树鼩细菌性角膜炎感染模型的建立及IL-17在模型中的表达

目的:建立树鼩铜绿假单胞菌和金黄色葡萄球菌性角膜炎感染模型,并观察IL-17在模型中的表达,初步探讨IL-17在树鼩细菌性角膜炎中的作用。方法:采用接触镜辅助的角膜划痕法建立树鼩细菌性角膜炎模型。在进行接种后第1、4、7、14d使用前段照相、活体共聚焦显微镜对模型感染症状进行评估,制作病理切片观察角膜组织病理学改变。在相应时间点进行取材,实时荧光定量PCR检测树鼩角膜组织中IL-17 mRNA的表达,ELISA检测IL-17蛋白表达水平。结果:树鼩铜绿假单胞菌和金黄色葡萄球菌性角膜炎模型造模成功率为96%、100%,树鼩细菌性角膜炎临床表现、炎症细胞浸润情况和组织病理改变规律相吻合。IL-17基因和蛋白的表达情况在树鼩角膜中与角膜炎症的严重情况一致。结论:采用接触镜辅助的角膜划痕法可成功建立更接近人细菌性角膜炎自然感染病程的树鼩铜绿假单胞菌和金黄色葡萄球菌性角膜炎动物模型。IL-17参与树鼩细菌性角膜炎发生发展的过程。

Establishment of experimental bacterial keratitis in a tree shrew infection model and expression of interleukin-17 in the model

AIM:To establish the model of pseudomonas aeruginosa and staphylococcus aureus keratitis infection in tree shrews. To determine the expression of IL-17 in the bacterial infection process of tree shrews cornea, and the mechanism of IL-17 in bacterial keratitis of tree shrews is discussed.

METHODS: The tree shrew bacterial keratitis models were established by the contact lens-assisted corneal scratching method. After establishing models successfully, the infection symptoms of the model were evaluated by using anterior segment photography and in vivo confocal microscopy on 1, 4, 7 and 14d after performing inoculation, and pathological sections were made to observe histopathological changes in the cornea. Samples were collected at the corresponding time points above, and the expression of IL-17 mRNA in the corneal tissues of tree shrews was detected by real-time quantitative PCR, and the expression of IL-17 protein was detected by ELISA.

RESULTS:The success rate of modeling the tree shrew pseudomonas aeruginosa and staphylococcus aureus keratitis models was 96% and 100%.The clinical manifestations and inflammatory cell infiltration of the tree shrew keratitis was consistent with the changing rules of the cornea in histopathological. IL-17 gene and protein expression profiles in tree shrew corneas were consistent with the severity of corneal inflammation basically.

CONCLUSION:The use of contact lens-assisted corneal scratching method can successfully establish animal models of pseudomonas aeruginosa and staphylococcus aureus keratitis in tree shrews that more closely resemble the natural course of human bacterial keratitis infection. IL-17 participated in the occurrence and development of bacterial keratitis in tree shrews.