Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Effects of Pseudomonas fluorescens on Production of Several Inflammatory Mediators in the Human Alveolar Epithelial Cells.

재조합 단백질 생산에 이용되는 Pseudomonas fluorescens의 인체 폐포 상피세포의 염증성 인자들의 발현에 미치는 영향

  • Yang, Hyun (Department of Microbiology and Immunology, Medical Research Institute, Pusan National University School of Medicine) ;
  • Ryoo, Jung-Min (Korea science academy) ;
  • Park, Seung-Hwan (Department of Microbiology and Immunology, Medical Research Institute, Pusan National University School of Medicine) ;
  • Choi, Hye-Jin (Department of Microbiology and Immunology, Medical Research Institute, Pusan National University School of Medicine) ;
  • Kim, Na-Yeon (Korea science academy) ;
  • Cho, Hyung-Hoon (Korea science academy) ;
  • Ahn, Jung-Hoon (Korea science academy) ;
  • Moon, Yu-Seok (Department of Microbiology and Immunology, Medical Research Institute, Pusan National University School of Medicine)
  • 양현 (부산대학교 의학전문대학원 미생물학 및 면역학교실) ;
  • 류정민 (한국과학영재학교) ;
  • 박성환 (부산대학교 의학전문대학원 미생물학 및 면역학교실) ;
  • 최혜진 (부산대학교 의학전문대학원 미생물학 및 면역학교실) ;
  • 김나연 (한국과학영재학교) ;
  • 조형훈 (한국과학영재학교) ;
  • 안정훈 (한국과학영재학교) ;
  • 문유석 (부산대학교 의학전문대학원 미생물학 및 면역학교실)
  • Published : 2008.04.30
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Abstract

To investigate the molecular mechanism of the airway inflammation by Pseudomonas fluorescens, effects on the inflammatory mediators such as interleukin-8 (IL-8), cyclooxygenase-2 (COX-2), macophage inhibitory cytokine 1 (MIC-1) were assessed in the human alveolar epithelial cells. Exposure to P. fluorescens and its recombinant bacteria suppressed cellular viability in the A549 epithelial cells and pro-inflammatory cytokine interleukin-8 production. However, pro-inflammatory prostaglandin-producing COX-2 protein was not altered by P. fluorescens though its mRNA was slightly elevated. As the inhibitory cytokine for the pro-inflammatory mediators, MIC-1 expression was monitored in A549 cells. MIC-1 gene induction was not significantly enhanced but the protein processing was changed by exposure to P. fluorescens. Pro-protein form of MIC-1 (${\sim}40\;kD$) was cleaved into active form mature MIC-1 (${\sim}15\;kD$) and propeptide (${\sim}28\;kD$) by the bacteria exposure. MIC-1 activation can contribute to the suppression of cellular viability by P. fluorescens and can retard IL-8-induced monocyte recruitment. However, sustained activation of MIC-1 can mediate the tissue injury by P. fluorescens exposure.

본 연구는 재조합 단백질 생산에 흔히 사용되는 P. fluorescens에 대한 공기 중 노출 시 염증작용이 의심되는 기전을 분석하기 위하여 인체 폐포 상피세포에서의 염증성 인자들 특히 IL-8, COX-2, MIC-1의 발현을 분석하고자 하였다. 균주 P. fluorescens에 대한 인체 폐포 상피세포 A549에서의 세포증식 억제효과를 밝혔고, 상피세포의 염증성 사이토카인으로서 대표적인 인터루킨 8의 발현에 대해서 분석결과. 균주 P. fluorescens및 재조합 균주에 의해 IL-8의 분비가 진핵세포대비 세균세포숫자 의존적으로 생성량이 증대되었다. 또한 점막 상피세포의 염증성 프로스타글란딘 생성에서 핵심적인 역할을 하는 cyclooxygenase-2 (COX-2)에 대해서 P. fluorescens 는 COX-2의 mRNA 발현이 소량 증진되었으나 실제 단백질의 양 및 전사의 변화는 없었다. 일반적으로 단핵구의 염증성 사이토카인 생성에 대하여 억제성 작용을 하는 macrophage inhibitory cytokine 1 (MIC-1)의 발현에 대해서 측정결과 유전자의 발현이 증대되는 효과는 미비하였으나, MIC-1 단백질의 processing에서 propeptide (${\sim}28\;kD$) 및 mature MIC-1 (${\sim}15\;kD$)로 분해 되어 MIC-1의 단백질 활성화가 증대되었다. 본 연구에서 보이는 MIC-1은 P. fluorescens에 의한 세포 생존율 억제작용에도 기여할 것으로 예측되며, IL-8에 의한 염증구의 recruitment 대한 억제작용이 예상되나, MIC-1의 활성이 과도하고 지속적으로 나타날 경우 조직의 손상도 가능성도 있다.

Keywords

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