dectin1介导的人角上皮细胞对烟曲霉菌的固有免疫反应.docxVIP

优秀毕业论文 精品参考文献资料 Deer Deer i n-1介导的人角膜上皮细胞对烟曲霉菌的固有免疫反应 摘要 目的研究Dectin-I在烟曲霉菌孢子抗原刺激人角膜上皮细胞形成的真菌性角膜炎 模型中的固有免疫反应中的作用。 方法用自制的烟曲霉菌孢子抗原刺激培养的人角膜上皮细胞株，以RT—PCR检测各 组细胞中Dectin-I mRNA的表达；用免疫细胞化学方法及酶联免疫吸附试验(ELISA) 方法检测细胞中Dectin-i蛋白的表达；以ELISA法检测细胞培养上清中NF-K B、 TNF-a、IL—l B、IL-IO和CXCLI的浓度。统计学处理采用SPSS 17．0软件进行单因 素方差分析。 结果 在正常和烟曲霉菌抗原刺激后的人角膜上皮细胞中用RT-PCR均能检测到 Dectin-i mRNA的表达，Dectin-i mRNA表达随刺激时间增长而逐渐增多，且在2h 时达高峰，刺激后各组同空白组相比差距有统计学意义(P=O．000)；用免疫细胞化 学方法检测各组细胞中dectin-I蛋白均有表达；ELISAN得HCECs总蛋白中Dectin-i 正常组有表达，刺激后0．5h蛋白含量下降，1h起升高，2h含量达高峰，随后下降， 刺激后各组同空白组相比差距有统计学意义(P=O．000)；ELISA法检测各组细胞培养 上清中NF-K B刺激后含量增高，2h含量达高峰，刺激后各组同空白组相比差距有统 计学意义(P=O．000)；TNF-Q，CXCLI和IL-I B刺激后含量增高，分别在刺激2h、4h、 1h后达高峰，IL-IO,U激后含量增高缓慢，6h增高达高峰，这四个指标刺激后各组同 空白组相比差距均有统计学意义(P=O．000)。 结论 1、正常的人角膜上皮细胞中存在Dectin-i基因和蛋白水平的表达。 2、用自制的烟曲霉菌孢子抗原刺激正常的人角膜上皮细胞可引起Dectin-i合成量 的变化，提示Dectin-I在该真菌性角膜炎模型中可被激活。 3、NF-KB在该真菌性角膜炎模型中表达增加，同时相关炎症因子TNF-a、IL-I 9、 IL一10和CXCLI的含量变化提示他们也参与了该真菌性角膜炎模型中的免疫反应。 硕士研究生徐强(眼科学) 指导教师 赵桂秋教授 关键词：Dectin-1：固有免疫：人角膜上皮细胞；烟曲霉菌 Activition Activition of Dectin一1 in Aspergillus fumigatus—Induced Innate Immune Responses in Human Cornea Epithelial Cells Abstract Obj ective To investigate expression and role of Dectin．1 and its corresponding factors in human corneal epithelial cells exposed to Aspergillus fumigatus antigens． Methods Stimulated cultured human comeal epithelium cells with Aspergillus fumigatus antigens were collected and expression of Dectin．1 mRNA was done by RT—PCR：expression of Deetin．1 in protein level was done by immunocytochemistry SP method and ELISA；level of expression of relative downstream factors like NF．1(B，TNF．a，IL．1 B，IL．1 0 and CXCL 1 in cell culture media were measured by ELISA．The comparisons among groups were done by SPSS 1 7．0 software． Results Dectin-1 mRNA existed in normal HCECs and increased after fungal stimulation，and its expression upregulated by stimulation time，reaching their peak at 2h，and every stimulated groups were statistically significant compared with blank gr