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The Effects of Diesel Exhaust Particles on the Alveolar Macrophages for Inducible Nitric Oxide Synthase Induction and Nitric Oxide with Nitrotyrosilated-protein Formation

디젤분진이 폐포대식세포에서 nitric oxide의 생성과 inducible nitric oxide synthase의 발현 및 nitrotyrosilated-protein의 형성에 미치는 효과

  • Lim Young (Catholic University College of Medicine) ;
  • Choe Myung-Ok (Catholic University College of Medicine) ;
  • Lee Kweon-Haeng (Catholic University College of Medicine) ;
  • Kim Kyung-A (Catholic University College of Medicine) ;
  • Kim Kil-Soo (Kyungpook national University College of Veterinary Medicine) ;
  • Lee Myoung-Heon (National Veterinary Research and Quarantine Service) ;
  • Li Tian-Zhu (Konkuk University College of Veterinary Medicine) ;
  • Lee Soo-Jin (Konkuk University College of Veterinary Medicine) ;
  • Choe Nong-Hoon (Konkuk University College of Veterinary Medicine)
  • Published : 2006.04.01

Abstract

Epidemiological studies have demonstrated an association between exposure to diesel exhaust particles (DEP) and adverse cardiopulmonary effects. Despite the epidemiological proof, the pathogenesis of DEP-related pulmonary diseases remain poorly understood. So, comprehensive in vivo and in vitro researches are required to know the effects of DEP on diverse lung diseases. Alveolar macrophages (AM) and airway epithelial cells are known as important cellular targets in DEP-induced lung diseases. Other studies have shown that nitric oxide (NO) is involved in particle matter induced lung injury. The present study was undertaken to determine whether DEP has an synergistic effects on lipopolysaccharide (LPS)-induced NO formation and inducible nitric oxide synthase (iNOS) with nitrotyrosilated-protein formation in cultured primary alveolar macrophages. The formation of NO was determined through the Griess reaction in the cultured medium and iNOS with nitrotyrosilated-proteins are analyzed by immunohistochemical staining and Western analysis. The results indicate that DEP exposure does not induce NO formation by itself, however DEP showed significant synergistic effects on LPS-induced NO formation. So, our results suggest that DEP inhalation could aggravate inflammatory lung disease through NO formation.

본 연구에서는 DEP의 노출이 새로운 호흡기계 질환 유발의 가능성과 호흡기계의 염증성인자로 잘 알려진 lipopolysaccharide (LPS)의 역할에 어떠한 영향을 미치는 지를 확인하고자 폐에서 염증성 반응 시 생성이 증가하는 것으로 알려진 Nitric Oxide (NO)의 형성과 NO의 생성에 관련된 효소인 inducible nitric oxide synthase (iNOS) 및 NO에 의하여 형성되는 것으로 알려진 nitrotyrosilated-protein을 폐포 대식세포를 통해 분석하였다. 폐포대식세포에 DEP를 농도 별로 단독 처리하였을 때와 동일한 농도에서 배양시간을 달리하였을 때는 NO가 생성되지 않았으나 DEP와 함께 LPS를 처리하였을 때는 LPS를 단독으로 처리했을 때보다. 유의성이 있게 증가함을 확인할 수 있었다. 또한 NO의 생성에 관련된 효소인 iNOS 및 NO에 의하여 형성되는 것으로 알려진 nitrotyrosilated-protein 발현의 정도를 면역화학염색과 Western analysis로 확인할 수 있었다. DEP는 폐포대식세포에서 직접적으로 NO생성에 영향을 미치지 않았으며, NO를 생성하는 iNOS나 nitrotyrosilated-protein의 발현에도 영향을 주지 않았으나 세균성 염증인자의 한 종류인 LPS가 NO를 형성하는 데에는 통계학적인 상승효과가 있었다. 결론적으로 본 연구에서는 염증성질환의 환자에서 DEP의 흡입은 간접적으로 NO를 형성하는데 영향을 미쳐 질환을 악화시킬 것으로 판단한다.

