Cigarette smoke extract inhibits expression of peroxiredoxin V and increases airway epithelial permeability.
Inhaled cigarette smoke induces oxidative stress in the epithelium of airways. Peroxiredoxin V (PRXV) is a potent antioxidant protein, highly expressed in cells of the airway epithelium. The goal of our study was to determine whether cigarette smoke extract (CSE) influenced expression of this protein in airway epithelia in vivo and in vitro. In Sprague-Dawley rats, we determined effects of CSE on airway epithelial permeability, mRNA levels and expression of PRXV protein. Exposure of isolated tracheal segment in vitro to 20% CSE for 4 h resulted in development of increased permeability to albumin, significantly reduced mRNA levels for PRXV, and reduced amounts of PRXV protein in the epithelium. In cultures of the airway epithelial cell lines (Calu-3, JME), primary airway cell culture (cow), and alveolar epithelial cells A549, CSE also significantly decreased transepithelial electrical resistance and expression of PRXV protein, and induced glutathione and protein oxidation. To demonstrate functional importance of PRXV, we exposed clones of HeLa cells with siRNA-downregulated PRXV to hydrogen peroxide, which resulted in increased rate of cell death and protein oxidation. CSE directly downregulates expression of functionally important antioxidant enzyme PRXV in the epithelial cells of airways, which represents one pathophysiological mechanism of cigarette smoke toxicity.
Serikov
Vladimir B
VB
Children's Hospital Oakland Research Institute, Oakland, California 94609, USA. vserikov@chori.org
Leutenegger
Christian
C
Krutilina
Raisa
R
Kropotov
Andrei
A
Pleskach
Nadezhda
N
Suh
Jung H
JH
Tomilin
Nikolay V
NV
D000818:Animals; D017209:Apoptosis; D017097:Electric Impedance; D004847:Epithelial Cells / Q000378:metabolism; D005786:Gene Expression Regulation; D008297:Male; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D018384:Oxidative Stress; D010539:Permeability; D010544:Peroxidases / Q000235:genetics*; D054464:Peroxiredoxins; D011506:Proteins / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D012906:Smoke / Q000009:adverse effects*; D014026:Nicotiana / Q000009:adverse effects*; D014132:Trachea / Q000187:drug effects* / Q000378:metabolism
D011506:Proteins; D012333:RNA, Messenger; D012906:Smoke; D009240:N-Formylmethionine Leucyl-Phenylalanine; D010544:Peroxidases; D054464:Peroxiredoxins