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作 者:袁呈晨 耿姗[1] 朱亚梅[2] 彭孝武[1] 周学平[1] 周玲玲[1] YUAN Cheng-Chen GENG Shan ZHU Ya-Mei PENG Xiao-Wu ZHOU Xue-Ping ZHOU Ling-Ling(Nanjing University of Chinese Medicine, Nanjing 210023, China)
机构地区:[1]南京中医药大学,南京210023 [2]南京市中医院,南京210023
出 处:《中国免疫学杂志》2017年第5期715-720,共6页Chinese Journal of Immunology
基 金:国家自然科学基金(81673937);国家基金预研基金(No.14XYY01)资助
摘 要:目的:筛选类风湿关节炎(RA)患者外周血与关节炎大鼠破骨细胞的miRNAs异常表达谱,分析RA的特征性miRNAs及其功能。方法:采用miRNAs芯片,筛选RA患者与正常人外周血、共培养诱生的关节炎大鼠破骨细胞与正常基质细胞的miRNAs表达谱的差异;Real time PCR对芯片结果进行验证;生物信息学方法对关键miRNA进行功能分析。结果:与正常人相比,RA患者外周血中具有差异表达的miRNAs有189个;与单核细胞比较,破骨细胞中具有差异表达的miRNAs有211个,其中miR-15b-5p等10个miRNAs在RA患者外周血及大鼠破骨细胞中均异常表达。Real time PCR验证结果与芯片检测结果具有一致性。生物信息学分析结果显示特征性miRNA的靶基因显著富集在VEGF,MAPK信号通路等信号通路。结论:部分miRNAs在RA患者外周血及关节炎大鼠破骨细胞中的异常表达具有一致性,其可能作为RA的特征性miRNAs,通过调控相关信号通路干预破骨细胞分化等病理过程。Objective:To analyze the different expressions of miRNAs in peripheral blood of rheumatoid arthritis (RA) patients and the osteoclasts in rats,and to verify the key miRNAs in RA.Methods: The miRNAs expressions in monocyte and the co-cultured osteoclasts,peripheral blood of rheumatoid arthritis (RA) patients and normal were detected by miRCURYTM LNA Array.Real time PCR was applied to verify the reliability of miRNA array.The bioinformatics software and database were applied to predict and analyze target genes.Results: miRNA array results showed that 189 miRNAs changed in RA patients compared with the normal;211 miRNAs were changed in osteoclasts group compared with monocytes group.The expressions of ten miRNAs were all abnormal in RA patients and osteoclasts in rats.The results of Real time PCR were consistent with the array.Results of bioinformatics analysis showed that the target gene of miRNA significantly enriched in signaling pathways such as VEGF,MAPK signaling pathways.Conclusion: Some miRNAs had similar abnormal expressions in different species,and these miRNAs may influence the differentiation of the osteoclasts through regulating the related signal passways.
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