Tuberculosis and Respiratory Diseases

The Korean Academy of Tuberculosis and Respiratory Diseases (대한결핵및호흡기학회)
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Cytosolic Phospholipase A2 Activity in Neutrophilic Oxidative Stress of Platelet-activating Factor-induced Acute Lung Injury

Platelet-activating Factor에 의한 급성폐손상에서 호중구성 산화성 스트레스에 관여하는 Cytosolic Phospholipase A2 활성도의 변화

  • Kwon, Young Shik (Department of Physiology, School of Medicine, Daegu Catholic University) ;
  • Hyun, Dae Sung (Department of Internal Medicine, School of Medicine, Daegu Catholic University) ;
  • Lee, Young Man (Department of Physiology, School of Medicine, Daegu Catholic University)
  • 권영식 (대구가톨릭대학교 의과대학 생리학교실) ;
  • 현대성 (대구가톨릭대학교 의과대학 내과학교실) ;
  • 이영만 (대구가톨릭대학교 의과대학 생리학교실)
  • Received : 2007.07.09
  • Accepted : 2007.11.15
  • Published : 2007.12.30
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Abstract

Background: The present investigation was performed in rats and isolated human neutrophils in order to confirm the presumptive role of the positive feedback loop of cytosolic phospholipase $A_2$ ($cPLA_2$) activation by plateletactivating factor (PAF). Methods: The possible formation of the positive feedback loop of the $cPLA_2$ activation and neutrophilic respiratory burst was investigated in vivo and in vitro by measurement of the parameters denoting acute lung injury. In addition, morphological examinations and electron microscopic cytochemistry were performed for the detection of free radicals in the lung. Results: Five hours after intratracheal instillation of PAF ($5{\mu}g/rat$), the lung leak index, lung myeloperoxidase (MPO) activity, the number of neutrophils and the concentration of cytokine-induced neutrophil chemoattractant (CINC) in bronchoalveolar lavage fluid were increased by PAF as compared with those of control rats. The NBT assay and cytochrome-c reduction assay revealed an increased neutrophilic respiratory burst in isolated human neutrophils following exposure to PAF. Lung and neutrophilic $cPLA_2$ activity were increased following PAF exposure and exposure to hydrogen peroxide increased $cPLA_2$ activity in the lung. Histologically, inflammatory findings of the lung were observed after PAF treatment. Remarkably, as determined by $CeCl_3$ cytochemical electron microscopy, increased production of hydrogen peroxide was identified in the lung after PAF treatment. Conclusion: PAF mediates acute oxidative lung injury by the activation of $cPLA_2$, which may provoke the generation of free radicals in neutrophils.

연구배경: 급성호흡곤란증후군의 병인론에 관여하는 PAF의 역할이 다양하고 중요하므로 본 연구에서는 PAF의 또 다른 작용의 가능성, 즉 $cPLA_2$의 활성화(retrograde activation of $cPLA_2$ by PAF)의 가능성을 검사하고자 하였다. 즉, $cPLA_2$의 활성화에 따른 염증성 지질분자의 생성이 산소기의 생성과정을 증폭시키고 이 때 생성된 PAF가 역으로 $cPLA_2$를 활성화시키는지를 확인하기 위하여 본 연구는 고안되었다. 방 법: 흰쥐에서 급성폐손상을 유도하기 위하여 $5{\mu}g$의 PAF를 0.5 ml의 0.25% bovine serum albumin 용액과 혼합한 뒤 기도 내로 직접 분무하거나 0.5 ml의 4.5 mM의 과산화수소를 기도 내로 분무하였다. 대조군의 경우는 0.5 ml의 생리적 식염수를 기도 내로 분무하였다. 5 시간 후에 단백누출지수 측정, 폐장의 MPO 활성도 측정, 폐포 세척액 내의 호중구 산정, CINC 측정, NBT 및 cytochrome-c 환원검사를 시행하였다. 또한 폐장 및 호중구에 서의 $cPLA_2$ 활성도의 측정 및 광학현미경과 전자현미경을 이용하여 형태학적 관찰을 시행하였다. 결 과: PAF투여 후 단백누출지수, MPO, BAL내의 호중 구의 수 및 CINC의 농도가 대조군에 비하여 유의하게 증가하였다. NBT및 cytochrome-c환원검사의 결과 PAF는 호중구의 respiratory burst를 현저히 증가시키고, 분리된 사람의 호중구에서도 산소기의 생성을 현저히 증가시켰 다. 동시에 PAF는 분리된 호중구 및 폐장의 $cPLA_2$의 활성도도 증가 시켰다. 폐장 내로 투여한 과산화수소는 폐장의 $cPLA_2$활성도를 대조군에 비하여 현저히 증가시켰다. 결 론: $cPLA_2$의 활성화에 따라 생성된 PAF는 호중구의 산소기 생성을 증가시켜 폐장 내의 산화성스트레스를 유발하고 동시에 이때 생성된 산화기는 $cPLA_2$를 활성화시키며 PAF 또한 $cPLA_2$의 활성도를 증가시켜 PAF가 급성호흡 곤란증후군의 병인론에 관여하는 것으로 생각된다.

Keywords

References

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