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目的 TLR3介导的天然免疫通路是抗感染重要的天然免疫机制,本文研究鼻感染烟曲霉菌是否能激活小鼠肺组织TLR3天然免疫信号通路。方法小鼠分正常组和感染组,感染组经鼻感染烟曲霉菌孢子,正常组经鼻滴入等量的生理盐水,8 h、24 h和48 h后处死小鼠,分离肺组织,取100 mg肺组织RT-PCR检测TLR3、TRIF及IFN-βmRNA水平、Western blot检测胞核IRF3的含量。取24 h、48 h和72 h小鼠肺组织制作病理切片,100 mg肺组织碾碎接种蔡氏培养基平板以检测肺组织烟曲霉菌载荷。结果感染组小鼠肺组织烟曲霉菌负荷72 h时显著下降,正常组烟曲霉菌培养阴性;正常组小鼠肺组织结构正常,感染组小鼠24 h时肺组织可见严重的炎症反应及出血,72 h时肺组织炎症反应减轻;RT-PCR及Western blot结果显示:感染组小鼠肺组织各时相点TLR3 mRNA、TRIF mRNA、IRF3和IFN-βmRNA的表达量均显著高于正常对照组(P<0.05),均在24 h达到最高峰(P<0.05)。结论烟曲霉菌感染的小鼠肺组织中TLR3信号通路得以激活,其介导的天然免疫对感染烟曲霉菌的清除和保护小鼠肺组织起了一定的作用。
Objective TLR3-mediated innate immune pathway is an important natural immune mechanism against infection. In this paper, whether nasal infection with Aspergillus fumigatus could activate TLR3 innate immune signaling pathway in mouse lung tissue was investigated. Methods The mice were divided into normal group and infected group. Infection group was infected with Aspergillus fumigatus spores by nasal injection. The rats in normal group were injected with the same amount of normal saline. The mice were sacrificed at 8, 24 and 48 hours. The levels of TLR3, TRIF and IFN-βmRNA were detected by RT-PCR in 100 mg lung tissue. The content of IRF3 in nucleus was detected by Western blot. Pathological sections of lungs of mice were made at 24 h, 48 h and 72 h, and 100 mg of lung tissue was inoculated with Chua’s medium to detect Aspergillus fumigatus load in lung tissue. Results Aspergillus fumigatus in the lungs of infected mice decreased significantly at 72 h, and the culture of Aspergillus fumigatus in normal group was negative. The normal lung tissues of mice in normal group were normal, and severe inflammatory reaction and hemorrhage , And the inflammatory response of lung tissue was relieved at 72 h. The expression of TLR3 mRNA, TRIF mRNA, IRF3 and IFN-βmRNA in the lung tissues of the infected mice at each time point was significantly higher than that of the normal control group (P <0.05), all reaching the peak at 24 h (P <0.05). Conclusion TLR3 signaling pathway is activated in the lungs of Aspergillus fumigatus-infected mice. The innate immunity mediated by it plays a role in clearance of Aspergillus fumigatus and protection of lung tissue of mice.