MIP-2、MMP-2与TIMP-2在兔棘阿米巴性角膜炎的表达

发布时间：2018-04-18 17:56

本文选题：棘阿米巴性角膜炎 + 巨噬细胞炎性蛋白-2　；参考：《福建医科大学》2011年硕士论文

【摘要】：【摘要】目的:建立有效的棘阿米巴性角膜炎的动物模型,观察棘阿米巴性角膜炎模型眼的发展过程及临床表现,于病程的不同阶段行病理组织学检查,并检测巨噬细胞炎性蛋白-2(MIP-2)与基质金属蛋白酶-2(MMP-2)及其抑制剂(TIMP-2)的表达,探讨其在棘阿米巴角膜炎发病机制中的作用。方法:新西兰大白兔35只,随机分为7组,每组5只,其中1组为正常对照组,6组为实验组,右眼为实验眼,实验组采用角膜表面镜片术辅助上皮刮除术建立兔棘阿米巴角膜炎模型,感染术后缝合兔睑裂,24h后打开眼睑,每天观察角膜感染情况,于不同时间点利用免疫组织化学染色法检测MIP-2,MMP-2、TIMP-2在角膜的表达,利用图像分析系统进行半定量分析。结果:(1)经角膜刮片及组织病理切片检查证实实验组30只兔子右眼均感染棘阿米巴性角膜炎。(2)HE染色观察角膜炎症反应发生在感染早期,随着病程的进展,炎症细胞逐渐增多,在感染后第5d达到高峰,7d后逐渐呈下降趋势,角膜组织开始呈现修复状态。(3)实验组MIP-2主要表达于受损的角膜上皮层和基质层,正常对照组角膜组织未见其表达。在实验组中,角膜感染棘阿米巴后1天即检测到MIP-2活性,5天活性到达高峰,之后逐渐下降。(4)实验组MMP-2,TIMP-2阳性表达细胞的平均吸光值(A)与正常对照组相比差异有统计学意义(P0.05),即MMP-2,TIMP-2在棘阿米巴性角膜炎中的表达高于正常角膜,且主要位于角膜基质层。在实验组中,MMP-2,TIMP-2活性均在第5天开始升高,第9天达到高峰,半定量分析中TIMP-2表达弱于MMP-2。结论:(1)利用角膜表面镜片术辅助上皮刮除术可建立有效、可靠的棘阿米巴性角膜炎动物模型,且该建模方式符合该疾病的自然病程;(2)棘阿米巴角膜炎的早期MIP-2的表达明显增加,且表达量与角膜炎的炎症反应呈正相关,这表明MIP-2作为重要的趋化因子参与了棘阿米巴角膜炎的免疫防御反应;(3)MMP-2,TIMP-2主要参与棘阿米巴角膜炎后期的自身修复反应,MMP-2相对于TIMP-2过量表达,导致角膜细胞外基质的降解,,MMP-2/TIMP-2系统的失衡导致角膜基质层的损伤。
[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.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R772.21

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