大鼠中耳炎细菌生物膜模型的建立及其组织形态学的观察

发布时间：2019-01-09 07:28

【摘要】： 急性中耳炎(acute otitis media,AOM)是儿童期发病仅次于上呼吸道感染的最常见的感染性疾病,其中最困扰临床的是复发性AOM和顽固性AOM,据统计,约20%-30%的儿童罹患复发性AOM,且这一数据还在逐年增加。频繁的中耳感染不仅会引起患儿鼓室硬化、粘连性中耳炎以及中耳胆脂瘤,从而导致患儿听力进行性下降,甚至会影响到语言的发育。然而,其确切发病机制、抗生素耐药机理以及反复发作的原因等至今仍不清楚。自2001年Post等首次在分泌性中耳炎的发病机制中提出细菌生物膜的致病理论以来,细菌生物膜与中耳炎的关系迅速成为学者们关注的焦点,但目前文献多侧重于研究细菌生物膜和分泌性中耳炎的关系。2009年,Reid等和Hoa等分别在不同的粟鼠AOM动物模型上观察到有细菌生物膜的存在,但细菌生物膜在AOM中的形成特点、是否参与AOM的发病过程以及临床意义等,仍有待深入研究。基于此,我们设计了本课题。研究分两个部分进行,在第一部分研究中,我们探讨了通过听泡穿刺、将肺炎链球菌注入到大鼠中耳腔建立大鼠AOM模型的可行性,并通过观察其病理变化和病程,探讨了该模型的应用价值,以期为后续的深入研究提供基础资料。研究证实,将50ul肺炎链球菌悬液(1×108CFU/ml)经听泡穿刺注入到大鼠中耳腔建立了急性中耳炎的大鼠模型,成功率100%,是研究人类中耳炎较为理想的动物模型,不仅可用于急性化脓性中耳炎的研究,也可用于急性分泌性中耳炎的研究。在成功建立了急性中耳炎大鼠模型的基础上,我们进行了第二部分的研究,采用扫描电镜并结合免疫荧光原位标记技术、激光扫描共聚焦显微镜来研究细菌生物膜在急性中耳炎大鼠中耳腔的形成及其临床意义,并探讨大鼠AOM动物模型用于细菌生物膜研究的可行性。研究发现,细菌生物膜在中耳急性感染的早期即已开始形成,并可能成为日后炎症反复发作或迁延不愈的原因,早期及时应用抗生物膜药物有助于感染的彻底根除,本研究建立的大鼠急性中耳炎模型是研究细菌生物膜较为理想的动物模型。
[Abstract]:Acute otitis media (acute otitis media,AOM) is the most common infectious disease in childhood, second only to upper respiratory tract infection, among which recurrent AOM and intractable AOM, are the most troubling disease in children. According to statistics, about 20% to 30% of children suffer from recurrent AOM,. And this figure is still increasing year by year. Frequent middle ear infections not only cause tympanic sclerosis, adhesive otitis media and cholesteatoma of the middle ear, but also lead to progressive decline of hearing and even affect the development of language. However, the exact mechanism of its pathogenesis, antibiotic resistance and the causes of repeated attacks are still unclear. Since 2001, when Post et al first put forward the pathogenic theory of bacterial biofilm in the pathogenesis of secretory otitis media, the relationship between bacterial biofilm and otitis media has become the focus of attention. In 2009, Reid et al and Hoa et al observed the presence of bacterial biofilm in different AOM animal models of millet, but the formation characteristics of bacterial biofilm in AOM. Whether or not to participate in the pathogenesis and clinical significance of AOM remains to be further studied. Based on this, we designed the subject. In the first part of the study, we discussed the feasibility of injecting Streptococcus pneumoniae into the middle ear cavity of rats to establish AOM model, and observed its pathological changes and course of disease. The application value of the model is discussed in order to provide basic data for further research. It was proved that the rat model of acute otitis media was established by injecting 50ul pneumoniae suspension (1 脳 108CFU/ml) into the middle ear cavity of rats through auditory bubble puncture, and the success rate was 100%, which was an ideal animal model for the study of human otitis media. It can be used not only in the study of acute suppurative otitis media, but also in the study of acute secretory otitis media. On the basis of the successful establishment of the rat model of acute otitis media, we carried out the second part of the study, using scanning electron microscope (SEM) and immunofluorescence in situ labeling technique. The formation and clinical significance of bacterial biofilm in the middle ear cavity of rats with acute otitis media were studied by laser scanning confocal microscope and the feasibility of using the animal model of AOM in the study of bacterial biofilm was discussed. The study found that bacterial biofilm had begun to form at the early stage of acute infection in the middle ear, and might become the cause of repeated or prolonged inflammation in the future. The early and timely application of anti-biofilm drugs was helpful to the complete eradication of infection. The rat model of acute otitis media is an ideal animal model to study bacterial biofilm.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R764

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