银屑病患者外周血microRNA差异表达的研究

发布时间：2018-05-14 08:17

本文选题：银屑病 + miRNA　；参考：《天津医科大学》2013年硕士论文

【摘要】：目的：利用miRNA芯片技术分析银屑病患者与正常对照者外周血miRNAs表达谱的差异,同时对差异表达的miRNAs所调控的靶基因进行预测和分子功能分析,以探讨它们与银屑病发病机制的关系。方法：按照入选标准,选取进行期寻常型银屑病患者3例,健康自愿者2例,分别进入实验组和正常对照组。然后采集其血液样本进行总RNA提取,质量和纯度鉴定；符合Affymetrix miRNA芯片技术要求后按照miRNA芯片标准实验流程进行荧光标记,杂交；然后利用Affymetrix GeneChip Scanner3000激光共聚焦扫描仪对杂交结果进行图像扫描,同时Affymetrix GeneChip Operating Software Versionl.4数据处理软件读取,处理数据；按照q-value(%)≤5,同时差异倍数控制在2倍以上的标准筛选差异表达的miRNAs;利用Human Target Scan5.1数据库对差异表达的miRNAs进行靶基因预测；运用MAS系统的Pathway和GO分类数据库对预测的靶基因的分子功能进行分析。结果：实验组和对照组相比,差异表达的miRNAs有5个,其中,hsa-miR-363、 hsa-miR-30e、hsa-miR-192表达上调,hsa-miR-423-5p和hsa-miR-720表达下调,它们的长度均集中在17-23个核苷酸之间；5个差异表达的miRNAs的预测靶基因达到493个；Pathway分类结果显示这些靶基因的功能涉及多种信号通路的传导,多种肿瘤的发生、发展,细胞凋亡,细胞因子的相互作用等生物学过程；GO分类结果显示这些靶基因的分子功能主要是一些酶类、受体、细胞因子等分子的活性,生物学过程主要涉及DNA的复制、RNA的降解、细胞凋亡、血管生成、炎性反应、免疫应答、免疫细胞的增殖和分化等。结论：银屑病患者外周血中存在差异表达的miRNAs,其中,hsa-miR-363、 hsa-miR-30e、hsa-miR-192表达上调,hsa-miR-423-5p和hsa-miR-720表达下调；5个差异表达的miRNAs的预测靶基因达到493个；这些差异表达的miRNAs可能通过对其靶基因功能的调控,而在银屑病发病机制中发挥重要的调节作用。因此,我们的研究可能揭示了银屑病发病机制的一个新的分子水平。
[Abstract]:Objective : To analyze the difference of expression profiles of peripheral blood from patients with psoriasis and normal controls by miRNA chip technique , and to predict and analyze the target genes regulated by differentially expressed miRNA in order to explore their relationship with pathogenesis of psoriasis .

Methods : According to the inclusion criteria , 3 patients with psoriasis vulgaris and 2 healthy volunteers were selected to enter the experimental group and the normal control group respectively . Then the blood samples were collected for total RNA extraction , quality and purity identification .
fluorescence labeling and hybridization are carried out according to the miRNA chip standard experimental flow after conforming to the technical requirements of the Affyary miRNA chip ;
The hybridization results were then subjected to image scanning using the AffyTM GeneChip Scanner3000 laser confocal scanner , and the data was read and processed by AffyTM GeneChip Operating Software Version 1.4 data processing software ;
according to q - value ( % ) & lt ; = 5 , at the same time , the multiple controls over 2 times of the standard screening difference expression are controlled , and target gene prediction is carried out on the differentially expressed miRNA by using the Human Target Scan5.1 database ;
The molecular function of the predicted target gene was analyzed by using the Pathway and GO classification database of MAS system .

Results : The expression of hsa - miR - 363 , hsa - miR - 30 , hsa - miR - 192 , hsa - miR - 423 - 5p and hsa - miR - 720 were down - regulated in the experimental group and the control group .
The predicted target gene of 5 differentially expressed miRNA was 493 ;
Pathway classification results show that the functions of these target genes involve the conduction of various signal pathways , the occurrence , development , cell apoptosis , the interaction of cytokines and other biological processes .
GO classification results show that the molecular functions of these target genes are mainly the activity of some enzymes , receptors , cytokines , etc . The biological process mainly involves DNA replication , RNA degradation , apoptosis , angiogenesis , inflammatory response , immune response , proliferation and differentiation of immune cells , etc .

Conclusion : The expression of hsa - miR - 363 , hsa - miR - 30 , hsa - miR - 192 is up - regulated and hsa - miR - 423 - 5p and hsa - miR - 720 are downregulated in patients with psoriasis .
The predicted target gene of 5 differentially expressed miRNA was 493 ;
The expression of these differences may play an important role in the pathogenesis of psoriasis by regulating its target gene function . Therefore , our study may reveal a new molecular level in the pathogenesis of psoriasis .

【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R758.63

【参考文献】