【摘要】：目的:利用miRNA芯片技術(shù)分析銀屑病皮損與正常皮膚組織之間miRNA表達(dá)差異,同時(shí)對(duì)差異表達(dá)的miRNA所調(diào)控的靶基因進(jìn)行預(yù)測和分子功能分析,以及如何參與銀屑病發(fā)生等方面進(jìn)行探討,旨在進(jìn)一步闡述本病發(fā)病機(jī)制的基礎(chǔ)上為臨床的靶向治療提供依據(jù)。方法:本研究選擇尋常型銀屑病進(jìn)行期3例,正常對(duì)照組3例,分別取皮損組織和正常皮膚組織進(jìn)行總RNA提取,質(zhì)量、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)試實(shí)驗(yàn)流程進(jìn)行熒光標(biāo)記,雜交,然后利用Affymetrix GeneChip Scanner 3000激光共聚焦掃描儀對(duì)雜交結(jié)果進(jìn)行圖像掃描,配合 Affymetrix GeneChip Operating Software Version1.4 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)(Fold Change)控制在2倍以上的標(biāo)準(zhǔn)篩選差異表達(dá)的miRNA作為分析數(shù)據(jù)的來源。對(duì)差異表達(dá)的miRNA進(jìn)行靶基因預(yù)測和功能分析分別利用Human Target scan 5.1數(shù)據(jù)庫和MAS系統(tǒng)Pathway和GO分類數(shù)據(jù)庫。結(jié)果:①3例尋常型銀屑病患者皮損組織與3例正常人皮膚組織相比較,差異表達(dá)的miRNA有5條,其中Hsa-miR-1308,表達(dá)上調(diào);Hsa-miR-27a,Hsa-miR-199a-3p,Hsa-miR-199b-3p,Hsa-miR-181a,表達(dá)均下調(diào),5 條差異表達(dá)的miRNA序列長度均集中在18～23個(gè)核苷酸之間。②Human Target scan5.1數(shù)據(jù)庫對(duì)5條差異表達(dá)的miRNA所負(fù)調(diào)控的靶基因進(jìn)行預(yù)測,數(shù)目達(dá)到2168條;利用MAS系統(tǒng)Pathway和GO分類數(shù)據(jù)庫分析2168條靶基因,結(jié)果顯示,這些靶基因主要涉及多種信號(hào)通路的傳導(dǎo),角質(zhì)形成細(xì)胞的增殖與分化,免疫細(xì)胞增殖與調(diào)控等生物學(xué)過程。結(jié)論:銀屑病是一種遺傳基因異常性疾病,轉(zhuǎn)錄水平上的miRNA可靶向負(fù)調(diào)控大量mRNA,而這些被調(diào)控的mRNA之間又存在錯(cuò)綜復(fù)雜的相互影響和作用,共同形成網(wǎng)絡(luò)式的調(diào)節(jié)結(jié)構(gòu)通過多種途徑來參與銀屑病的發(fā)生。
[Abstract]:Objective: to analyze the difference of miRNA expression between psoriatic lesions and normal skin tissues by using miRNA microarray, and to predict and analyze the molecular function of target genes regulated by differentially expressed miRNA. And how to participate in the occurrence of psoriasis were discussed in order to further explain the pathogenesis of psoriasis on the basis of clinical targeted therapy. Methods: in this study, 3 cases of psoriasis vulgaris and 3 cases of normal control group were selected for total RNA extraction, quality and purity identification. According to the technical requirements of Affymetrix miRNA chip, fluorescence labeling and hybridization were carried out according to the experimental procedure of miRNA chip standard, and then the result of hybridization was scanned by Affymetrix GeneChip Scanner 3000 laser confocal scanner. The Affymetrix GeneChip Operating Software Version1.4 data processing software is used to read and process the data, and according to q-value (%) 鈮，

本文編號(hào)：2264524