本文選題：煙曲霉菌 + 角膜上皮細(xì)胞��； 參考：《青島大學(xué)》2012年碩士論文

【摘要】：目的觀察白細(xì)胞介素6(interleukin-6,-6)和白細(xì)胞介素10(interleukin-0,IL-10)在大鼠煙曲霉菌性角膜炎固有免疫階段角膜上皮組織中的表達(dá)并分析其表達(dá)量變化的原因。 方法健康Wi star大鼠72只,雌雄不限,在刮除中央?yún)^(qū)角膜上皮的大鼠角膜上涂以煙曲霉菌標(biāo)準(zhǔn)菌落并覆蓋自制角膜接觸鏡,建立大鼠煙曲霉菌性角膜炎感染模型。大鼠隨機(jī)分為2部分,用于免疫組織化學(xué)實(shí)驗(yàn)的A1、B1、C1組(每組各6只大鼠)和用于RT-PCR實(shí)驗(yàn)的A2、B2、C2(A2組6只、B2、C2組24只)：A1、A2組為空白對照組,不予任何處理；B1、B2組為損傷對照組,建立模型,僅刮除中央?yún)^(qū)角膜上皮后覆蓋角膜接觸鏡,不涂抹真菌；C1、C2組為真菌模型組,建立真菌模型。成功建模后4h打開眼瞼,并于造模后第4h、8h、16h、24h處死大鼠,取角膜上皮組織。HE染色觀察角膜組織病理變化并檢測真菌生長,運(yùn)用免疫組織化學(xué)和RT-PCR技術(shù)檢測各組不同時間點(diǎn)IL-6與IL-10的表達(dá)情況。 結(jié)果1.組織病理學(xué)觀察見真菌性角膜炎病灶內(nèi)有炎癥細(xì)胞浸潤,HE染色檢查在角膜基質(zhì)層可見菌絲及孢子。2.IL-6在空白對照眼角膜基本不表達(dá)或極微量表達(dá)；真菌實(shí)驗(yàn)組在接種真菌后4h開始出現(xiàn)IL-6的表達(dá),并逐漸增高,在第24h表達(dá)量升高顯著,各時間點(diǎn)比較差異具有顯著性(P0.01)。3.IL-10在空白對照組角膜基本不表達(dá)或極微量表達(dá)；真菌實(shí)驗(yàn)組在真菌刺激后4h開始出現(xiàn)IL-10的表達(dá),并逐漸增高,在24h表達(dá)量升高顯著,各時間點(diǎn)比較差異有顯著性((P0.01)。 結(jié)論1.10%氫氧化鉀涂片及組織刮取物行菌落培養(yǎng)證實(shí)大鼠真菌性角膜炎動物模型建立成功。2.IL-6在煙曲霉菌性角膜炎角膜上皮組織中早期即開始表達(dá),表達(dá)量逐漸增高,其表達(dá)量的增高與角膜炎癥程度呈正相關(guān),在大鼠煙曲霉菌性角膜炎病程中是一個敏感的炎癥因子,能反映局部的炎癥反應(yīng)程度。3.IL-10表達(dá)于真菌性角膜炎角膜上皮組織中,煙曲霉菌菌絲刺激后表達(dá)升高,24h升高顯著,提示IL-10可能參與了煙曲霉菌性角膜炎的發(fā)生與發(fā)展,作為一種保護(hù)性細(xì)胞因子通過抗炎抗免疫反應(yīng)也參與了角膜組織修復(fù)和抗損傷過程。
[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.
【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】：2012
【分類號】：R772.21

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 杜兆東;梁濤;趙桂秋;劉建偉;張麗娜;張振華;;真菌性角膜炎病原菌的流行病學(xué)調(diào)查[J];青島大學(xué)醫(yī)學(xué)院學(xué)報;2007年02期

2 張彩;田志剛;;天然免疫識別機(jī)制及天然免疫受體的相互調(diào)節(jié)[J];生物化學(xué)與生物物理進(jìn)展;2008年02期

相關(guān)博士學(xué)位論文 前3條

1 蔡學(xué)敏;甘露聚糖結(jié)合凝集素與相關(guān)絲氨酸蛋白酶相互作用的初步研究[D];第一軍醫(yī)大學(xué);2007年

2 趙杰;Toll樣受體信號通路在角膜煙曲霉菌感染中的作用[D];山東大學(xué);2008年

3 王明永;甘露聚糖結(jié)合凝集素調(diào)節(jié)獲得性免疫應(yīng)答的初步研究[D];南方醫(yī)科大學(xué);2008年

相關(guān)碩士學(xué)位論文 前1條

1 李娜;IL-10在大鼠真菌性角膜炎中的表達(dá)[D];青島大學(xué);2010年

，

本文編號：1824730