發(fā)布時(shí)間：2018-01-19 06:13

  本文關(guān)鍵詞： 急性中耳炎 小鼠 肺炎鏈球菌 動(dòng)物模型 莢膜多糖　出處：《重慶醫(yī)科大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：目的 肺炎鏈球菌是一種常見(jiàn)的革蘭陽(yáng)性條件致病菌，是引起中耳炎的主要致病菌之一。中耳炎是兒童常見(jiàn)病，反復(fù)發(fā)作易引起患兒聽(tīng)力障礙甚至缺失，延遲患兒的語(yǔ)言及智力發(fā)育。但肺炎鏈球菌中耳炎的分子病理機(jī)制至今尚不十分清楚。因此，本研究擬建立小鼠急性肺炎鏈球菌中耳炎模型，并觀察肺炎鏈球菌主要致病因子莢膜多糖在中耳炎中的作用。 方法 1．不同濃度的4型肺炎鏈球菌（TIGR4）懸液經(jīng)鼓膜穿刺接種于C57BL/6J小鼠中耳腔中，建立小鼠急性肺炎鏈球菌中耳炎模型。觀察小鼠體重變化及發(fā)病情況，取心臟血及中耳灌洗液進(jìn)行細(xì)菌培養(yǎng)，中耳灌洗液進(jìn)行細(xì)胞計(jì)數(shù)、流式細(xì)胞分類計(jì)數(shù)和細(xì)胞涂片瑞氏染色，，并測(cè)定中耳灌洗液上清和血清中細(xì)胞因子含量，小鼠的中耳組織進(jìn)行組織切片HE染色。 2．TIGR4和莢膜缺陷菌TIGR4Δcps分別經(jīng)鼓膜穿刺接種于中耳腔中，通過(guò)觀察小鼠體重變化及發(fā)病情況、小鼠中耳灌洗液細(xì)菌培養(yǎng)、細(xì)胞計(jì)數(shù)、細(xì)胞涂片瑞氏染色和流式分類計(jì)數(shù)及細(xì)胞因子水平的差異來(lái)研究肺炎鏈球菌莢膜多糖在中耳炎中的作用。 結(jié)果 1．小鼠中耳腔接種肺炎鏈球菌后，小鼠體重下降，呈劑量依賴。中耳粘膜上皮細(xì)胞壞死脫落，粘膜增厚，中耳腔內(nèi)大量炎癥細(xì)胞浸潤(rùn)，中耳骨壁骨質(zhì)破壞。中耳腔內(nèi)中性粒細(xì)胞的募集時(shí)相與細(xì)菌的清除時(shí)相呈相關(guān)性，中耳灌洗液上清中IL-6和TNF-α顯著升高。 2．與野生型TIGR4相比，TIGR4Δcps在中耳腔中的定植顯著減少，清除加快，募集的炎癥細(xì)胞數(shù)量以及IL-6和TNF-α水平顯著低于野生組。 結(jié)論 本研究成功建立了小鼠急性肺炎鏈球菌中耳炎模型，中耳微環(huán)境中炎癥細(xì)胞和炎癥因子的募集增加。同時(shí)，研究發(fā)現(xiàn)在肺炎鏈球菌中耳炎中，莢膜多糖具有促進(jìn)細(xì)菌定植、募集炎癥細(xì)胞和誘導(dǎo)炎癥因子釋放等重要作用。
[Abstract]:Purpose Streptococcus pneumoniae is a common gram-positive conditional pathogen, which is one of the main pathogens causing otitis media. Otitis media is a common disease in children. The molecular pathological mechanism of Streptococcus pneumoniae otitis media is still unclear. Therefore, this study intends to establish a mouse model of acute streptococcus pneumoniae otitis media. To observe the role of capsular polysaccharide, the main pathogenic factor of Streptococcus pneumoniae, in otitis media. Method 1. Different concentrations of Streptococcus pneumoniae TIGR4 suspension were inoculated into the middle ear cavity of C57BL / 6J mice by tympanic membrane puncture. A mouse model of acute streptococcus pneumoniae otitis media was established. The changes of body weight and the incidence of the disease were observed. The heart blood and the middle ear lavage fluid were taken for bacterial culture and the middle ear lavage fluid was counted. The levels of cytokines in the supernatant of middle ear lavage fluid and serum were determined. The middle ear tissue of mice was stained with HE in tissue sections. 2. TIGR4 and TIGR4 螖 cps were inoculated into the middle ear cavity by tympanic membrane puncture. To study the role of streptococcus pneumoniae capsule polysaccharides in otitis media, cell count, cell smear staining, flow classification count and cytokine level were different in order to study the role of streptococcus pneumoniae capsule polysaccharides in otitis media. Results 1. After inoculation of Streptococcus pneumoniae in the middle ear cavity of mice, the weight of the mice decreased in a dose-dependent manner. The epithelial cells of the middle ear were necrotic, the mucosa thickened, and a large number of inflammatory cells infiltrated in the middle ear cavity. There was a correlation between the recruitment phase of neutrophils and the clearance of bacteria in the middle ear cavity. The IL-6 and TNF- 偽 in the supernatant of the middle ear lavage fluid increased significantly. 2. Compared with wild-type TIGR4, the colonization of TIGR4 螖 cps in the middle ear cavity was significantly reduced and the clearance was accelerated. The number of inflammatory cells recruited and the levels of IL-6 and TNF- 偽 were significantly lower than those in the wild group. Conclusion In this study, the mouse model of acute streptococcus pneumoniae otitis media was successfully established, and the recruitment of inflammatory cells and inflammatory factors in the middle ear microenvironment was increased. At the same time, it was found that in Streptococcus pneumoniae otitis media. Capsule polysaccharide plays an important role in promoting bacterial colonization, recruiting inflammatory cells and inducing the release of inflammatory factors.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R764.21;R-332

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本文編號(hào)：1442978