活化巨噬细胞防治真菌性角膜炎的实验研究

发布时间：2018-05-08 22:09

本文选题：真菌性角膜炎 + 乳胶微粒　；参考：《福建医科大学》2011年硕士论文

【摘要】：目的通过预先Balb/c小鼠中央角膜基质层内注射乳胶微粒活化巨噬细胞,观察真菌性角膜炎的发生发展及其细胞因子在小鼠角膜中的表达水平,探讨机体免疫对小鼠真菌性角膜炎发生发展的影响。方法50只Balb/c小鼠随机分成两组,每组25只。其中1组为实验组,小鼠右眼感染真菌前7天于角膜浅基质层内注入乳胶微粒,然后应用角膜表层镜法建立Balb/c小鼠茄病镰刀菌性角膜炎模型;另1组为对照组,小鼠右眼感染真菌前7天于角膜浅基质层内注入PBS。在感染后第1、3、5、7天,裂隙灯显微镜观察角膜炎特点;HE染色观察角膜病理变化;Real Time PCR和ELISA检测角膜中Thl细胞因子(IL-12)及Th2细胞因子(IL-4)基因mRNA和蛋白的表达。结果1.实验组,早期角膜浸润混浊且进行性加重,主要表现在角膜中央;第5天后新生血管大量生长、角膜穿孔率增高。对照组,早期角膜浸润混浊主要表现为弥漫性病变,第5天后新生血管生长、角膜穿孔,穿孔率较低; 2.HE染色及免疫组化实验组可见早期角膜中央炎症细胞浸润,第5天后大量炎症细胞浸润,角膜基质中纤维细胞减少,角膜后弹力层膨出。对照组可见早期在角膜缘、角膜基质及前房中有大量的炎症细胞浸润,第5天后炎症细胞减少,角膜基质中纤维细胞逐渐增多;免疫组化示实验组较对照组角膜中更多量巨噬细胞浸润。 3.Real time-PCR与ELISA检测结果表明,Thl细胞因子(IL-12)和Th2细胞因子(IL-4)基因mRNA及蛋白在角膜接种菌液后实验组与对照组均出现表达,且IL-12的表达显著强于IL-4(P0.05),但是实验组中Thl细胞因子(IL-12)的表达显著强于对照组。结论在真菌性角膜炎中,Thl/Th2型免疫应答共同参与调节机体的抗真菌免疫,但以Th1型应答为主;角膜感染真菌后第5天机体的免疫力达最强;巨噬细胞在真菌性角膜炎中起重要作用,活化巨噬细胞可能上调免疫反应而加剧真菌性角膜炎炎症反应。
[Abstract]:Objective to observe the occurrence and development of fungal keratitis and the expression of cytokines in the cornea of Balb/c mice by injecting latex particles into the central stromal layer to activate macrophages. To investigate the effect of immune system on the occurrence and development of fungal keratitis in mice. Methods 50 Balb/c mice were randomly divided into two groups with 25 mice in each group. The first group was the experimental group, 7 days before the fungal infection in the right eye, latex particles were injected into the superficial stromal layer of the cornea, then the Balb/c mouse model of Fusarium solani keratitis was established by corneal surface microscopy, the other group was used as the control group. PBSs were injected into the superficial stromal layer of cornea 7 days before fungal infection in the right eye of mice. On the 7th day after infection, keratitis was observed by slit lamp microscope. The expression of Thl cytokine IL-12) and Th2 cytokine IL-4 gene mRNA and protein were detected by real Time PCR and ELISA. Result 1. In the experimental group, the early corneal infiltration was turbid and gradually aggravated, mainly in the central cornea, and the rate of corneal perforation was increased after 5 days of neovascularization. In the control group, early corneal infiltration and opacity mainly showed diffuse lesions, and the rate of corneal perforation was low after 5 days of neovascularization. 2.HE staining and immunohistochemical staining showed early infiltration of central corneal inflammatory cells. After 5 days, a large number of inflammatory cells were infiltrated, fibroblasts in corneal stroma decreased, and the posterior elastic layer of cornea expanded. In the control group, a large number of inflammatory cells were infiltrated in the limbus cornea, corneal stroma and anterior chamber at the early stage. After 5 days, the inflammatory cells decreased and the fibroblasts in the corneal stroma increased gradually. Immunohistochemistry showed that more macrophages infiltrated the cornea in the experimental group than in the control group. The results of 3.Real time-PCR and ELISA showed that the mRNA and protein of THL cytokine IL-12 and Th2 cytokine IL-4 were expressed in both experimental group and control group after corneal inoculation. The expression of IL-12 was significantly higher than that of IL-4 P 0.05, but the expression of Thl cytokine IL-12 was significantly higher in the experimental group than that in the control group. Conclusion Thl / Th2 type immune response in fungal keratitis is involved in the regulation of antifungal immunity, but mainly in Th1 type, and the immunity of the body is strongest on the 5th day after fungal infection. Macrophages play an important role in fungal keratitis. Activated macrophages may up-regulate the immune response and aggravate the inflammatory response of fungal keratitis.
【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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