肺是維持生命的重要器官，使用呼吸器的目的主要是協助病人渡過急性呼吸衰竭，但長時間使用可能對肺部造成二次傷害並增加呼吸器相關性肺炎(ventilator-associated pneumonia, VAP)的發生，嚴重時甚至會導致死亡。呼吸器會降低肺泡巨噬細胞的抗菌能力而增加呼吸器相關性肺炎的發生。近來許多研究指出鮑氏不動桿菌(A. baumannii)是加護病房中病患因使用呼吸器發生肺炎時常見的致病菌，且造成的感染因對抗生素具抗性而難以治療。因此，為探討鮑氏不動桿菌造成肺部損傷的機制，將鮑氏不動桿菌以鼻腔注射方式注入C57BL/6 (WT)和JNK1-/-小鼠，並於48小時後連接呼吸器3小時再進行肺部損傷情形分析。結果顯示，給予鮑氏不動桿菌再連接呼吸器時，WT小鼠肺部的嗜中性白血球浸潤程度、肺泡沖提液的一氧化氮含量皆明顯增加，但在體外將巨噬細胞以鮑氏不動桿菌刺激的上清液中TNF-α含量明顯少於綠膿桿菌刺激的組別，且在體外對鮑氏不動桿菌的抗菌能力也較差，此結果極可能是造成肺部損傷的主因。而當給予鮑氏不動桿菌再連接呼吸器時，JNK1-/-小鼠的肺部發炎及損傷均減緩，且其巨噬細胞受鮑氏不動桿菌刺激後上清液中TNF-α含量低於WT小鼠，而當IKK△mye小鼠的巨噬細胞受鮑氏不動桿菌刺激後，上清液中TNF-α的含量則明顯下降。綜合以上結果，本研究模式中，鮑氏不動桿菌及呼吸器會促進肺部NF-κB及JNK訊息傳遞路徑的活化，進而增加肺部損傷的程度。

Ventilator-associated pneumonia (VAP) continues to be a major cause of high morbidity and mortality for patients using ventilators. Patients in ICUs often receive broad-spectrum antibiotic treatment and multidrug resistant Pseudomonas aeruginosa and Acinetobacter baumannii were the most common pathogens causing VAP. When lung is stimulated, lung-resident monocytes/ macrophages are recruited and release inflammatory cytokines/chemokines which induce lung injury. Alveolar macrophages (AMs) are in most frequent contact with external stimuli which can initiate phagocytosis to clear infection and regulate the secretion of cytokines/chemokines. Previous studies with a mouse model showed that VAP caused by P. aeruginosa involved TNF-α production through NF-κB activation in AMs and JNK signaling in lungs. In current study, C57BL/6 mice intranasally instilled with A. baumannii showed a significant increase of neutrophil infiltration into lungs after ventilation, and the NO level in bronchoalveolar lavage fluid (BALF) collected from mice instilled with A. baumannii was higher than instilled with P. aeruginosa. When JNK1-/- mice receiving the instillation of A. baumannii, the level of MPO activity, total cells in BALF, nitrite in BALF and nitrite in serum were all lower when compared to WT mice. These results indicated a lower inflammation level and lung damage in JNK1-/- mice. However, the killing activity of AMs against A. baumannii was lower than P. aeruginosa whether with or without ventilation. The decreased killing activity was parallel with decreased TNF-α production by AMs when challenged with A. baumannii. These results suggest that the pathogenesis mechanism of VAP caused by A. baumannii was related to the activation of NF-κB and JNK signaling pathway.

論文審定書………..……………………………………………………...i
論文授權書…………..…………………………………………………..ii
致謝……………...…..…………………………………………………..iii
Abstract in Chinese……………………………………………………....iv
Abstract in English………………………………………………………..v
List of Abbreviations……………………………………………………vii
Introduction……………………………………………………………….1
Materials and Methods……………………………………………...…..10
Results…………………………………………………………...……....18
Discussion………………………………..……………………….…......27
Figures…………………………………………………………………...36
Tables...………………………………………………...………………..58
Appendix…………………………………………………………….......62
References……………………………………………………………….64

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