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Journal of Bioenergetics and Biomembranes

, Volume 50, Issue 2, pp 117–129 | Cite as

Aberrant GSH reductase and NOX activities concur with defective CFTR to pro-oxidative imbalance in cystic fibrosis airways

  • L. de Bari
  • M. Favia
  • A. Bobba
  • R. Lassandro
  • L. Guerra
  • A. Atlante
Article

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.

Keywords

Cystic fibrosis Mitochondria NADPH oxidase Glutathione reductase Cysteine 

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

Notes

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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Istituto di Biomembrane, Bioenergetica e Biotecnologie Molecolari (IBIOM) – CNRBariItaly
  2. 2.Dipartimento di BioscienzeBiotecnologie e Biofarmaceutica - Università di BariBariItaly
  3. 3.Istituto di Cristallografia (IC) – CNRBariItaly

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