DEA-NONOate decreased the mRNA expression of all ENaC subunits significantly. DETA-NO caused a significant decrease only in ¦Á-ENaC expression. After a short preincubation, none of the NO-donors had a significant effect, neither on the expression of CFTR, nor on that of the ENaC subunits in the presence and absence of l-cysteine. It can be concluded that the effect of GSNO on Cl^? efflux is, at least in part, due to its properties as an NO-donor, and the effect is likely to be mediated by CFTR, not by Ca^2 +-activated Cl^? channels.