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Thiol-redox antioxidants protect against lung vascular endothelial cytoskeletal alterations caused by pulmonary fibrosis inducer, bleomycin: comparison between classical thiol-protectant, N-acetyl-L-cysteine, and novel thiol antioxidant, N,N'-bis-2-mercaptoethyl isophthalamide.
- Source :
-
Toxicology mechanisms and methods [Toxicol Mech Methods] 2012 Jun; Vol. 22 (5), pp. 383-96. - Publication Year :
- 2012
-
Abstract
- Lung vascular alterations and pulmonary hypertension associated with oxidative stress have been reported to be involved in idiopathic lung fibrosis (ILF). Therefore, here, we hypothesize that the widely used lung fibrosis inducer, bleomycin, would cause cytoskeletal rearrangement through thiol-redox alterations in the cultured lung vascular endothelial cell (EC) monolayers. We exposed the monolayers of primary bovine pulmonary artery ECs to bleomycin (10 µg) and studied the cytotoxicity, cytoskeletal rearrangements, and the macromolecule (fluorescein isothiocyanate-dextran, 70,000 mol. wt.) paracellular transport in the absence and presence of two thiol-redox protectants, the classic water-soluble N-acetyl-L-cysteine (NAC) and the novel hydrophobic N,N'-bis-2-mercaptoethyl isophthalamide (NBMI). Our results revealed that bleomycin induced cytotoxicity (lactate dehydrogenase leak), morphological alterations (rounding of cells and filipodia formation), and cytoskeletal rearrangement (actin stress fiber formation and alterations of tight junction proteins, ZO-1 and occludin) in a dose-dependent fashion. Furthermore, our study demonstrated the formation of reactive oxygen species, loss of thiols (glutathione, GSH), EC barrier dysfunction (decrease of transendothelial electrical resistance), and enhanced paracellular transport (leak) of macromolecules. The observed bleomycin-induced EC alterations were attenuated by both NAC and NBMI, revealing that the novel hydrophobic thiol-protectant, NBMI, was more effective at µM concentrations as compared to the water-soluble NAC that was effective at mM concentrations in offering protection against the bleomycin-induced EC alterations. Overall, the results of the current study suggested the central role of thiol-redox in vascular EC dysfunction associated with ILF.
- Subjects :
- Acetylcysteine chemistry
Actin Cytoskeleton metabolism
Actin Cytoskeleton pathology
Animals
Antioxidants chemistry
Cattle
Cell Culture Techniques
Cell Survival drug effects
Cells, Cultured
Cysteamine chemistry
Cysteamine pharmacology
Endothelial Cells drug effects
Endothelial Cells metabolism
Endothelial Cells pathology
Endothelium, Vascular metabolism
Endothelium, Vascular pathology
Glutathione metabolism
Idiopathic Pulmonary Fibrosis metabolism
Idiopathic Pulmonary Fibrosis pathology
Lung blood supply
Lung metabolism
Lung pathology
Microscopy, Fluorescence
Molecular Structure
Oxidation-Reduction
Phthalic Acids chemistry
Reactive Oxygen Species metabolism
Structure-Activity Relationship
Sulfhydryl Compounds chemistry
Acetylcysteine pharmacology
Actin Cytoskeleton drug effects
Antioxidants pharmacology
Bleomycin pharmacology
Cysteamine analogs & derivatives
Endothelium, Vascular drug effects
Idiopathic Pulmonary Fibrosis prevention & control
Lung drug effects
Phthalic Acids pharmacology
Sulfhydryl Compounds pharmacology
Subjects
Details
- Language :
- English
- ISSN :
- 1537-6524
- Volume :
- 22
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Toxicology mechanisms and methods
- Publication Type :
- Academic Journal
- Accession number :
- 22409285
- Full Text :
- https://doi.org/10.3109/15376516.2012.673089