CAJ | 학술논문

目的：本研究利用小鼠动物模型，探讨烟曲霉感染免疫缺陷小鼠TLR2/TLR3/TLR4对树突状细胞植物血凝素-1（Dectin-1）表达的影响。通过检测TLRs和CLRs受体在不同免疫状态下的表达，揭示介导侵袭性肺曲霉菌病（IPA）肺损伤的关键受体。方法小鼠随机分成四组，A组为正常对照组；B组为环磷酰胺免疫抑制＋未接种烟曲霉菌；C组正常状态小鼠＋烟曲霉菌接种；D组为IPA感染组，免疫抑制＋烟曲霉菌接种。采用real-time PCR方法进行小鼠肺组织各个时相点的TLR2、TLR3、TLR4、Dectin-1和β-actin mRNA的表达，检测受体间的相互表达调控。结果正常状态感染组（C组N＋AF），TLR4/TLR2、Dectin-1 mRNA比正常对照组表达上调；IPA模型组（D组CP＋AF），Dectin-1和TLR4、TLR2 mRNA比正常状态感染组表达下调；但TLR3 mRNA 48 h、72 h表达没有明显不同（P＞0.05）。病理切片模型组小鼠72 h可见较严重的出血和充血；96 h时肺内有菌丝，肺泡间隔增宽，支气管壁破坏。正常状态感染烟曲霉小鼠72 h可见充血，肺泡弹性纤维破坏。结论成功建立了小鼠IPA动物模型，Dectin-1受体表达下调可能是环磷酰胺免疫抑制引起IPA的机制之一。Dectin-1的表达可能是建立在TLR2/TLR4对烟曲霉早期识别的基础上，TLR3可能起协同作用。
목적：본연구이용소서동물모형，탐토연곡매감염면역결함소서TLR2/TLR3/TLR4대수돌상세포식물혈응소-1（Dectin-1）표체적영향。통과검측TLRs화CLRs수체재불동면역상태하적표체，게시개도침습성폐곡매균병（IPA）폐손상적관건수체。방법소서수궤분성사조，A조위정상대조조；B조위배린선알면역억제＋미접충연곡매균；C조정상상태소서＋연곡매균접충；D조위IPA감염조，면역억제＋연곡매균접충。채용real-time PCR방법진행소서폐조직각개시상점적TLR2、TLR3、TLR4、Dectin-1화β-actin mRNA적표체，검측수체간적상호표체조공。결과정상상태감염조（C조N＋AF），TLR4/TLR2、Dectin-1 mRNA비정상대조조표체상조；IPA모형조（D조CP＋AF），Dectin-1화TLR4、TLR2 mRNA비정상상태감염조표체하조；단TLR3 mRNA 48 h、72 h표체몰유명현불동（P＞0.05）。병리절편모형조소서72 h가견교엄중적출혈화충혈；96 h시폐내유균사，폐포간격증관，지기관벽파배。정상상태감염연곡매소서72 h가견충혈，폐포탄성섬유파배。결론성공건립료소서IPA동물모형，Dectin-1수체표체하조가능시배린선알면역억제인기IPA적궤제지일。Dectin-1적표체가능시건립재TLR2/TLR4대연곡매조기식별적기출상，TLR3가능기협동작용。
Objective The purpose of the study is using mice animal models to explore the impact of TLR2/TLR3/TLR4 of immunosuppressed mice infected with A. fumigatus on dendritic cells phytohemagglutinin-1 (Dectin-1) expression. By detecting TLRs and CLRs receptors in different immune status expression, we reveal the key receptors-mediated lung injury IPA. Methods The mice were divided into four groups including the group of normal mice, the group of immunosuppressed mice without A. fumigatus injection, the group of normal mice with A. fumigatus injection and the group of IPA mice. The lung tissues from each group were collected for pathological analysis and detecting the expression level of TLR2, TLR3, TLR4, Dectin-1 and β-actin mRNA by real-time PCR, and detecting receptors expression and mutual regulation. Results In the immunocompetent mice infected with A. fumigatus, TLR4/TLR2 and Dectin-1 mRNA expression increased markedly compared with the normal control group. In the immunosuppressed mice infected with A. fumigatus, TLR4/TLR2 and Dectin-1 mRNA expression were inhibited markedly compared with normal mice infected with A. fumigatus, but TLR3 mRNA expression of mice injected after 48 h and 72 h did not show significant difference (P>0. 05). IPA mice infected after 72 h showed severe bleeding and congestive, and lung tissue granuloma formation;the 96 h group, showed hyphae,widened alveolar septum, tracheal epithelium cell loss, tissue necrosis. Normal mice infected by Aspergillus fumigatus after 72 h showed visible congestion and destroyed alveolar elastic fibers. Conclusion An experimental animal model of invasive pulmonary aspergillosis was established successfully. Inhibition of Dectin-1 expression may be one of the mechanisms of cyclophosphamide in the development of IPA. Dectin-1 expression may be established in the early stage of TLR2/TLR4 identification of the A. fumigatus, whereas TLR3 may play a synergistic role.