Keywords

References

  1. Al-Humadi N. H., P. D. Siegel, D. M. Lewis, M. W. Barger, J. Y. Ma, D. N. Weissman and J. K. Ma. 2002. Alteration of intracellular cysteine and glutathione levels in alveolar macrophages and lymphocytes by diesel exhaust particle exposure. Environ. Health Perspect 110, 349-353 https://doi.org/10.1289/ehp.02110349
  2. Bascom R., P. A. Bromberg and D. L. Costa. 1996. Health effects of outdoor air pollution. Am. J. Respir. Crit. Care Med. 153, 477-498 https://doi.org/10.1164/ajrccm.153.2.8564086
  3. Bunn W. B. III, P. A. Valberg, T. J. Slavin and C. A. Lapin. 2002. What is new in diesel. Int. Arch. Occup. Environ. Health 75(Suppl.), S122-S132 https://doi.org/10.1007/s00420-002-0342-4
  4. Castranova V., J. Y. Ma, H. M. Yang, J. M. Antonini, L. Butterworth, M. W. Barger, J. Roberts and J. K. Ma. 2001. Effect of Exposure to Diesel Exhaust Particles on Susceptibility of the lung to infection. Environ. Health Perspect I109(suppl.), 609-612
  5. Cheng, Y. S., H. C. Yeh, J. L. Mauderley and B. V. Mokler. 1984. Characterization of diesel exhaust in a chronic inhalation study. Am. Ind. Hyg. Assoc. J. 45, 547-555 https://doi.org/10.1080/15298668491400241
  6. Choe, N. H., S. Tanaka and E. Kagan. 1998. Asbestos fiber and interleukin-1 upregulate the formation of reactive nitrogen species in rat pleural mesothelial cells. Am. Respir. Cell. Mol. Bol. 19, 226-236 https://doi.org/10.1165/ajrcmb.19.2.3111
  7. Churg, A. and M. Brauer. 1997. Human lung parenchyma retains PM 2.5. Am. J. Respir. Cri. Care Med. 155, 2109-2111 https://doi.org/10.1164/ajrccm.155.6.9196123
  8. Coleman, J. W. 2001. Nitric oxide in immunity and inflammation. Int. Immunopharmacol. 1, 1397-1406 https://doi.org/10.1016/S1567-5769(01)00086-8
  9. Committee of the Environmental and Occupational Health Assembly of the American Thoracic Society. 1996. Health effects of outdoor air pollution. Am. J. Respir. Crit. Care Med. 153, 3-50 https://doi.org/10.1164/ajrccm.153.1.8542133
  10. Dockery, D. W. and C. A. III. Pope. 1994. Acute respiratory effects of particulate air pollution. Annu. Rev. Public Health 15, 107-132 https://doi.org/10.1146/annurev.pu.15.050194.000543
  11. Emanuel, M. B. 1998. Hay fever, a post industrial revolution epidemic: a history of its growth during the 19th century. Clin. Allergy 18, 295-304
  12. Environmental Protection Agency, Office of Air Quality Planning and Standards. 1996. Review of the National Ambient Air Quality Standards for Particulate Matter: policy assessment of scientific and technical information. OAQPS staff paper no. EPA-452\R-96-013
  13. Fels, A. O. and Z. A. Cohn. 1986. The alveolar macrophages. J. Appl. Physiol. 60, 355-369
  14. Handa, T., T. Yamauchi, M. Ohnishi, Y. Hisamatu and T. Ishii. 1983. Detection and average content levels of carcinogenic and mutagenic compounds from the particulate on diesel and gasoline engine mufflers. Environ. Int. 9, 335-341 https://doi.org/10.1016/0160-4120(83)90123-X
