Tuberculosis and Respiratory Diseases

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

Expression of Matrix Metalloproteinase-9 and Tissue Inhibitor of Metalloproteinase-1 after Administration of Endotoxin in Diabetic Rats

내독소로 자극된 당뇨 쥐에서 단백분해효소와 그 억제제 발현

  • Seo, Ki Hyun (Department of Internal Medicine, Soonchunhyang University College of Medicine, Clinical Research institute) ;
  • Choi, Jae Sung (Department of Internal Medicine, Soonchunhyang University College of Medicine, Clinical Research institute) ;
  • Na, Joo Ok (Department of Internal Medicine, Soonchunhyang University College of Medicine, Clinical Research institute) ;
  • Uh, Soo Taek (Department of Internal Medicine, Soonchunhyang University College of Medicine, Clinical Research institute) ;
  • Kim, Yong Hoon (Department of Internal Medicine, Soonchunhyang University College of Medicine, Clinical Research institute) ;
  • Park, Choon Sik (Department of Internal Medicine, Soonchunhyang University College of Medicine, Clinical Research institute)
  • 서기현 (순천향대학교 의과대학 내과학교실, 임상의학연구소) ;
  • 최재성 (순천향대학교 의과대학 내과학교실, 임상의학연구소) ;
  • 나주옥 (순천향대학교 의과대학 내과학교실, 임상의학연구소) ;
  • 어수택 (순천향대학교 의과대학 내과학교실, 임상의학연구소) ;
  • 김용훈 (순천향대학교 의과대학 내과학교실, 임상의학연구소) ;
  • 박춘식 (순천향대학교 의과대학 내과학교실, 임상의학연구소)
  • Received : 2006.04.24
  • Accepted : 2006.09.04
  • Published : 2006.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

Background: An acute lung injury(ALI) is characterized by the recruitment, activation, and apoptosis of inflammatory cells, numerous products released by inflammatory cells such as reactive oxygen species, inflammatory mediators, and a variety of proteolytic enzymes. It was reported that bacterial infections in diabetics showed impaired PMN functions such as reduced PMN respiratory burst and decreased microbicidal activity in inflamed tissue. However, the effect of the proteinase - inhibitor (MMP-9 vs TIMP-1) in ALI in diabetics is unclear. This study evaluated the differences in the expression of MMP-9 and TIMP-1 after the stimulation of endotoxin in a rat model. Methods: Six-week-old male Sprague-Dawley rats were classified into normal, DM, LPS and DM+LPS groups. The peripheral blood, BAL fluids, and lung tissues were obtained from individual rats. The MMP-9 activity was measured by gelatin zymography and the TIMP-1 level was measured by Western blotting. Results: The total BAL cells of the DM-LPS groups were significantly lower than the LPS groups (p < 0.01). The MMP-9 activities in the serum were higher in the DM+LPS groups than in the other groups. The MMP-9 activities in the BAL fluids were significantly higher in the DM+LPS group than in the normal and diabetic rats (p < 0.05). TIMP-1 expressions in the BAL fluids were significantly lower in the DM+LPS group than other groups (p < 0.05). The ratio between MMP-9 and TIMP-1 in the BAL fluids was significantly higher in the DM+LPS groups (p < 0.05). Conclusion: In ALI in diabetics the higher MMP-9 activity and lower TIMP-1 level are believed to prolonged and intensify the course of inflammation.

연구배경: 급성폐손상에서 염증세포의 모집과 활성, 그 염증세포의 부산물인 활성산소, 사이토카인, 기질단백분해효소등과 기획성 세포사멸이 폐실질 파괴와 재구성에 중요한 역할을 하는 것으로 알려져 있다. 또한 당뇨환자에서 세균 감염 시 중성구의 기능 이상과 활성 산화기의 기능이상, 살균 능의 저하를 가져온다고 알려져 있으나 단백분해효소 대한 견해가 부족한 편이다. 본 연구에서는 동물 모델에서 내독소로 유발된 급성폐손상의 기전에 중요한 역할을 하는 단백분해효소와 그 억제제의 분비를 비교 관찰하였다. 방 법: 생후 6주된 수컷 쥐 40마리를 정상군, 당뇨군, 내독소군, 당뇨-내독소군으로 나누어 당뇨군은 streptozotocin을 투여하였고 내독소군은 지질다당질을 투여하고서 희생시켰다. 각각 혈액, 기관지폐포세척액을 얻어 gelatin zymography를 이용하여 MMP-9의 활성을 측정하였고 Western blot으로 TIMP-1을 측정하였다. 결 과: 1) 내독소군과 당뇨-내독소군은 정상과 당뇨군에 비해 폐 무게의 증가와 기관지폐포세척액과 폐 조직 소견상 염증세포, 특히 중성구의 증가를 보였고 기관지폐포세척액 내 단백질 양의 증가를 보였다 (p < 0.05). 또한 당뇨-내독소군은 내독소군과 달리 기관지폐포세척액 내 총 염증세포 수가 유의하게 감소하였다 (p = 0.001). 2) MMP-9의 활성은 혈청에서는 당뇨-내독소군이 다른 세 군보다 가장 높은 활성을 보였고 기관지폐포세척액에서는 내독소군과 당뇨-내독소군은 차이가 없었으나 정상과 당뇨군에 비해 높은 활성을 보였다 (p < 0.05). TIMP-1 발현은 혈청에서는 네 군간의 차이가 없었고 기관지폐포세척액은 당뇨-내독소군이 가장 낮은 발현을 보였다 (p < 0.05). MMP-9/ TIMP-1 density ratio를 비교했을 때 기관지폐포세척액에서 당뇨-내독소군이 다른 세 군보다 높았고 내독소군도 정상과 당뇨군보다 높았다 (p < 0.05). 결 론: 급성폐손상이 있는 당뇨에서는 염증세포의 화학 주성 감소로 중증 감염에 민감해지고, 기질단백분해효소 활성 증가와 그 억제제의 감소로 폐손상이 가중될 것으로 예상된다.

