1. Short Term Exposure of Sheep Tracheal Epithelium to Cigarette Smoke Extract Reduces ENaC Current: A Pilot Study.
- Author
-
Jagirdar RM, Grammatikopoulos A, Ioannou M, Solenov E, Gourgoulianis KI, Hatzoglou C, Giannou AD, Mercanoglu B, and Zarogiannis SG
- Subjects
- Animals, Sheep, Pilot Projects, Smoke adverse effects, Amiloride pharmacology, Respiratory Mucosa metabolism, Respiratory Mucosa drug effects, Respiratory Mucosa pathology, Epithelium drug effects, Epithelium metabolism, Epithelium pathology, Epithelial Sodium Channels metabolism, Trachea metabolism, Trachea drug effects, Trachea pathology
- Abstract
Background/aim: Cigarette smoke has been shown to induce a phenotype in humans known as "acquired cystic fibrosis". This occurs because the cystic fibrosis transmembrane conductance regulator (CFTR) functions are impaired systemically due to the deleterious effects of smoke components. Elucidation of cigarette smoke effects on the tracheal epithelium is important. The aim of this study was to develop an ex vivo sheep tracheal model to investigate tracheal ion function. In this model, the epithelial sodium channel (ENaC) is inhibited after exposure to cigarette smoke extract (CSE) as a proof of principle., Materials and Methods: Tracheas were isolated from healthy sheep and the tracheal epithelium was surgically excised. Tissues were mounted in Ussing chambers and the short circuit current (I
sc ) was measured after incubation with 5% CSE in PBS or PBS alone for 30 min. The function of ENaC was investigated by the addition of amiloride (10-5 M) apically. Western blot analysis was performed to assess differences in ENaC quantity after CSE exposure. Some specimens were stained with H&E for detection of histological alterations., Results: The amiloride effect on normal epithelium led to a significant decrease in Isc [ΔI=33±5.92 μA/cm2 ; p<0.001 versus control experiments (ΔI=1.44±0.71 μA/cm2 )]. After incubation with CSE, ENaC Isc was significantly reduced (ΔI=14.80±1.96 μA/cm2 ; p<0.001). No differences in αENaC expression were observed between CSE-exposed and normal tracheal epithelium. Histological images post CSE incubation revealed decreases in the height of the epithelium, with basal cell hyperplasia and loss of ciliated cells., Conclusion: Reduced ENaC inhibition by amiloride after CSE incubation could be due to alterations in the tracheal epithelium., (Copyright © 2024, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)- Published
- 2024
- Full Text
- View/download PDF