KISEP Rhinology Korean J Otolaryngol 000;:9500 오존노출에의한백서비점막의형태학적변화 정진욱 김용민 나기상 김병국 박찬일 Morphologic Changes in Nasal Mucosa of the Rat after Ozone Exposure Jin Uk Jeong, MD, Yong Min Kim, MD, Ki Sang Rha, MD, Byung Kook Kim, MD and Chan Il Park, MD Department of Otolaryngology, College of Medicine, Chungnam National University, Taejon, Korea ABSTRACT Background and ObjectivesOzone is the principal oxidant pollutant in photochemical smog. Although ozoneinduced pulmonary changes have been morphologically characterized, effects of ozone on the upper respiratory tract have not been thoroughly investigated. The purpose of this study was to investigate the morphological change of nasal mucosa after acute exposure to ozone. Material and MethodsRats are exposed to 0., 0.6, 0.9, and. ppm of O for 6 hours a day for 7 days. Rats were sacrificed 8h after the end of their last exposure. Nasal mucosa was processed for HematoxylinEosin HE stain and Alcian blue ph.5periodic Acid Schiff ABPAS stain. ResultsMorphological changes such as infiltration of neutrophils, loss of cilia, epithelial hyperplasia, squamous metaplasia and secretory cell metaplasia were observed and these changes were more pronounced in the group exposed to high concentration of ozone, compared with the group exposed to low concentration. ConclusionThe results of this study may suggest that ozone exposure induces inflammatory, proliferative, and metaplastic responses in the nasal mucosa. Korean J Otolaryngol 000;:9500 KEY WORDSOzone Nasal mucosa Morphologic changes. 9
9 Fig.. Equipment for ozone exposures. Ozone generator, Air pump Exposure chamber, Charcoal filter, Ozone analyzer. Korean J Otolaryngol 000;:9500
정진욱 외 마리에서 중등도 이상의 비후를 관찰할 수 있었다(Table 섬모의 손상, Fig. ). 0.9 ppm 폭로군과. ppm 폭로군에서는 각각 0. ppm 폭로군에서는 비중격점막에서의 섬모의 손상을 마리의 maxilloturbinate에서 중등도의 비후소견이 관찰 관찰할 수 없었으나, 0.6 ppm 폭로군에서는 개체에서 경 되었다(Table, Fig. ). 도의 손상이 관찰되었으며,. ppm 폭로군에서는 모든 개 체에서 섬모의 손상이 관찰되었다(Table, Fig. ). A B C D Fig.. Light micrographs of normal septal mucosa (A, C) and maxilloturbinate mucosa (B, D). H & E stain (A, B), AB/PAS stain (C, D). (original magnification 00). Table. Histologic changes in the maxilloturbinate or septal mucosa 0. ppm M 0.6 ppm S M 0.9 ppm. ppm S M S M S Neutrophil infiltration Epithelial hyperplasia Increase of secretory cells 5 MT maxilloturbinate, S nasal septum 95
오존노출에 의한 백서 비점막의 형태학적 변화 A B a b c d a b c d Fig.. Light micrographs of septal (A) and maxilloturbinate mucosa (B) of 0. (a), 0.6 (b), 0.9 (c), and. ppm (d) exposure groups. Infiltration of inflammatory cells, ciliary loss, and epithelial hyperplasia were more pronounced in high concentrationexposure groups (original magnification 00). 96 Korean J Otolaryngol 000;:9500
정진욱 외 은 증가된 소견을 보였다. 정상 maxilloturbinate에서는 보 편평상피화생 0. ppm 폭로군에서는 마리에서 경도, 0.6 ppm 폭로군 에서는 마리에서 중등도의 편평상피화생이 관찰되었으나, 이지 않았던 분비세포가 각 군의 5마리중 ~마리에서 관 찰되었다(Table, Fig. 5). 0.9 ppm 폭로군에서는 마리,. ppm 폭로군에서는 모든 개 고 체에서 경도 이상의 편평상피화생을 보였다(Table, Fig. ). 분비세포의 비후 및 증가 찰 외부로부터 해로운 자극 물질에 대하여 방어 작용을 하 0. ppm 폭로군에서 비중격의 분비세포는 대조군과 차 는 것으로 알려진 비점막은 대기오염 물질에 노출되었을 때 이가 없었으나, 0.6 ppm 폭로군 중 마리, 0.9 ppm 폭로 점막내 염증세포, 특히 중성구가 증가하는 것으로 알려져 군 및. ppm 폭로군 중 마리에서 분비세포가 비후 혹 있다.) Hotchkiss 등)은 백서를 각각 0., 0.8,.5 ppm 의 오존농도에 폭로시켰을 때 모든 군에서 비세척액 및 비 Table. Histologic changes in the septal mucosa 점막에서 중성구가 증가되어 있음을 보고하였다. 0. ppm 0.6 ppm 0.9 ppm. ppm 5 Hotchkiss 등5)은 0.5 ppm의 오존에 백서를 8시간 동안 폭로시켰을 때 점막의 다른 변화 보다도 중성구의 침윤이 Loss of cilia 본 연구에서도 오존에 폭로된 모든 군에서 중성구의 침 윤이 증가되어 있었고 농도가 증가할수록 중성구의 침윤정 Squamous metaplasia 도는 심해지는 경향을 관찰할 수 있었다. 가장 먼저 관찰된다고 하였다. 이러한 중성구의 증가가 방 어작용의 일환으로 생각되지만 침윤된 중성구로부터 생산되 는 단백분해효소나 세포에 독성을 가진 산화 유리기로 인 해 상피세포가 손상되어 상피의 괴사 혹은 탈락이 일어날 수 있다.6) A B C D Fig.. Light micrographs of septal mucosa showing mild degree of squamous metaplasia in 0.6 ppm (b) group, moderate degree in 0.9 ppm (c) group, severe degree in. ppm (d) exposure groups compared with 0. ppm (a) group (original magnification 00). 97
