【摘要】：目的:通過(guò)micro RNA基因芯片技術(shù)研究無(wú)癥狀高尿酸血癥患者與正常對(duì)照者血漿中micro RNA的表達(dá)差異,并對(duì)差異表達(dá)的micro RNA進(jìn)行靶基因預(yù)測(cè),探討micro RNA在高尿酸血癥中的作用機(jī)制。方法:1.選取4例無(wú)癥狀高尿酸血癥患者,4例正常對(duì)照者,取其外周血漿用Trizol法提取RNA,使用丹麥ExiqonTM公司的micro RNA芯片檢測(cè)其血漿中micro RNA表達(dá)譜,并篩選出顯著差異表達(dá)的micro RNA。2.選取1個(gè)顯著上調(diào)的micro RNA及2個(gè)顯著下調(diào)的micro RNA在50例無(wú)癥狀高尿酸血癥患者及50例正常對(duì)照者中,使用RT-PCR方法進(jìn)行驗(yàn)證。并對(duì)50例高尿酸血癥組及50例對(duì)照組進(jìn)行相關(guān)臨床資料分析。3.通過(guò)搜索并匯總miranda數(shù)據(jù)庫(kù)、mirbase數(shù)據(jù)庫(kù)及targetcsan數(shù)據(jù)庫(kù),對(duì)篩選出的差異表達(dá)的micro RNA進(jìn)行靶基因預(yù)測(cè),得到特異表達(dá)于外周血中的靶基因;再通過(guò)GO及Pathway分析,對(duì)預(yù)測(cè)出的靶基因進(jìn)行相關(guān)分子功能分析。結(jié)果:1.micro RNA芯片表達(dá)結(jié)果顯示,共有263個(gè)micro RNA在高尿酸血癥組與對(duì)照組間表達(dá)有變化,其中差異表達(dá)顯著的共有11個(gè),上調(diào)的有5個(gè),下調(diào)的有6個(gè)。2.運(yùn)用RT-PCR技術(shù),與對(duì)照組比較,高尿酸血癥組中mi R-519d-3p表達(dá)上調(diào),差異有統(tǒng)計(jì)學(xué)意義(P0.01);mi R-374a-5p、mi R-30c-5p表達(dá)下調(diào),差異有統(tǒng)計(jì)學(xué)意義(P0.01),驗(yàn)證結(jié)果與micro RNA芯片結(jié)果一致。3.通過(guò)對(duì)顯著差異表達(dá)的micro RNA的靶基因預(yù)測(cè)及功能分析,發(fā)現(xiàn)這些異常表達(dá)的micro RNA主要通過(guò)其相關(guān)靶基因影響TGF-β信號(hào)通路、PI3K-Akt信號(hào)通路、細(xì)胞內(nèi)吞作用以及Notch信號(hào)通路等來(lái)發(fā)揮其作用。結(jié)論:micro RNA芯片技術(shù)是一種有效的高通量分析高尿酸血癥micro RNA表達(dá)譜的方法,相對(duì)于正常對(duì)照組,多種micro RNA在高尿酸血癥患者外周血漿中出現(xiàn)差異表達(dá),有可能成為高尿酸血癥的新的分子生物學(xué)標(biāo)志物并可能參與了高尿酸血癥的發(fā)病機(jī)制,但具體的分子機(jī)制還有待于進(jìn)一步研究。
[Abstract]:Aim: to study the difference of micro RNA expression in plasma between asymptomatic hyperuricemia patients and normal controls by micro RNA gene chip technique, and to predict the target gene expression of micro RNA in order to explore the mechanism of micro RNA in hyperuricemia. Method 1: 1. Four patients with asymptomatic hyperuricemia and 4 normal controls were selected and their peripheral blood plasma was extracted by Trizol method. The expression profiles of micro RNA in plasma were detected by micro RNA microarray of ExiqonTM Company in Denmark, and the differentially expressed micro RNA.2was screened out. One significantly up-regulated micro RNA and two significantly down-regulated micro RNA were tested by RT-PCR in 50 asymptomatic hyperuricemia patients and 50 normal controls. The clinical data of 50 cases of hyperuricemia and 50 cases of control group were analyzed. By searching and summarizing miranda database and targetcsan database, the target gene of differentially expressed micro RNA was predicted, and then the target gene expressed in peripheral blood was obtained, and then analyzed by go and Pathway. The predicted target gene was analyzed by molecular function analysis. Results: 1. The results of RNA microarray showed that there were changes in the expression of 263 micro RNA between hyperuricemia group and control group, of which 11 were significantly different, 5 were up-regulated, and 6. 2 were down-regulated. Using RT-PCR technique, the expression of mi R-519d-3p was up-regulated in hyperuricemia group (P0.01), and the difference was statistically significant (P0.01). The difference was statistically significant (P0.01). The result was consistent with that of micro RNA chip. Through the prediction and functional analysis of the target genes of significantly differentially expressed micro RNA, it was found that the abnormal expression of micro RNA affected the PI3K-Akt signaling pathway mainly through its related target genes. Endocytosis and Notch signaling pathway play their roles. Conclusion the micro RNA microarray technique is an effective method for high-throughput analysis of micro RNA expression profiles in hyperuricemia patients. Compared with the control group, the differential expression of micro RNA was found in peripheral plasma of hyperuricemia patients. It may be a new molecular biomarker of hyperuricemia and may be involved in the pathogenesis of hyperuricemia, but the specific molecular mechanism remains to be further studied.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：R589.7

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