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  本文關(guān)鍵詞： Dectin-1 Raf-1 PLCγ-2 固有免疫 角膜上皮　出處：《青島大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：觀察樹突狀細(xì)胞相關(guān)性c型凝集素1(Dendritic cell-associated C-type lectin-1,dectin-1)受體在真菌性角膜炎角膜上皮中的表達(dá),探索Dectin-1的信號通路,探討角膜真菌感染中角膜上皮表達(dá)的Dectin-1在真菌性角膜炎固有免疫階段的作用。 方法：健康Wistar大鼠54只,隨機(jī)分成空白對照組(n=6)、真菌性角膜炎組(n=48)。根據(jù)是否應(yīng)用Laminarin預(yù)處理,再將真菌性角膜炎組分為阻斷組(n=24)和真菌陽性對照組(n=24)；阻斷組用Laminarin點(diǎn)眼處理2h,后與真菌性角膜炎組同時制備真菌性角膜炎動物模型,分別于模型制備24h后處死大鼠,摘取眼球免疫組織化學(xué)染色法檢測ccl2、ccl3、cxcl1、cxcl2、IL-1β、TNF-α、IL-6、IL-10八種因子蛋白表達(dá)。實(shí)時熒光定量PCR法檢測真菌性角膜炎模型制備4h、8h、16h、24h后角膜上皮中以上八種因子mRNA表達(dá)水平。采用t檢驗(yàn)進(jìn)行組間數(shù)據(jù)比較。 結(jié)果：時熒光定量PCR法檢測結(jié)果發(fā)現(xiàn),空白組8種因子均有表達(dá),真菌刺激4h后il-1β、il-6、ccl2、cxcl1、cxcl2表達(dá)增高,差異有統(tǒng)計(jì)學(xué)意義(p0.05),真菌刺激8h后tnf-α/ccl3表達(dá)較空白組增高,差異有統(tǒng)計(jì)學(xué)意義(p0.05)；真菌刺激4h,阻斷組較真菌組il-1β、il-6、ccl2、cxcl1、cxcl2表達(dá)下降,差異有統(tǒng)計(jì)學(xué)意義(p0.05),其余3種因子差異無統(tǒng)計(jì)學(xué)意義,真菌刺激8h,阻斷組i1-6和cxcl1較真菌組表達(dá)仍下降,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。 結(jié)論：Dectin-1在角膜上皮細(xì)胞中表達(dá),煙曲霉菌刺激可引起Dectin-1表達(dá)升高,Dectin-1可能通過Raf-1信號通路引起炎癥因子的表達(dá),在大鼠真菌性角膜炎固有免疫早期可能通過趨化因子il-1β、il-6、ccl2、cxcl1、cxcl2募集中性粒細(xì)胞和巨噬細(xì)胞參與機(jī)體抗真菌免疫。
[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- 偽 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- 偽 / 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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R772.21

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本文編號：1487001