Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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The Change of Cell Distribution in the lung and the Expression Pattern of IL-4 and IL-10 in Asthma Induced Mouse

천식유발 마우스에서의 폐 내 세포조성 변화와 IL-4 및 IL-10의 발현 양상

  • Lee, Soo-Jin (Konkuk University College of Veterinary Medicine) ;
  • Park, Se-Jong (Konkuk University College of Veterinary Medicine) ;
  • Li, Tian-Zhu (Konkuk University College of Veterinary Medicine) ;
  • Jang, Yang-Ho (Konkuk University College of Veterinary Medicine) ;
  • Choe, Nong-Hoon (Konkuk University College of Veterinary Medicine)
  • Published : 2006.08.30
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Abstract

Asthma is a chronic inflammatory disorder of the airways, which characterized by bronchial hyperresponsiveness, reversible airflow limitation and respiratory symptoms. Internationally, the prevalence of asthma has been increased over last 3 decades. Recently, several studies of asthma have been reported with gradually increasing importance. To tesify the hypothesis that interleukin (IL)-4 and IL-10 may be an important determinant of the severity of airway inflammation, their expression was studied in mouse model of asthma. BALB/c mouse, IL-4 Knockout (KO) mouse and IL-10 KO mouse were sensitized with intraperitoneal injection of ovalbumin adsorbed to aluminum potassium sulfate, followed by challenges with intranasal ovalbumin on 3 consecutive days. The severity of pulmonary inflammation was assessed by eosinophilia in BAL fluid, number of total BAL cells, histopathological changes in lung tissues, and immunohistochemical staining against IL-4 and IL-10. In BAL fluid, the number of total cells was significantly increased in asthma induced mouse compare to the control. In asthma induced mouse, eosinophil was increased to 56% and neutrophil was 0.2%. In H &E stains, eosinophilic infiltration and epithelium hyperplasia were clearly noticed in asthma induced mouse. In immunohistochemical staining for IL-4 and IL-10, there was no positive reaction in control group. However, very strong reactions were appeared in asthma induced group. In this research, IL-4 and IL-10, which seem to play a central role in allergic asthma, KO mouse was utilized to test the causative relationship between airway inflammation and role of specific cytokine. Asthma induced IL-4 and IL-10 KO mice showed much decreased inflammatory reactions in the number of total BAL cells, in eosinophilic infiltration, and in immunohistochemical stains against diverse inflammatory proteins. These results suggest that IL-4 and IL-10 increase the asthmatic reactions in vivo mice model.

본 연구진은 일차로 BALB/C 마우스를 이용하여 항원으로 ovalbumin을 사용하여 천식을 유발하여 폐 내 세포구성에 미치는 영향을 다른 실험방법에 의해 만들어진 천식모델과 비교하여 살펴보았고 이차로 천식 발생에 중요한 역할을 하는 것으로 알려진 IL-4와 IL-10 knock out (KO) 마우스를 이용하여 천식모델을 구축하여 천식인자를 가진 개체에서의 반응과 천식관련인자가 결핍된 개체에서의 반응성의 차이를 살펴보았다. 천식의 유발은 실험 1일째 $20\;{\mu}g$ ovalbumin으로 감작시킨 후 실험 14일에 재감작을 시켰다. 그 후 nebulizer를 이용하여 nasal inhalation을 28일, 29일, 30일에 실시하여 천식을 유발시켰다. 천식 유발의 확인은 기관지폐포 세척술로 채취한 폐 내 세포액을 이용하여 총 세포수 및 염증세포의 증가와 폐 내 세포와 폐조직의 염색을 통해 분포율을 확인하였다. 천식의 발병 과정에서 IgE 관련 과민반응을 주도하는 IL-4와 Th2 세포의 기능을 억제한다고 알려진 IL-10의 면역화학염색을 통해 그 발현정도를 관찰하였다. BALB/c 마우스의 천식유발군의 경우 천식의 특징인 염증세포의 증가와 호산구의 증가와 세기관지 주위 염증세포의 침윤 및 기도 상피의 비후를 관찰할 수 있었다. 따라서 본 실험에 응용된 천식유발 방법은 적합하였으며 천식유발 절차가 성공적으로 수행되었음을 확인하였다. IL-4 및 IL-10 KO 마우스를 이용한 천식유발군의 경우 BALB/c 마우스를 이용한 천식유발군보다 호산구의 증가 정도가 미약하게 관찰되었고, 폐조직 내 염증세포의 침윤 정도도 감소하였다. 천식매개인자인 IL-4와 IL-10의 면역조직화학염색 결과에서도 양성반응이 거의 나타나지 않았다. 본 연구 결과, IL-10이 IL-4와 같이 천식과 관련하여 세기관지 염증 반응을 증가시키는 역할을 하는 것으로 생각된다.

Keywords

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