Tuberculosis and Respiratory Diseases

The Korean Academy of Tuberculosis and Respiratory Diseases (대한결핵및호흡기학회)
권호 표지

Aquaporin in bleomycin induced lung injury

급성 폐손상 동물모델에서 aquaporin 수분통로의 발현

  • Jang, An-Soo (Division of Allergy and Respintoy Diseases, Depotmet of Intenal Medicine Soonchunhyang University Hospital) ;
  • Park, Jong-Sook (Division of Allergy and Respintoy Diseases, Depotmet of Intenal Medicine Soonchunhyang University Hospital) ;
  • Lee, June-Hyuk (Division of Allergy and Respintoy Diseases, Depotmet of Intenal Medicine Soonchunhyang University Hospital) ;
  • Park, Sung-Woo (Division of Allergy and Respintoy Diseases, Depotmet of Intenal Medicine Soonchunhyang University Hospital) ;
  • Kim, Do-Jin (Division of Allergy and Respintoy Diseases, Depotmet of Intenal Medicine Soonchunhyang University Hospital) ;
  • Uh, Soo-Taek (Division of Allergy and Respintoy Diseases, Depotmet of Intenal Medicine Soonchunhyang University Hospital) ;
  • Kim, Yong-Hoon (Division of Allergy and Respintoy Diseases, Depotmet of Intenal Medicine Soonchunhyang University Hospital) ;
  • Park, Choon-Sik (Division of Allergy and Respintoy Diseases, Depotmet of Intenal Medicine Soonchunhyang University Hospital)
  • 장안수 (순천향대학교 의과대학 내과학교실) ;
  • 박종숙 (순천향대학교 의과대학 내과학교실) ;
  • 이준혁 (순천향대학교 의과대학 내과학교실) ;
  • 박성우 (순천향대학교 의과대학 내과학교실) ;
  • 김도진 (순천향대학교 의과대학 내과학교실) ;
  • 어수택 (순천향대학교 의과대학 내과학교실) ;
  • 김용훈 (순천향대학교 의과대학 내과학교실) ;
  • 박춘식 (순천향대학교 의과대학 내과학교실)
  • Received : 2005.12.14
  • Accepted : 2006.02.28
  • Published : 2006.03.30
Download PDF
⟨ Previous Next ⟩

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. Conclusion : This study demonstrates that AQPs are expressed differently in bleomycin-induced pulmonary edema.

배 경: 급성폐손상은 여러 가지 치료의 발전에도 높은 사망률을 보이는 질환으로 병태생리학적 연구가 필요한 실정이다. 아쿠아포린 (AQP)은 수분통로로 급성폐손상에서 폐부종에 관여되는 것으로 알려져 있어 bleomycin을 이용한 급성 폐손상모델에서 아쿠아포린의 발현을 확인하고자 하였다. 방 법: Bleomycin 10 U/kg을 Sprague-Dawley rats에 흡입시켜 급성 폐손상 보델을 만들고 흡입후 4, 7, 14일에 호흡수, 상시호흡량을 측정하고, 기관지폐포세척액을 얻으며, 폐조직을 얻었다. 얻은 폐조직에서 wet/dry 량을 측정하고 기관지 폐포세척액에서 사이토카인, 산화질소산물(nitrite와 nitrate)등을 측정하였다. 그리고 폐조직에서 AQP1, AQP4와 AQP5를 면역조직염색에서 확인하였다. 결 과: Bleomycin 흡입 후 호흡수는 4, 7일에 상시호흡량은 4일에 대조군에 비해 유의한 차이를 보였다. Wet/dry 질량은 대조군에 비해 4일에 유의하게 높았다. 기관지폐포세척액에서 산화질소산물은 4일에 가장 높았다. 면역화학염색에서 AQP1과 AQP4는 4일에 감소하였으며 AQP4는 7일에 증가하였고, AQP5는 4, 7, 14일에 발현이 증가하였다. 결 론: 아쿠아포린은 급성폐손상 모델에서 다양하게 발현하여 급성 폐손상의 경과에 따라 다르게 작용함을 시사한다.

Keywords

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;152:1818-24 https://doi.org/10.1164/ajrccm.152.6.8520742
  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 https://doi.org/10.1016/1043-4666(93)90024-Y
  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; 57:782-7 https://doi.org/10.1002/jlb.57.5.782
  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;170:655-63 https://doi.org/10.1084/jem.170.3.655
  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;97:2183-91 https://doi.org/10.1172/JCI118659
  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:C1549-61 https://doi.org/10.1152/ajpcell.1997.273.5.C1549
  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 https://doi.org/10.1165/ajrcmb.22.1.3818
  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 https://doi.org/10.1016/0003-2697(85)90442-7
  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;6:647-56 https://doi.org/10.1016/1043-4666(94)90053-1
  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;33:319-27 https://doi.org/10.1165/rcmb.F305
  12. Ashbaugh DG, Bigelow DB, Petty TL, Levine BE. Acute respiratory distress in adults. Lancet 1967;2:319–23
  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;342:1301–8
  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;351:327-36 https://doi.org/10.1056/NEJMoa032193
  15. Zimmerman GA, Albertine KH, McIntyre TM. Pathogenesis of sepsis and septic-induced lung inury. In: Lenfant C, Matthay MA, editors. Acute respiratory distress syndrome. 1st ed. New York: Marcel Dekker; 2003. p. 245–87
  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;30:489-95 https://doi.org/10.1007/s00134-003-2129-9
  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;38:1709-15 https://doi.org/10.1016/0006-2952(89)90403-6
  18. Moncada S, Palmer RMJ, Higgs EA. Nitric oxide: physiology, pathophysiology and pharmacology. Pharmacol Rev 1991;43:109-42
  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;94:2407-13 https://doi.org/10.1172/JCI117607
  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;262:17497-503
  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;263:15634-42
  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;88:11110-4 https://doi.org/10.1073/pnas.88.24.11110
  23. Preston GM, Carroll TP, Guggino WB, Agre P. Appearance of water channels in Xenopus oocytes expressing red cell CHIP28 protein. Science 1992;256:385-7. https://doi.org/10.1126/science.256.5055.385
  24. Jung JS, Preston GM, Smith BL, Guggino WB, Agre P. Molecular structure of the water channel through aquaporin CHIP. J Biol Chem 1994;269:14648-54
  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;165:243-54 https://doi.org/10.1007/s002329900438
  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;271:4577-80 https://doi.org/10.1074/jbc.271.9.4577