本文選題：棘阿米巴性角膜炎 + 巨噬細(xì)胞炎性蛋白-2　； 參考：《福建醫(yī)科大學(xué)》2011年碩士論文

【摘要】：【摘要】目的:建立有效的棘阿米巴性角膜炎的動物模型,觀察棘阿米巴性角膜炎模型眼的發(fā)展過程及臨床表現(xiàn),于病程的不同階段行病理組織學(xué)檢查,并檢測巨噬細(xì)胞炎性蛋白-2(MIP-2)與基質(zhì)金屬蛋白酶-2(MMP-2)及其抑制劑(TIMP-2)的表達(dá),探討其在棘阿米巴角膜炎發(fā)病機(jī)制中的作用。 方法:新西蘭大白兔35只,隨機(jī)分為7組,每組5只,其中1組為正常對照組,6組為實驗組,右眼為實驗眼,實驗組采用角膜表面鏡片術(shù)輔助上皮刮除術(shù)建立兔棘阿米巴角膜炎模型,感染術(shù)后縫合兔瞼裂,24h后打開眼瞼,每天觀察角膜感染情況,于不同時間點利用免疫組織化學(xué)染色法檢測MIP-2,MMP-2、TIMP-2在角膜的表達(dá),利用圖像分析系統(tǒng)進(jìn)行半定量分析。 結(jié)果:(1)經(jīng)角膜刮片及組織病理切片檢查證實實驗組30只兔子右眼均感染棘阿米巴性角膜炎。(2)HE染色觀察角膜炎癥反應(yīng)發(fā)生在感染早期,隨著病程的進(jìn)展,炎癥細(xì)胞逐漸增多,在感染后第5d達(dá)到高峰,7d后逐漸呈下降趨勢,角膜組織開始呈現(xiàn)修復(fù)狀態(tài)。(3)實驗組MIP-2主要表達(dá)于受損的角膜上皮層和基質(zhì)層,正常對照組角膜組織未見其表達(dá)。在實驗組中,角膜感染棘阿米巴后1天即檢測到MIP-2活性,5天活性到達(dá)高峰,之后逐漸下降。(4)實驗組MMP-2,TIMP-2陽性表達(dá)細(xì)胞的平均吸光值(A)與正常對照組相比差異有統(tǒng)計學(xué)意義(P0.05),即MMP-2,TIMP-2在棘阿米巴性角膜炎中的表達(dá)高于正常角膜,且主要位于角膜基質(zhì)層。在實驗組中,MMP-2,TIMP-2活性均在第5天開始升高,第9天達(dá)到高峰,半定量分析中TIMP-2表達(dá)弱于MMP-2。 結(jié)論:(1)利用角膜表面鏡片術(shù)輔助上皮刮除術(shù)可建立有效、可靠的棘阿米巴性角膜炎動物模型,且該建模方式符合該疾病的自然病程;(2)棘阿米巴角膜炎的早期MIP-2的表達(dá)明顯增加,且表達(dá)量與角膜炎的炎癥反應(yīng)呈正相關(guān),這表明MIP-2作為重要的趨化因子參與了棘阿米巴角膜炎的免疫防御反應(yīng);(3)MMP-2,TIMP-2主要參與棘阿米巴角膜炎后期的自身修復(fù)反應(yīng),MMP-2相對于TIMP-2過量表達(dá),導(dǎo)致角膜細(xì)胞外基質(zhì)的降解,,MMP-2/TIMP-2系統(tǒng)的失衡導(dǎo)致角膜基質(zhì)層的損傷。
[Abstract]:Objective: to establish an effective animal model of Acanthamoeba keratitis, observe the development process and clinical manifestation of Acanthamoeba keratitis model, and make histopathological examination at different stages of disease course.The expression of macrophage inflammatory protein-2 (MIP-2) and matrix metalloproteinase-2 (MMP-2) and its inhibitor, TIMP-2) were detected to explore its role in the pathogenesis of Acanthamoeba keratitis.Methods: Thirty-five New Zealand white rabbits were randomly divided into 7 groups with 5 rabbits in each group.In the experimental group, the rabbit model of Acanthoamoeba keratitis was established by corneal surface lens assisted with epithelial curettage. The eyelid was opened 24 hours after suture of rabbit eyelid fissure after infection, and the corneal infection was observed every day.The expression of MMP-2TIMP-2 in cornea was detected by immunohistochemical staining at different time points, and semi-quantitative analysis was carried out by image analysis system.Results the corneal scrapes and histopathological sections of 30 rabbits in the experimental group were confirmed to be infected with Acanthamoeba keratitis. The corneal inflammatory reaction was observed at the early stage of infection. With the progression of the disease, the inflammatory cells increased gradually.The expression of MIP-2 in the experimental group was mainly in the damaged corneal epithelial layer and stromal layer, but not in the normal control group.In the experimental group, the activity of MIP-2 reached its peak 5 days after infection with Acanthamoeba.The average absorptivity of MMP-2TIMP-2 positive cells in the experimental group was significantly higher than that in the control group (P 0.05), that is, the expression of MMP-2TIMP-2 in Acanthamoeba keratitis was higher than that in the normal cornea, and was mainly located in the corneal stroma.In the experimental group, the activity of MMP-2 and TIMP-2 began to increase on the 5th day and reached the peak on the 9th day. The expression of TIMP-2 in semi-quantitative analysis was weaker than that in MMP-2.Conclusion (1) the effective and reliable animal model of Acanthamoeba keratitis can be established by using corneal surface lens assisted epithelial curettage, and the expression of MIP-2 in the early stage of Acanthamoeba keratitis is significantly increased in accordance with the natural course of the disease.The expression level was positively correlated with the inflammatory response of keratitis.It is suggested that MIP-2, as an important chemokine, is involved in the immune defense response of Acanthamoeba keratitis and the expression of MMP-2 relative to TIMP-2 is mainly involved in the autorepair response of Acanthamoeba keratitis in the late stage of Acanthamoeba keratitis.The imbalance of MMP-2 / TIMP-2 system leads to corneal stromal layer injury.
【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】：2011
【分類號】：R772.21

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