오존노출에 의한 백서 비점막의 형태학적 변화 A B a b c d a b c d Fig. 5. Light micrographs of septal mucosa(a) and maxilloturbinate mucosa (B) of 0. (a), 0.6 (b), 0.9 (c), and. ppm (d) exposure group (AB/PAS stain, original magnification 00). 98 Korean J Otolaryngol 000;:9500
99
REFERENCES ) Wardlaw AJ. The role of air pollution in asthma. Clin Exp Allergy 99896. ) Wardlaw AJ. Human laboratory studies of the health effects of air pollutants. Clin Exp Allergy 9955suppl 6. ) Beckett WS. Ozone, air pollution, and respiratory health. Yale J Biol Med 9966775. ) Henderson RF, Hotchkiss JA, Chang IY, Harkema JR. Effect of cumulative exposure to ozone. Toxicol Appl Pharmacol 999 5965. 5) U.S. EPA. National air quality and emission report. Report no. EPA 5R90. Washington U.S. Environmental Protection Agency99. 6) Schwartz LW, Dungworth DL, Mustafa MG, Tarkington BK, Tyler WS. Pulmonary responses of rats to ambient levels of ozone. Effects of 7day intermittent or continuous exposure. Lab Invest 97656578. 7) Plopper CG, Chu FP, Haselton CJ, Peake J, Wu R, Pinkerton KE. Dosedependent tolerance to ozone. I. Tracheobronchial epithelial reorganization in rats after 0 months exposure. Am J Pathol 990. 8) Silverman F. Asthma and respiratory irritants ozone. Environ Health Perspect 97996. 9) Folinsbee LJ, Bedi JF, Horvarth SM. Pulmonary function changes after hour continuous heavy exercise. Am Rev Respir Dis 98 57:988. 0) Hackney JD, Linn WS. Environmental factors and asthma Air pollution and weather. In Weiss EB, Segal MS, Stein M, eds. Bronchial asthma. Mechanism and therapeutics. nd ed. Boston Little Brown985. p.55. ) Yokoyama E, Frank R. Respiratory uptake of ozone in dog. Arch Environ Health 9758. ) CalderonGarciduenas L, OsornoVelazquez A, BravoAlvarez H, DelgadoChavez R, BarriosMarquez R. Histopathologic changes of the nasal mucosa in southwest Metropolitan Mexico City inhabitants. Am J Pathol 995. ) Leikauf GD, Simpson LG, Santrock J, Zhao Q, AbbinanteNissen J, Zhou S, et al. Airway epithelial cell responses to ozone injury. Environ Health Perspect 9950suppl 95. ) Hotchkiss JA, Harkema JR, Sun JD, Henderson RF. Comparison of acute ozoneinduced nasal and pulmonary inflammatory responses in rats. Toxicol Appl Pharmacol 98998890. 5) Hotchkiss JA, Harkema JR, Johnson NF. Kinetics of nasal epithelial cell and proliferation in F rats following a single exposure to 0.5ppm ozone. Toxicol Appl Pharmacol 997758. 6) Weiss SJ. Tissue destruction by neutrophils. N Engl J Med 989 06576. 7) Harkema JR, Plopper CG, Hyde DM, St. George JA, Wilson DW, Dungworth DL. Response of the macaque nasal epithelium to ambient levels of ozone. A morphologic and morphometric study of the transitional and respiratory epithelium. Am J Pathol 98789. 8) Lemos M, Lichtenfels JFC, Amaro E, Martins MA, King M, Bohm GM, et al. Quantitative pathology of nasal passages in rats exposed to urban levels of air pollution. Environ Res 99668795. 9) Harkema JR, Hotchkiss JA, Henderson RF. Effect of 0. and 0.80ppm ozone on nasal and nasopharyngeal epithelial mucosbstances Quantitative histochemistry. Toxicol Pathol 9897 555. 0) Harkema JR, Barr EB, Hotchkiss JA. Responses of rat nasal epithelium to short and longterm exposures of ozone Image analysis of epithelial injury, adaptation and repair. Microsc Res Tech 99767686. 500 Korean J Otolaryngol 000;:9500