Keywords

References

  1. Hanley ME, Repine JE. Pathogenesis aspects of the adult respiratory distress syndrome. Semin Respir Crit Care Med 1994:15:260-70 https://doi.org/10.1055/s-2007-1006372
  2. Holter JF, Weiland JE, Patcht ER, Gadek JE, Davis WB. Protein permeability in the adult respiratory distress syndrome: loss of size selectivity of the alveolar epithelium. J Clin Invest 1986:78:1513-22 https://doi.org/10.1172/JCI112743
  3. Simon RH, DeHart PD, Todd RF 3rd. Neutrophilinduced injury of rat pulmonary alveolar epithelial cells. J Clin Invest 1986:78:1375-86 https://doi.org/10.1172/JCI112724
  4. Smedly LA, Tonnesen MG, Sandhaus RA, Haslett C, Guthrie LA, Johnston RB Jr, et al. Neutrophilmediated injury to endothelial cells: enhancement by endotoxin and essential role of neutrophil elastase. J Clin Invest 1986:77:1233-43 https://doi.org/10.1172/JCI112426
  5. Katzenstein AL, Bloor CM, Liebow AA. Diffuse alveoar damage: the role of oxygen, shock and related factors. Am J Pathol 1976:85:209-28
  6. Palmgren MS, deShazo RD, Carter RM, Zimny ML, Shah SV. Mechanisms of neutrophil damage to human alveolar extracelluar matrix: the role of serine and metalloproteinases. J Allergy Clin Immunol 1992:89:905-15 https://doi.org/10.1016/0091-6749(92)90447-A
  7. Shapiro SD, Senior RM. Matrix metalloproteinases: matrix degradation and more. Am J Respir Cell Mol Biol 1999:20:1100-2 https://doi.org/10.1165/ajrcmb.20.6.f151
  8. Matute-Bello G, Liles WC, Radella F 2nd, Steinberg KP, Ruzinski JT, Jonas M, et al. Neutrophil apoptosis in the acute respiratory distress syndrome. Am J Respir Crit Care Med 1997:156:1969-77 https://doi.org/10.1164/ajrccm.156.6.96-12081
  9. van den Steen PE, Proost P, Wuyts A, van Damme J, Opdenakker G. Neutrophil gelatinase B potentiates interleukin-8 tenfold by aminoterminal processing, whereas it degrades CTAP-III, PF-4, and GRO-alpha and leaves RANTES and MCP-2 intact. Blood 2000:96:2673-81
  10. Gibbs DF, Shanley TP, Warner RL, Murphy HS, Varani J, Johnson KJ. Role of matrix metalloproteinases in models of macrophage-dependent acute lung injury: evidence for alveolar macrophage as source of proteinases. Am J Respir Cell Mol Biol 1999:20:1145-54 https://doi.org/10.1165/ajrcmb.20.6.3482
  11. Atkinson JJ, Senior RM. Matrix metalloproteinase-9 in lung remodeling. Am J Respir Cell Mol Biol 2003:28:12-24 https://doi.org/10.1165/rcmb.2002-0166TR
  12. Stetler-Stevenson WG. Dynamics of matrix turnover during pathologic remodeling of the extracellular matrix. Am J Pathol 1996:148:1345-50
  13. Lanchou J, Corbel M, Tanguy M, Germain N, Boichot E, Theret N, et al. Imbalance between matrix metalloproteinases(MMP-9 and MMP-2) and tissue inhibitors of metalloproteinases(TIMP-1 and TIMP-2) in acute respiratory distress syndrome patients. Crit Care Med 2003:31:536-42 https://doi.org/10.1097/01.CCM.0000048626.02184.F8
  14. Joshi N, Caputo GM, Weitekamp MR, Karchmer AW. Infections in patients with diabetes mellitus. N Engl J Med 1999:341:1906-12 https://doi.org/10.1056/NEJM199912163412507
  15. Lippmann ML, Goldberg SK, Walkenstein MD, Herring W, Gordon M. Bacteremic pneumococcal pneumonia: a community hospital experience. Chest 1995:108:1608-13 https://doi.org/10.1378/chest.108.6.1608
  16. Chen KY, Hsueh PR, Liaw YS, Yang PC, Luh KT. A 10-year experience with bacteriology of acute thoracic empyema. Chest 2000:117:1685-9 https://doi.org/10.1378/chest.117.6.1685
