An investigation of the role of glutathione in increased epithelial permeability induced by cigarette smoke in vivo and in vitro.
Airspace epithelial permeability is known to increase in cigarette smokers. To study the role of the antioxidant reduced glutathione (GSH) in this phenomenon, we used an in vitro model of the epithelial permeability of a monolayer of human type II alveolar epithelial cells (A549 cell line). Both whole (WSC) and vapor (VSC) smoke condensates induced a recoverable, concentration-dependent increase in epithelial permeability to 125iodine-labeled bovine serum albumin (125IBSA), associated with a profound fall in intracellular GSH. Buthionine sulfoximine (BSO), a GSH synthesis inhibitor, decreased GSH levels in A549 epithelial cells, significantly increased A549 epithelial cell permeability, and enhanced both WSC and VSC-induced A549 epithelial cell permeability. Co-culturing epithelial cells and GSH (500 microM) reduced WSC-induced, but not VSC-induced A549 epithelial cell permeability. Increasing intracellular GSH also ameliorated the smoke-induced increased epithelial permeability. Concentrations of cigarette smoke condensate of < 20% increased A549 epithelial cell permeability without associated cell detachment and lysis, which was also the case with BSO-induced increased epithelial permeability. WSC and VSC, instilled intratracheally, significantly increased rat lung epithelial permeability to 125IBSA, 6 h postinstillation, associated with a significant recruitment of neutrophils into the airspaces. This was associated with a small increase in GSH in the lung tissue of VSC-treated rats. However, both WSC and VSC markedly reduced GSH in bronchoalveolar lavage (BAL) fluid. Reduction in lung GSH to 95% but not to 68% of control values by BSO increased lung epithelial permeability in vivo. However, there was no additive effect on epithelial permeability of WSC and BSO.(ABSTRACT TRUNCATED AT 250 WORDS)
Li
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Department of Medicine, City Hospital, Edinburgh, Scotland, U.K.
Donaldson
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Rahman
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MacNee
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D000818:Animals; D000963:Antimetabolites / Q000494:pharmacology; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry / Q000166:cytology; D019328:Buthionine Sulfoximine; D002417:Cattle; D002460:Cell Line; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002478:Cells, Cultured; D004848:Epithelium; D005978:Glutathione / Q000032:analysis* / Q000502:physiology*; D006801:Humans; D066298:In Vitro Techniques; D007424:Intracellular Fluid / Q000737:chemistry; D007457:Iodine Radioisotopes / Q000493:pharmacokinetics*; D007958:Leukocyte Count; D007962:Leukocytes / Q000737:chemistry; D008717:Methionine Sulfoximine / Q000031:analogs & derivatives / Q000494:pharmacology; D009504:Neutrophils / Q000502:physiology; D011650:Pulmonary Alveoli / Q000187:drug effects* / Q000473:pathology*; D051381:Rats; D017208:Rats, Wistar; D012710:Serum Albumin, Bovine / Q000493:pharmacokinetics*; D012907:Smoking / Q000009:adverse effects / Q000473:pathology*; D013997:Time Factors
D000963:Antimetabolites; D007457:Iodine Radioisotopes; D008717:Methionine Sulfoximine; D012710:Serum Albumin, Bovine; D019328:Buthionine Sulfoximine; D005978:Glutathione