高尿酸血症患者血浆microRNA表达谱分析及其靶基因预测研究

发布时间：2018-08-03 13:11

【摘要】：目的:通过micro RNA基因芯片技术研究无症状高尿酸血症患者与正常对照者血浆中micro RNA的表达差异,并对差异表达的micro RNA进行靶基因预测,探讨micro RNA在高尿酸血症中的作用机制。方法:1.选取4例无症状高尿酸血症患者,4例正常对照者,取其外周血浆用Trizol法提取RNA,使用丹麦ExiqonTM公司的micro RNA芯片检测其血浆中micro RNA表达谱,并筛选出显著差异表达的micro RNA。2.选取1个显著上调的micro RNA及2个显著下调的micro RNA在50例无症状高尿酸血症患者及50例正常对照者中,使用RT-PCR方法进行验证。并对50例高尿酸血症组及50例对照组进行相关临床资料分析。3.通过搜索并汇总miranda数据库、mirbase数据库及targetcsan数据库,对筛选出的差异表达的micro RNA进行靶基因预测,得到特异表达于外周血中的靶基因;再通过GO及Pathway分析,对预测出的靶基因进行相关分子功能分析。结果:1.micro RNA芯片表达结果显示,共有263个micro RNA在高尿酸血症组与对照组间表达有变化,其中差异表达显著的共有11个,上调的有5个,下调的有6个。2.运用RT-PCR技术,与对照组比较,高尿酸血症组中mi R-519d-3p表达上调,差异有统计学意义(P0.01);mi R-374a-5p、mi R-30c-5p表达下调,差异有统计学意义(P0.01),验证结果与micro RNA芯片结果一致。3.通过对显著差异表达的micro RNA的靶基因预测及功能分析,发现这些异常表达的micro RNA主要通过其相关靶基因影响TGF-β信号通路、PI3K-Akt信号通路、细胞内吞作用以及Notch信号通路等来发挥其作用。结论:micro RNA芯片技术是一种有效的高通量分析高尿酸血症micro RNA表达谱的方法,相对于正常对照组,多种micro RNA在高尿酸血症患者外周血浆中出现差异表达,有可能成为高尿酸血症的新的分子生物学标志物并可能参与了高尿酸血症的发病机制,但具体的分子机制还有待于进一步研究。
[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.
【学位授予单位】：青岛大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R589.7

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