Abstract
Cystic fibrosis (CF) is associated to impaired Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) channel also causing decreased glutathione (GSH) secretion, defective airway bacterial clearance and inflammation. Here we checked the main ROS-producing and ROS-scavenging enzymes as potential additional factors involved in CF pathogenesis. We found that CFBE41o-cells, expressing F508del CFTR, have increased NADPH oxidase (NOX) activity and expression level, mainly responsible of the increased ROS production, and decreased glutathione reductase (GR) activity, not dependent on GR protein level decrease. Furthermore, defective CFTR proved to cause both extracellular and intracellular GSH level decrease, probably by reducing the amount of extracellular GSH-derived cysteine required for cytosolic GSH synthesis. Importantly, we provide evidence that defective CFTR and NOX/GR activity imbalance both contribute to NADPH and GSH level decrease and ROS overproduction in CF cells.
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Abbreviations
- ACI:
-
Acivicin
- ADH:
-
Alcohol dehydrogenase
- ALLO:
-
Allopurinol
- AOX:
-
Antioxidant system
- ASL:
-
Airway surface liquid
- CF:
-
Cystic Fibrosis
- CFBE:
-
CFBE41o-cells expressing F508del CFTR
- CFTR:
-
Cystic Fibrosis Transmembrane Conductance Regulator
- COX:
-
Mitochondrial Complex IV
- CP:
-
Captopril
- CYS:
-
Cysteine
- CYTP450:
-
Cytochrome P450
- DMSO:
-
Dimethyl sulfoxide
- DPI:
-
Diphenyliodonium
- DTNB:
-
5,5′-dithio-bis(2-nitrobenzoic acid)
- DUOX 1:
-
Dual oxidase 1
- DUOX 2:
-
Dual oxidase 2
- exGSH:
-
Extracellular GSH
- GPX:
-
Glutathione peroxidase
- GR:
-
Glutathione reductase
- GSH:
-
Reduced glutathione
- GSSG:
-
Glutathione disulfide
- γGT:
-
Γ-glutamyltransferase
- H2O2 :
-
Hydrogen peroxide
- inGSH:
-
Intracellular GSH
- L-NAME:
-
N(ω)-nitro-L-arginine methyl ester
- mtCx-I:
-
mitochondrial Complex I
- MP:
-
Metyrapon
- MTT:
-
3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide
- NH2TZ:
-
NH2-triazole
- NOX:
-
NAD(P)H oxidases
- O2 −• :
-
Superoxide anion radical
- PBS:
-
Phosphate-buffered saline
- PES:
-
Phenazine ethosulfate
- ROS:
-
Reactive oxygen species
- ROT:
-
Rotenone
- S.D.:
-
Standard deviation
- TNB:
-
5′-thio-2-nitrobenzoic acid
- Wt-CFBE:
-
CFBE41o-cells stably expressing wildtype CFTR
- XOD:
-
Xanthine oxidase
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Acknowledgments
This research was supported by Italian Cystic Fibrosis Research Foundation with the contribution of “Infront e Play for Change” and “Gare di golf” (FFC#1/2015 Project: “Relationship between mitochondria and F508del-CFTR in Cystic Fibrosis”) to A.A.
M.F. has been PostDoc fellow of the Italian Cystic Fibrosis Research Foundation.
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L. Guerra and A. Atlante share the last authorship.
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de Bari, L., Favia, M., Bobba, A. et al. Aberrant GSH reductase and NOX activities concur with defective CFTR to pro-oxidative imbalance in cystic fibrosis airways. J Bioenerg Biomembr 50, 117–129 (2018). https://doi.org/10.1007/s10863-018-9748-x
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DOI: https://doi.org/10.1007/s10863-018-9748-x