IL-1β在大鼠角膜上皮抗曲霉菌感染固有免疫阶段的表达及其与Dectin-1受体的关系

发布时间：2018-06-02 22:41

本文选题：IL-1β + 角膜上皮　；参考：《青岛大学》2012年硕士论文

【摘要】：目的：观察IL-1β在大鼠角膜上皮抗曲霉菌感染固有免疫阶段的表达,阻断Dectin-1受体后,观察固有免疫阶段Dectin-1受体与IL-1β的关系。方法：健康的Wistar大鼠90只,雌雄不限,在刮除上皮的大鼠角膜涂布以烟曲霉菌菌丝并覆盖自制角膜接触镜,建立角膜真菌感染模型。随机分5组,A组6只为空白对照组；B组30只为真菌感染模型组,D组12只为模型阻断组,C组30只、E组12只分别为损伤对照组。均取右眼为实验眼,B组建立真菌性角膜炎模型,损伤对照组覆盖角膜接触镜并滴入PBS液；D组给予Dectin-1受体抗体后建立模型,左眼给予PBS后建立模型作为对照。建模后β、C组分别于4h,8h,16h,24h,D、E组于24h取眼球角膜上皮,运用免疫组织化学和RT-PCR技术检测角膜上皮中Dectin-1受体和IL-1β的表达。结果：IL-1β在空白对照组和条件对照组的角膜上皮呈微量表达,在真菌性角膜炎模型建立后IL-1β在角膜上皮中的表达量随病变进展逐渐增高,建模后16h至24h缓慢增高,各时间点1L-1β表达量的差异有统计学意义(P0.01)。Dectin-1受体在空白对照组角膜上皮呈基础表达,在真菌性角膜炎模型建立后Dectin-1受体表达量明显增多,差异具有显著性(P0.01)。阻断Dectin-1受体后真菌性角膜炎建模24h的IL-1β表达量明显低于未阻断Dectin-1受体组,差异具有显著性(P0.01)。结论：1.显微镜下观察大鼠角膜水肿,浑浊,溃疡形成,HE染色见角膜中性粒细胞浸润,角膜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模成功。2.IL-1β表达在真菌性角膜炎上皮中,其表达量随病变发展逐渐增高,提示IL-1β参与真菌性角膜炎的发生发展,是真菌性角膜炎病变中的敏感炎性因子。3. Dectin-1受体在空白对照组呈基础表达,在建模后Dectin-1表达量增多,Dectin-1抗体封闭Dectin-1受体后,能显著减少角膜上皮中IL-1β的量。这提示Dectin-1参与角膜上皮对烟曲霉菌菌丝的识别和反应。
[Abstract]:Aim: to observe the expression of IL-1 尾 in the innate immune phase of rat corneal epithelial anti-Aspergillus infection, and to investigate the relationship between Dectin-1 receptor and IL-1 尾 after blocking the Dectin-1 receptor. Methods: 90 healthy Wistar rats, male and female, were coated with Aspergillus fumigatus mycelium and self-made contact lens to establish the model of corneal fungal infection. Five groups were randomly divided into 5 groups: control group (n = 6), group B (n = 30), model group B (n = 30), group D (n = 12), model group C (n = 30), group E (n = 12), injury control group (n = 12). The model of fungal keratitis was established in the right eye as experimental eye group B, and the model was established in group D after Dectin-1 receptor antibody was given to the injured control group after covering the cornea contact lens and dripping into PBS solution. The model was established after PBS was given to the left eye as control group. After modeling, the corneal epithelium of group 尾 C was collected at 4 h, 8 h, 16 h and 24 h, respectively. The expression of Dectin-1 receptor and IL-1 尾 in corneal epithelium was detected by immunohistochemistry and RT-PCR technique. Results the expression of IL-1 尾 in the corneal epithelium of the control group and the control group was slight. After the establishment of the fungal keratitis model, the expression of IL-1 尾 in the corneal epithelium increased gradually with the progression of the lesion, and increased slowly from 16 hours to 24 hours after the establishment of the model. There were significant differences in the expression of 1L-1 尾 between different time points. The expression of 1L-1 尾 in the corneal epithelium of the blank control group showed a basic expression. After the establishment of the fungal keratitis model, the expression of Dectin-1 尾 increased obviously, and the difference was significant (P 0.01). The expression of IL-1 尾 in fungal keratitis model 24 h after blocking Dectin-1 receptor was significantly lower than that in unblocked Dectin-1 receptor group (P 0.01). Conclusion 1. Under microscope, corneal edema, turbidity, ulcer formation and neutrophil infiltration were observed in rats. Colony culture of corneal tissue scraping confirmed that the rat model of fungal keratitis was successfully established. 2. The expression of IL-1 尾 in the epithelium of fungal keratitis increased with the development of the lesion, suggesting that IL-1 尾 was involved in the occurrence and development of fungal keratitis. It is a sensitive inflammatory factor in fungal keratitis. The expression of Dectin-1 receptor was basic in blank control group. After modeling, the increase of Dectin-1 expression and the blocking of Dectin-1 receptor by Dectin-1 antibody could significantly reduce the amount of IL-1 尾 in corneal epithelium. This suggests that Dectin-1 is involved in the recognition and response of corneal epithelium to Aspergillus fumigatus hyphae.
【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】：R772.2

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