  15. Harre, E. S. M., P. D. Price and R. B. Ayrey. 1997. Respiratory effects of air pollution in chronic obstructive pulmonary disease: a three-month prospective study. Thorax 52, 1040-44 https://doi.org/10.1136/thx.52.12.1040
  16. Ito, T., M. Ikeda, H. Yamsaki, M. Sagai and T. Tomita. 2000. Peroxinitrite formation by diesel exhaust particles in alveolar cells: Links to pulmonary inflammation. Environ. Toxicol. and Pharmacol. 9, 1-8 https://doi.org/10.1016/S1382-6689(00)00053-3
  17. Kagan, E., Y. Oghiso and D. P. Hartmann. 1983. The effects of chrysotile and crocidolite asbestos on the lower respiratory tract: analysis of bronchoalveolar lavage constituents. Environ. Res. 32, 382-397 https://doi.org/10.1016/0013-9351(83)90120-2
  18. Lebowitz, M. D. 1996. Epidemiological studies of the respiratory effects of air pollution. Eur. Respir. J. 9, 1029-1054 https://doi.org/10.1183/09031936.96.09051029
  19. Peterson, B. and A. Saxon. 1996. Global increases in allergic respiratory disease: the possible role of diesel exhaust particles. Ann. Allergy Asthma Immunol. 77, 263-270 https://doi.org/10.1016/S1081-1206(10)63318-2
  20. Pope, C. A. III, D. W. Dockery, J. D. Spengler and M. E. Raizenne. 1991. Respiratory health and PM10 pollution. A daily time series analysis. Am. Rev. Respir. Dis. 144, 668-674 https://doi.org/10.1164/ajrccm/144.3_Pt_1.668
  21. Pope, C. A. III and R. E. Kanner. 1993. Acute effects of PM10 pollution on pulmonary function of smokers with mild to moderate COPD. Am. Rev. Respir. Dis. 147, 1336-1340 https://doi.org/10.1164/ajrccm/147.6_Pt_1.1336
  22. Pope, C. A. III, M. J. Thun, M. M. Namboodiri, D. W. Dockery, J. S. Evans, F. E. Speizer and C. W. Jr. Heath. 1995. Particulate air pollution as predictor of mortality in a prospective study of U. S. adults. Am. J. Respir. Crit. Care. Med. 151, 669-674 https://doi.org/10.1164/ajrccm/151.3_Pt_1.669
  23. Schuetzle, D. and J. A. Frazier. 1986. Factors influencing the emission of vapor and particulate phase components from diesel engines. Dev. Toxicol. Environ. Sci. 13, 41-63
  24. Tanaka, S., N. Choe, D. R. Hemenway, S. Zhu, S. Matalon and E. Kagan. 1998. Asbestos inhalation induces reactive nitrogen species and nitrotyrosine formation in the lungs and pleura of the rat. J. Clin. Invest. 102, 445-54 https://doi.org/10.1172/JCI3169
  25. Takizawa, H., T. Ohtoshi, S. Kawasaki, S. Abe, I. Sugawara, K. Nakahara, K. Matsushima and S. Kudoh. 2000. Diesel exhaust particles activate human bronchial epithelial cells to express inflammatory mediators in the airways: A review. Respirology 5(2), 197-203 https://doi.org/10.1046/j.1440-1843.2000.00245.x
  26. Thomas, G. and P. W. Ramwell. 1988. Vasodilatory properties of mono -L-arginine-containing compounds. Biochem. Biophys. Res. Commun. 154, 332-338 https://doi.org/10.1016/0006-291X(88)90689-4
  27. Yin, X. J., R. Schafer, J. Y. Ma, J. M. Antonini, D. D. Weissman, P. D. Siegel, M. W. Barger, J. R. Roberts and J. K. Ma. 2002. Alteration of pulmonary immunity to Listeria monocytogenes by diesel exhaust particles (DEPs). I. Effects of DEPs on early pulmonary responses. Environ. Health Perspect 11, 1105-1111