  17. Bashar M, Alcabes P, Rom WN, Condos R. Increased incidence of multidrug-resistant tuberculosis in diabetic patients on the Bellevue chest service, 1987 to 1997. Chest 2001:120:1514-9 https://doi.org/10.1378/chest.120.5.1514
  18. Goldman MD. Lung dysfunction in diabetes. Diabetes Care 2003:26:1915-8 https://doi.org/10.2337/diacare.26.6.1915
  19. Becher G, Winsel K, Beck E, Neubauer G, Stresemann E. Breath condensate as a method of noninvasive assessment of inflammation mediators from the lower airways. Pneumologie 1997:51(Suppl 2):456-9
  20. Enomoto T, Usuki J, Azuma A, Nakagawa T, Kudoh S. Diabetes mellitus may increase risk for idiopathic pulmonary fibrosis. Chest 2003:123;2007-11 https://doi.org/10.1378/chest.123.6.2007
  21. Marhoffer W, Stein M, Maeser E, Federlin K. Impairment of polymorphonuclear leukocyte function and metabolic control of diabetes. Diabetes Care 1992:15:256-60 https://doi.org/10.2337/diacare.15.2.256
  22. Shah SV, Wallin JD, Eilen SD. Chemiluminescence and superoxide anion production by leukocytes from diabetic patients. J Clin Endocrinol Metab 1983:57:402-9 https://doi.org/10.1210/jcem-57-2-402
  23. Anderson B, Goldsmith GH, Spagnuolo PJ. Neutrophil adhesive dysfunction in diabetes mellitus: the role of cellular and plasma factors. J Lab Clin Med 1988:111:275-85
  24. Delamaire M, Maugendre D, Moreno M, le Goff MC, Allannic H, Genetet B. Impaired leukocyte functions in diabetic patients. Diabet Med 1997:14:29-34 https://doi.org/10.1002/(SICI)1096-9136(199701)14:1<29::AID-DIA300>3.0.CO;2-V
  25. Dandona P, Aljada A, Mohanty P, Ghanim H, Bandyopadhyay A, Chaudhuri A. Insulin suppresses plasma concentration of vascular endothelial growth factor and matrix metalloproteinase-9. Diabetes Care 2003:26:3310-4 https://doi.org/10.2337/diacare.26.12.3310
  26. Death AK, Fisher EJ, McGrath KC, Yue DK. High glucose alters matrix metalloproteinase expression in two key vascular cells: potential impact on atherosclerosis in diabetes. Atherosclerosis 2003:168: 263-9 https://doi.org/10.1016/S0021-9150(03)00140-0
  27. Lee YC, Lee HB, Rhee YK, Song CH. The involvement of matrix metalloproteinase-9 in airway inflammation of patients with acute asthma. Clin Exp Allergy 2001:31:1623-30 https://doi.org/10.1046/j.1365-2222.2001.01211.x
  28. Hayashi T, Stetler-Stevenson WG, Fleming MV, Fishback N, Koss MN, Liotta LA, et al. Immunohistochemical study of metalloproteinases and their tissue inhibitor in lungs of patients with diffuse alveolar damage and idiopathic pulmonary fibrosis. Am J Pathol 1996:149:1241-56
  29. Ricou B, Nicod L, Lacraz S, Welgus HG, Suter PM, Dayer JM. Matrix metalloproteinases and TIMP in acute respiratory distress syndrome. Am J Respir Crit Care Med 1996:154:346-52 https://doi.org/10.1164/ajrccm.154.2.8756805
  30. Finlay GA, O'Driscoll LR, Russell KJ, D'Arcy EM, Masterson JB, FitzGerald MX, et al. Matrix metalloproteinases expression and production by alveolar macrophages in emphysema. Am J Respir Crit Care Med 1997:156:240-7 https://doi.org/10.1164/ajrccm.156.1.9612018
  31. Gunther A, Walmrath D, Grimminger F, Seeger W. Pathophysiology of acute lung injury. Semin Respir Crit Care Med 2001:22:247-58 https://doi.org/10.1055/s-2001-15782
  32. Pecsvarady Z, Fisher TC, Darwin CH, Fabok A, Maqueda TS, Saad MF, et al. Decreased polymorphonuclear leukocyte deformability in NIDDM. Diabetes Care 1994:17:57-63 https://doi.org/10.2337/diacare.17.1.57