本文選題：銀屑病 + miRNA�。� 參考：《天津醫(yī)科大學(xué)》2013年碩士論文

【摘要】：目的：利用miRNA芯片技術(shù)分析銀屑病患者與正常對(duì)照者外周血miRNAs表達(dá)譜的差異,同時(shí)對(duì)差異表達(dá)的miRNAs所調(diào)控的靶基因進(jìn)行預(yù)測(cè)和分子功能分析,以探討它們與銀屑病發(fā)病機(jī)制的關(guān)系。 方法：按照入選標(biāo)準(zhǔn),選取進(jìn)行期尋常型銀屑病患者3例,健康自愿者2例,分別進(jìn)入實(shí)驗(yàn)組和正常對(duì)照組。然后采集其血液樣本進(jìn)行總RNA提取,質(zhì)量和純度鑒定；符合Affymetrix miRNA芯片技術(shù)要求后按照miRNA芯片標(biāo)準(zhǔn)實(shí)驗(yàn)流程進(jìn)行熒光標(biāo)記,雜交；然后利用Affymetrix GeneChip Scanner3000激光共聚焦掃描儀對(duì)雜交結(jié)果進(jìn)行圖像掃描,同時(shí)Affymetrix GeneChip Operating Software Versionl.4數(shù)據(jù)處理軟件讀取,處理數(shù)據(jù)；按照q-value(%)≤5,同時(shí)差異倍數(shù)控制在2倍以上的標(biāo)準(zhǔn)篩選差異表達(dá)的miRNAs;利用Human Target Scan5.1數(shù)據(jù)庫(kù)對(duì)差異表達(dá)的miRNAs進(jìn)行靶基因預(yù)測(cè)；運(yùn)用MAS系統(tǒng)的Pathway和GO分類數(shù)據(jù)庫(kù)對(duì)預(yù)測(cè)的靶基因的分子功能進(jìn)行分析。 結(jié)果：實(shí)驗(yàn)組和對(duì)照組相比,差異表達(dá)的miRNAs有5個(gè),其中,hsa-miR-363、 hsa-miR-30e、hsa-miR-192表達(dá)上調(diào),hsa-miR-423-5p和hsa-miR-720表達(dá)下調(diào),它們的長(zhǎng)度均集中在17-23個(gè)核苷酸之間；5個(gè)差異表達(dá)的miRNAs的預(yù)測(cè)靶基因達(dá)到493個(gè)；Pathway分類結(jié)果顯示這些靶基因的功能涉及多種信號(hào)通路的傳導(dǎo),多種腫瘤的發(fā)生、發(fā)展,細(xì)胞凋亡,細(xì)胞因子的相互作用等生物學(xué)過(guò)程；GO分類結(jié)果顯示這些靶基因的分子功能主要是一些酶類、受體、細(xì)胞因子等分子的活性,生物學(xué)過(guò)程主要涉及DNA的復(fù)制、RNA的降解、細(xì)胞凋亡、血管生成、炎性反應(yīng)、免疫應(yīng)答、免疫細(xì)胞的增殖和分化等。 結(jié)論：銀屑病患者外周血中存在差異表達(dá)的miRNAs,其中,hsa-miR-363、 hsa-miR-30e、hsa-miR-192表達(dá)上調(diào),hsa-miR-423-5p和hsa-miR-720表達(dá)下調(diào)；5個(gè)差異表達(dá)的miRNAs的預(yù)測(cè)靶基因達(dá)到493個(gè)；這些差異表達(dá)的miRNAs可能通過(guò)對(duì)其靶基因功能的調(diào)控,而在銀屑病發(fā)病機(jī)制中發(fā)揮重要的調(diào)節(jié)作用。因此,我們的研究可能揭示了銀屑病發(fā)病機(jī)制的一個(gè)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 .

【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R758.63

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