PDF
ePub
Citation
Print
An-Soo Jang, M.D., Jong-Sook Park, M.D., June-Hyuk Lee, M.D., Sung-Woo Park, M.D., Do-Jin Kim, M.D., Soo-Tak Uh, M.D., Young-Hoon Kim, M.D., Choon-Sik Park, M.D.
Abstract
Background
Aquaporins (AQPs) may play a role in the pathogenesis of pulmonary inflammation and edema. This study investigated the role ofAQPs in acute lung injury following bleomycin inhalation in rats.
Methods
Sprague-Dawley rats were treated via inhalation with 10 U/kg bleomycin hydrochloride dissolved in 5 ml of normal saline. The control rats were treated with 5 ml normal saline. The animals (n = 6-8 rats per group) were sacrificed at 4, 7, and 14 d. The changes in AQP1, AQP4, and AQP5 expression levels over time were analyzed by Western blotting. The nitrate and nitrite concentrations in the bronchoalveolar lavage fluid (BALF) were measured using a modified Griess reaction. ELISA was used to check cytokines.
Results
The respiration rates were significantly higher 4 and 7 days after the bleomycin treatment compared with those of the control rats. The tidal volume was lower in rats at 4 days after the bleomycin treatment, and the wet/dry weights of the lung were significantly higher than those of the control group. The nitrite and nitrate concentrations in the BALF from the rats at 4 days after exposure to bleomycin were greater than those from the saline-treated rats. Immunoblotting studies demonstrated that the AQP1 and AQP4 expression levels were lower in the rats at 4 days. However, the AQP4 expression level was higher at 7 days. The AQP5 expression level increased at 4, 7 and 14 days after the bleomycin treatment.
Figures and Tables
Figure 1
Increased lung fluid after bleomycin treatment. *p<0.05 compared with saline treatment, †p<0.01 compared with 4 days after bleomycin treatment.
Figure 2
Pulmonary infiltrates were observed in the peribronchial and perivascular regions with H& E stains. Original magnification, ×100. Alveolar interstitial edema was detected as thickened alveolar septa in some regions. Evidence of pulmonary edema was generally more evident in the lungs of rats at 4 days after bleomycin treatment (B) than in rats treated with saline alone (A).
References
1. Doyle RL, Szaflarski N, Modin GW, Wiener-Kronish JP, Matthay MA. Identification of patients with acute lung injury: predictors of mortality. Am J Respir Crit Care Med. 1995. 152:1818–1824.
2. Jules-Elysee K, White DA. Bleomycin-induced pulm onary toxicity. Clin Chest Med. 1990. 11:1–20.
3. Piguet PF, Vesin C, Grau GE, Thompson RC. Inte rleukin 1 receptor antagonist (IL-1ra) prevents or cures pulmonary fibrosis elicited in mice by bleomycin or silica. Cytokine. 1993. 5:57–61.
4. Smith RE, Strieter RM, Zhang K, Phan SH, Standiford TJ, Lukacs NW, et al. A role for C-C chemokines in fibrotic lung disease. J Leukoc Biol. 1995. 57:782–787.
5. Piguet PF, Collart MA, Grau GE, Kapanci Y, Vassalli P. Tumor necrosis factor/cachectin plays a key role in bleomycin-induced pneumopathy and fibrosis. J Exp Med. 1989. 170:655–663.
6. King LS, Nielsen S, Agre P. Aquaporin-1 water channel protein in lung: ontogeny, steroid-induced expression, and distribution in rat. J Clin Invest. 1996. 97:2183–2191.
7. Nielsen S, King LS, Christensen BM, Arge P. Aquaporins in complex tissues: II. cellular and subcellular distribution in respiratory tract and glands of rat. Am J Physiol. 1997. 273:C1549–C1561.
8. Towne JE, Harrod KS, Krane CM, Menon AG. Decreased expression of aquaporin (AQP) 1 and AQP5 in mouse lung after acute viral infection. Am J Respir Cell Mol Biol. 2000. 22:34–44.
9. Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, et al. Measurement of protein using bicinchninic acid. Anal Biochem. 1985. 150:76–85.
10. Dickason RR, Huston MM, Huston DP. Enhanced detection of human interluekin-5 in biological fluids utilizing murine monoclonal antibodies which delineate didtinct neutralizing epitope. Cytokine. 1994. 6:647–656.
11. Matthay MA, Zimmerman GA. Acute lung injury and the acute respiratory distress syndrome: four decades of inquiry into pathogenesis and rational management. Am J Respir Cell Mol Biol. 2005. 33:319–327.
12. Ashbaugh DG, Bigelow DB, Petty TL, Levine BE. Acute respiratory distress in adults. Lancet. 1967. 2:319. 323.
13. The Acute Respiratory Distress Syndrome Network. Ventilation with lower tidal volumes as compared with traditional tidal volumes for acute lung injury and the acute respiratory distress syndrome. N Engl J Med. 2000. 342:1301–1308.
14. Brower RG, Lanken PN, MacIntyre N, Matthay MA, Morris A, Ancukiewicz M, et al. Higher versus lower positive end-expiratory pressures in patients with the acute respiratory distress syndrome. N Engl J Med. 2004. 351:327. 336.
15. Zimmerman GA, Albertine KH, McIntyre TM. Lenfant C, Matthay MA, editors. Pathogenesis of sepsis and septic-induced lung inury. Acute respiratory distress syndrome. 2003. 1st ed. New York: Marcel Dekker;245–287.
16. Jang AS, Lee JU, Choi IS, Park KO, Lee JH, Park SW, et al. Expression of nitric oxide synthase, aquaporin 1 and aquaporin 5 in rat after bleomycin inhalation. Intensive Care Med. 2004. 30:489–495.
17. Moncada S, Palmer RM, Higgs EA. Biosynthesis of nitric oxide from L-arginine: a pathway for the regulation of cell function and communication. Biochem Pharmacol. 1989. 38:1709–1715.
18. Moncada S, Palmer RMJ, Higgs EA. Nitric oxide: physiology, pathophysiology and pharmacology. Pharmacol Rev. 1991. 43:109–142.
19. Haddad IY, Pataki G, Hu P, Galliani C, Beckman JS, Matalon S. Quantification of nitrotyrosine levels in lung secretions of patients and animals with acute lung injury. J Clin Invest. 1994. 94:2407–2413.
20. Agre P, Saboori AM, Asimos A, Smith BL. Purification and partial characterization of the Mr 30,000 integral membrane protein associated with the erythrocyte Rh(D) antigen. J Biol Chem. 1987. 262:17497–17503.
21. Denker BM, Smith BL, Kuhajda FP, Agre P. Identification, purification, and partial characterization of a novel Mr 28,000 integral membrane protein from erythrocytes and renal tubules. J Biol Chem. 1988. 263:15634–15642.
22. Preston GM, Agre P. Isolation of the cDNA for erythrocyte integral membrane protein of 28 kilodaltons: member of an ancient channel family. Proc Natl Acad Sci U S A. 1991. 88:11110–11114.
23. Preston GM, Carroll TP, Guggino WB, Agre P. Appearance of water channels in Xenopus oocytes expressing red cell CHIP28 protein. Science. 1992. 256:385–387.
24. Jung JS, Preston GM, Smith BL, Guggino WB, Agre P. Molecular structure of the water channel through aquaporin CHIP. J Biol Chem. 1994. 269:14648–14654.
25. van Hoek AN, Yang B, Kimiz S, Brown D. Freez-fracture analysis of plasma membrane of CHO cells stably expressing aquaporin1-5. J Membr Biol. 1998. 165:243–254.
26. Yang B, Brown D, Verkman AS. The mercurial insensitive water channel (AQP4) forms orthogonal arrays in stably transfected Chinese hamster ovary cells. J Biol Chem. 1996. 271:4577–4580.
XML Download