Applied Microscopy

Korean Society of Microscopy (한국현미경학회)
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Neutrophilic Respiratory Burst Contributes to Acute Lung Leak in Rats Given N-nitroso-N-methylurethane

N-nitroso-N-methylurethane으로 유도된 급성 폐손상에서 호중구에 의한 산화성 스트레스의 역할

  • Kim, Seong-Eun (Department of Biology, Youngnam University) ;
  • Kim, Dug-Young (Department of Physiology, Catholic University of Daegu) ;
  • Na, Bo-Kyung (Department of Physiology, Catholic University of Daegu) ;
  • Lee, Young-Man (Department of Physiology, Catholic University of Daegu)
  • 김성은 (영남대학교 이과대학 생물학과) ;
  • 김덕영 (대구가톨릭대학교 의과대학 생리학교실) ;
  • 나보경 (대구가톨릭대학교 의과대학 생리학교실) ;
  • 이영만 (대구가톨릭대학교 의과대학 생리학교실)
  • Published : 2003.03.01
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Abstract

As is well known that N-nitroso-N-methylurethane (NNNMU) causes acute lung injury (ALI) in experimental animals. And ALI caused by NNNMU is very similar to ARDS in human being in its pathology and progress. In its context, we investigated the pathogenetic mechanism of ARDS associated with oxidative stress by neutrophils in Sprague-Dawley rat model of NNNMU-induced ALI. NNNMU had increased lung weight/body weight ratio (L/B ratio), lung myeloperoxidase (MPO) activity, protein content and number of neutrophils in bronchoalveolar fluid (BALF) compared with those of control rat (p<0.001, respectively). In contrast, the amount of pulmonary surfactant in BALF was decreased by NNNMU (p<0.001). Morphologically, light microscopic examination denoted pathological findings such as formation of hyaline membrane, infiltration of neutrophils and perivascular cuffing in the lungs of NNNMU-treated rats. In addition, ultrastructural changes such as the necrosis of endothelial cells, swelling and vacuolization of lamellar bodies of alveolar type II cells, and the degeneration of pulmonary surfactant were identified after treatment of NNNMU. Very interestingly, cerium chloride electron microscopic cytochemistry showed that NNNMU had increased the production of cerrous-peroxide granules in the lung, which signified the increased production of hydrogen peroxide in the lung. Collectively, we conclude that NNNMU causes acute lung leak by the mechanism of neutrophilic oxidative stress of the lung.

급성호흡곤란증후군 (acute respiratory distress syndrome)을 포함한 급성폐손상의 기전을 산화성스트레스와 연관하여 알아보기 위하여 본 연구를 시행하였다. N-nitroso-N-methylurethane (NNNMU)은 실험동물에 있어서 사람에서 보이는 ARDS와 유사한 병리학적인 소견을 보이므로 ARDS의 모델로 사용된다. 본 연구에서는 흰쥐에서 NNNMU로 급성폐손상을 유도한 뒤 이를 Lung weight/Body weight ratio, 폐포세척액(BAL)내의 단백함량을 측정하여 확인하고, 동시에 호중구의 침윤에 의한 산소기형성, 이에 따른 산화성스트레스를 확인하기 위하여 BAL 내의 호중구수의 산정 및 폐장의 MPO 활성도를 측정하였다. 동시에 광학현미경, 전자현미경 및 세포화학적 전자현미경법을 이용하여 호중구에 의한 급성폐손상, 호중구의 침윤 및 미세구조의 변화 및 산소기의 생성을 확인하였다. 대조군에 비하여 NNNMU를 투여한 흰쥐에서는 BAL 내의 단백질이 증가하고 BAL 내의 호중구의 증가, 폐장의 MPO 활성도의 증가로 호중구의 침윤이 증가함을 관찰하였다. 광학현미경상 호중구의 침윤, 폐포내 호중구의 유입 및 vascular cuffing 등이 관찰되었다. 전자현미경 소견상 제2형 폐포세포는 산화성 스트레스의 전형적 소견을 보이고 조직의 손상은 호중구에 의한 손상으로 생각되었으며 세포화학적 전자현미경법으로 산소기의 생성이 증가됨을 확인하였다. 이러한 결과들을 종합할 때 NNNMU에 의한 급성폐손상은 호중구의 산소기 생성에 의한 산화성스트레스에 의한 것으로 생각되었다.

Keywords

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