IL-6与IL-10在大鼠真菌性角膜炎固有免疫阶段角膜上皮组织中的表达

发布时间：2018-04-30 13:25

本文选题：烟曲霉菌 + 角膜上皮细胞　；参考：《青岛大学》2012年硕士论文

【摘要】：目的观察白细胞介素6(interleukin-6,-6)和白细胞介素10(interleukin-0,IL-10)在大鼠烟曲霉菌性角膜炎固有免疫阶段角膜上皮组织中的表达并分析其表达量变化的原因。方法健康Wi star大鼠72只,雌雄不限,在刮除中央区角膜上皮的大鼠角膜上涂以烟曲霉菌标准菌落并覆盖自制角膜接触镜,建立大鼠烟曲霉菌性角膜炎感染模型。大鼠随机分为2部分,用于免疫组织化学实验的A1、B1、C1组(每组各6只大鼠)和用于RT-PCR实验的A2、B2、C2(A2组6只、B2、C2组24只)：A1、A2组为空白对照组,不予任何处理；B1、B2组为损伤对照组,建立模型,仅刮除中央区角膜上皮后覆盖角膜接触镜,不涂抹真菌；C1、C2组为真菌模型组,建立真菌模型。成功建模后4h打开眼睑,并于造模后第4h、8h、16h、24h处死大鼠,取角膜上皮组织。HE染色观察角膜组织病理变化并检测真菌生长,运用免疫组织化学和RT-PCR技术检测各组不同时间点IL-6与IL-10的表达情况。结果1.组织病理学观察见真菌性角膜炎病灶内有炎症细胞浸润,HE染色检查在角膜基质层可见菌丝及孢子。2.IL-6在空白对照眼角膜基本不表达或极微量表达；真菌实验组在接种真菌后4h开始出现IL-6的表达,并逐渐增高,在第24h表达量升高显著,各时间点比较差异具有显著性(P0.01)。3.IL-10在空白对照组角膜基本不表达或极微量表达；真菌实验组在真菌刺激后4h开始出现IL-10的表达,并逐渐增高,在24h表达量升高显著,各时间点比较差异有显著性((P0.01)。结论1.10%氢氧化钾涂片及组织刮取物行菌落培养证实大鼠真菌性角膜炎动物模型建立成功。2.IL-6在烟曲霉菌性角膜炎角膜上皮组织中早期即开始表达,表达量逐渐增高,其表达量的增高与角膜炎症程度呈正相关,在大鼠烟曲霉菌性角膜炎病程中是一个敏感的炎症因子,能反映局部的炎症反应程度。3.IL-10表达于真菌性角膜炎角膜上皮组织中,烟曲霉菌菌丝刺激后表达升高,24h升高显著,提示IL-10可能参与了烟曲霉菌性角膜炎的发生与发展,作为一种保护性细胞因子通过抗炎抗免疫反应也参与了角膜组织修复和抗损伤过程。
[Abstract]:Objective to observe the expression of interleukin-6 (IL-6) and interleukin-0 (IL-10) in the corneal epithelium of rats with tobacco-curving fungal keratitis and to analyze the causes of the changes in the expression of interleukin-6 (IL-6) and interleukin-0 (IL-10) in the cornea of rats with tobacco-curving fungal keratitis. Methods 72 healthy Wi star rats, male and female, were smeared with standard colony of Aspergillus fumigatus and covered with self-made contact lens on the cornea of rats with central corneal epithelium. Rats were randomly divided into two groups: group A _ 1B _ 1C _ 1 for immunohistochemistry (n = 6 for each group) and group A _ 2 B _ 2C _ 2 for RT-PCR test. 24 rats in B _ 2C _ 2 group were served as blank control group, and B _ 1B _ 2 group was used as the injury control group without any treatment, and the model was established in B _ 1B _ 2 group. Only the corneal epithelium in the central area was scraped over the contact lens, and the fungal model was established in the group C _ (1) C _ (2) without smear. The eyelids were opened at 4 h after successful modeling, and the rats were killed at 4 h, 8 h, 16 h and 24 h. The corneal epithelium was stained with HE to observe the pathological changes of corneal tissue and to detect the growth of fungi. Immunohistochemical and RT-PCR techniques were used to detect the expression of IL-6 and IL-10 at different time points. Result 1. Histopathological observation showed that there were inflammatory cells infiltrating in the focus of fungal keratitis. The mycelium and spores of mycelium and spore. 2. IL-6 were not expressed in the cornea of the blank control group. The expression of IL-6 began to appear at 4 hours after inoculation, and increased gradually in the fungal experimental group, and the expression level increased significantly at 24 h after inoculation, and the difference at each time point was significant (P 0.01). 3. IL-10 was not expressed in the cornea of the blank control group. The expression of IL-10 began to appear at 4 h after fungal stimulation, and gradually increased, and the expression level increased significantly at 24 h, and there was a significant difference between the two groups at different time points (P 0.01). Conclusion 1.10% potassium hydroxide smear and tissue scraping for colony culture proved that the rat model of fungal keratitis was successfully established. 2. IL-6 expression began to express in the corneal epithelium of tobacillus fumigatus keratitis at the early stage, and the expression level increased gradually. The increase of its expression level was positively correlated with the degree of corneal inflammation. It was a sensitive inflammatory factor in the course of rat keratitis caused by tobacillus fumigatus, and could reflect the degree of local inflammation. IL-10 was expressed in the corneal epithelium of fungal keratitis. The increased expression of Aspergillus fumigatus mycelium increased significantly at 24 h, suggesting that IL-10 may be involved in the occurrence and development of mycelial keratitis of Aspergillus fumigatus. As a protective cytokine, it also participates in corneal tissue repair and anti-injury process through anti-inflammatory and anti-immune reaction.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R772.21

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