發(fā)布時(shí)間：2018-12-25 14:02

【摘要】：目的通過建立真菌性角膜炎小鼠模型，觀察真菌性角膜炎的發(fā)生發(fā)展，探索TH17細(xì)胞因子在真菌性角膜炎中的表達(dá)情況，初步探討IL-17及TH17細(xì)胞在真菌性角膜炎免疫學(xué)中的作用。 方法清潔級BALB/c小鼠隨機(jī)分成兩組，其中一組為真菌感染組，小鼠右眼采用角膜表層鏡片法建立茄病鐮刀菌性角膜炎模型；另一組為損傷對照組，以PBS代替真菌菌液采取與感染組相同方法處理小鼠右眼。分別于感染后第1、3、5、7天，裂隙燈顯微鏡觀察角膜炎特點(diǎn)并行臨床評分；HE染色觀察角膜病理變化；實(shí)時(shí)熒光定量PCR檢測角膜中Th17細(xì)胞因子（IL-17）及其特異性轉(zhuǎn)錄因子RORγt的mRNA基因的表達(dá)變化；酶聯(lián)免疫吸附試驗(yàn)檢測IL-17因子蛋白的表達(dá)變化。 結(jié)果1.病變特征 所有小鼠真菌性角膜炎模型建立成功，真菌感染早期呈全角膜的彌漫性炎癥反應(yīng)，，浸潤混濁進(jìn)行性加重，感染后3天角膜灰白色霧狀混濁，５天時(shí)病變典型，角膜周邊新生血管長入，中央形成潰瘍，第７天后病灶趨向好轉(zhuǎn)，總病程持續(xù)約12天。臨床評分各時(shí)間點(diǎn)有統(tǒng)計(jì)學(xué)差異（P＜0.05）。 2.病理學(xué)檢查 HE染色可見早期角膜基質(zhì)水腫，感染后第5天角膜基質(zhì)紊亂多形粒細(xì)胞浸潤明顯增多，前房內(nèi)可見滲出，晚期角膜基質(zhì)中可見新生血管及管內(nèi)紅細(xì)胞，潰瘍被瘢痕組織取代。 3.Real time PCR與ELISA檢測結(jié)果 Thl7細(xì)胞因子（IL-17）和Th17細(xì)胞特異性轉(zhuǎn)錄因子（ROR γt）基因mRNA在真菌感染角膜后均出現(xiàn)表達(dá)，于第3、5、7天顯著高于損傷對照組（P＜0.01），且組內(nèi)比較5天時(shí)表達(dá)量最高（P＜0.01）； IL-17因子蛋白感染后各時(shí)間點(diǎn)表達(dá)量顯著高于損傷對照組（P＜0.01）。 結(jié)論新型CD4陽性細(xì)胞亞群Th17細(xì)胞在真菌性角膜炎中分化，參與其免疫反應(yīng)，Th17細(xì)胞因子（IL-17）在真菌性角膜炎中發(fā)揮重要作用。
[Abstract]:Objective to investigate the expression of TH17 cytokines in fungal keratitis by establishing a mouse model of fungal keratitis. To explore the role of IL-17 and TH17 cells in the immunology of fungal keratitis. Methods BALB/c mice of clean grade were randomly divided into two groups. One group was fungal infection group. The model of Fusarium solanum keratitis was established by corneal surface lens method in the right eye of mice. The other group was treated with PBS instead of fungal solution in the same way as the infected group. The characteristics of keratitis were observed by slit lamp microscope and the pathological changes of cornea were observed by HE staining. The expression of Th17 cytokines (IL-17) and its specific transcription factor ROR 緯 t (ROR 緯 t) was detected by real-time fluorescence quantitative PCR and enzyme linked immunosorbent assay (Elisa). Result 1. All mice models of fungal keratitis were established successfully. The early stage of fungal infection was a diffuse inflammatory reaction of the whole cornea, the infiltration and opacity gradually aggravated, the cornea became gray and haze at 3 days after infection, and the pathological changes were typical at 5 days after infection. The neovascularization around the cornea resulted in central ulcer, and the lesion tended to improve after 7 days. The total course of disease lasted about 12 days. There was statistical difference in clinical score at different time points (P < 0.05). 2. HE staining showed early corneal stroma edema, increased corneal stromal disorder polymorphonuclear cell infiltration and exudation in anterior chamber on the 5th day after infection, neovascularization and erythrocyte in the late corneal stroma. The ulcer is replaced by scar tissue. 3.Real time PCR and ELISA analysis showed that Thl7 cytokines (IL-17) and Th17 cell specific transcription factor (ROR 緯 t) gene mRNA were expressed after fungal infection of cornea. On the 7th day, the expression level was significantly higher than that in the injured control group (P < 0.01), and the highest expression was observed at the 5th day after injury (P < 0.01). The expression of IL-17 factor protein at each time point after infection was significantly higher than that in the injured control group (P < 0. 01). Conclusion Th17 cells, a new type of CD4 positive cell subgroup, differentiate in fungal keratitis and participate in its immune response. Th17 cytokines (IL-17) play an important role in fungal keratitis.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R772.21

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相關(guān)期刊論文 前2條

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