Dectin-1在大鼠真菌性角膜炎固有免疫阶段的作用

发布时间：2018-02-03 09:19

本文关键词： Dectin-1 Raf-1 PLCγ-2 固有免疫角膜上皮　出处：《青岛大学》2013年硕士论文　论文类型：学位论文

【摘要】：目的：观察树突状细胞相关性c型凝集素1(Dendritic cell-associated C-type lectin-1,dectin-1)受体在真菌性角膜炎角膜上皮中的表达,探索Dectin-1的信号通路,探讨角膜真菌感染中角膜上皮表达的Dectin-1在真菌性角膜炎固有免疫阶段的作用。方法：健康Wistar大鼠54只,随机分成空白对照组(n=6)、真菌性角膜炎组(n=48)。根据是否应用Laminarin预处理,再将真菌性角膜炎组分为阻断组(n=24)和真菌阳性对照组(n=24)；阻断组用Laminarin点眼处理2h,后与真菌性角膜炎组同时制备真菌性角膜炎动物模型,分别于模型制备24h后处死大鼠,摘取眼球免疫组织化学染色法检测ccl2、ccl3、cxcl1、cxcl2、IL-1β、TNF-α、IL-6、IL-10八种因子蛋白表达。实时荧光定量PCR法检测真菌性角膜炎模型制备4h、8h、16h、24h后角膜上皮中以上八种因子mRNA表达水平。采用t检验进行组间数据比较。结果：时荧光定量PCR法检测结果发现,空白组8种因子均有表达,真菌刺激4h后il-1β、il-6、ccl2、cxcl1、cxcl2表达增高,差异有统计学意义(p0.05),真菌刺激8h后tnf-α/ccl3表达较空白组增高,差异有统计学意义(p0.05)；真菌刺激4h,阻断组较真菌组il-1β、il-6、ccl2、cxcl1、cxcl2表达下降,差异有统计学意义(p0.05),其余3种因子差异无统计学意义,真菌刺激8h,阻断组i1-6和cxcl1较真菌组表达仍下降,差异有统计学意义(p0.05)。结论：Dectin-1在角膜上皮细胞中表达,烟曲霉菌刺激可引起Dectin-1表达升高,Dectin-1可能通过Raf-1信号通路引起炎症因子的表达,在大鼠真菌性角膜炎固有免疫早期可能通过趋化因子il-1β、il-6、ccl2、cxcl1、cxcl2募集中性粒细胞和巨噬细胞参与机体抗真菌免疫。
[Abstract]:Objective: to observe the dendritic cell-related c type lectin 1 cell-associated C-type lectin-1. Expression of dectin-1) receptor in corneal epithelium of fungal keratitis to explore the signal pathway of Dectin-1. To investigate the role of Dectin-1 expressed in corneal epithelium in the innate immune stage of fungal keratitis. Methods: 54 healthy Wistar rats were randomly divided into control group (n = 6) and fungal keratitis group (n = 48). The fungal keratitis group was divided into two groups: the blocking group and the fungal positive control group. The rats in the blocking group were treated with Laminarin for 2 hours, and then the fungal keratitis model was established simultaneously with the fungal keratitis group. The rats were killed 24 hours after the establishment of the model. Immunohistochemical staining was used to detect the expression of ccl2ccl3cxcl1cxcl2ccl1- 尾 and TNF- 伪 IL-6. The expression of eight kinds of IL-10 protein was detected by real-time fluorescence quantitative PCR. The model of fungal keratitis was established for 4 h and 8 h for 16 h. After 24 hours, the mRNA expression level of the above eight factors in corneal epithelium was compared by t test. Results: the results of time-fluorescence quantitative PCR assay showed that all the eight factors were expressed in blank group, and il-1 尾 -ccl2cxcl1 was stimulated by fungi for 4 h. The expression of tnf- 伪 / ccl3 was significantly higher than that of the control group after 8 h of fungal stimulation, and the difference was statistically significant (P 0.05). The expression of cxcl2cxcl1cxcl2 in the blocking group was significantly lower than that in the fungal group (p0.05). There was no significant difference in the other three factors. The expression of i1-6 and cxcl1 in the blocking group was still lower than that in the fungal group after 8 h of fungal stimulation, and the difference was statistically significant (P 0.05). ConclusionThe expression of Dectin-1 in corneal epithelial cells was induced by the stimulation of aspergillus fumigatus, and the expression of Dectin-1 was increased by the stimulation of aspergillus fumigatus. Dectin-1 may induce the expression of inflammatory factors through the Raf-1 signaling pathway, and in the early stage of the innate immunity of rat fungal keratitis, it may be mediated by the chemokine il-1 尾 -il-6 ccl2. The recruitment of neutrophils and macrophages involved in antifungal immunity.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R772.21

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