痛风患者外周血单个核细胞环状RNA的表达谱分析
Circular RNA expression profiles of peripheral blood mononuclear cells by microarray analysis in gout patients
摘要目的:分析环状RNA(circRNA)在痛风患者PBMCs中的表达谱,探讨circRNA在痛风发生发展中可能的作用机制。方法:收集24例急性期痛风患者(AG)、24例间歇期痛风患者(IG)以及24名健康体检者(HC)外周血标本。随机选取3例AG、3例IG、3名HC,采用基因芯片技术筛选其PBMCs中差异表达的circRNA。选择两两对比组间差异倍数较大的6个circRNA,采用实时荧光定量PCR(RT-qPCR)在所有72例研究对象PBMCs中验证了它们的相对表达量。对显著差异表达的circRNA(变化倍数>1.5, P<0.05)进行了基因本体(GO)分析、京都基因与基因组百科全书(KEGG)通路分析以及预测了其与微RNA(miRNA)的相互作用。采用中位数(四分位数间距)描述数据,组间比较采用Mann-Whitney U检验。 结果:(1)芯片数据显示,与HC组相比,AG和IG组分别有116个和41个显著差异表达的circRNA;与IG组相比,AG组有105个显著差异表达的circRNA。(2) RT-qPCR验证的6个circRNA中,5个基因表达趋势与芯片结果一致,其中hsa_circRNA_105034在AG组中表达量相对于IG及HC组差异具有统计学意义[AG组5.17(4.60),IG组1.68(2.39),HC组0.90(0.73),AG组与IG组( Z=-4.413, P<0.01);AG组与HC组( Z=-5.052, P<0.01)]。(3)生物信息学分析:① GO分析发现差异circRNA主要参与DNA的转录调控、细胞的正性调控及蛋白质修饰等;② KEGG通路分析发现它们可能参与了丝裂原活化蛋白激酶(MAPK)信号通路介导的免疫反应;③ circRNA与miRNA的结合位点预测提示痛风中差异表达的circRNA可能通过靶向miRNA-146a、miRNA-302b和miRNA-23a等分子影响其炎症反应。 结论:痛风患者PBMCs中存在差异表达的circRNA,可能与痛风的发生发展密切相关。
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abstractsObjective:To analyze the expression of circular RNA (circRNA) in peripheral blood mononuclear cells (PBMCs) of patients with gout and to explore the possible mechanism of circRNA in the pathogenesis of gout.Methods:Peripheral blood samples of 24 patients with acute gout (AG), 24 patients with intermittent gout (IG) and 24 healthy control subjects (HC) were collected. Three cases of AG, IG, and HC were randomly selected, and the differentially expressed circRNA in PBMCs was screened by human circNA microarrays. The 6 circRNAs with large differences between the two comparison groups were selected, and the relative expression levels of 6 circRNAs in all the collected 72 PBMCs of the study subjects were detected by real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). The significantly differentially ex-pressed circRNA (fold change>1.5, P<0.05) was analyzed by GO analysis, Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis, and its interaction with microRNA (miRNA) was predicted. The median (interquartile range) was used to describe the data, and the Mann-Whitney U test was used for comparison between groups. Results:(1) The microarray analysis results showed that compared with the HC group, the AG group and the IG group had 116 and 41 significantly differently expressed circRNAs, respectively; com-pared with the IG group, the AG group had 105 significantly differently expressed circRNAs. (2) Among the 6 circRNAs verified by PT-qPCR, the expression trends of 5 were consistent with the microarray results. The expression of hsa_circRNA_105034 in the AG group [5.17(4.60)] was statistically significantly different com-pared to the IG [1.68(2.39)] and HC [0.90(0.73)] groups (AG vs IG: Z=-4.413, P<0.01; AG vs HC Z=-5.052, P<0.01). (3) Bioinformatics analysis: ① GO analysis found that differential circRNA swere mainly involved in DNA transcriptional regulation, positive cell regulation and protein modification, etc. ② KEGG pathway analysis revealed that differential circRNA might be involved in the immune response mediated by the mitogen-activated protein kinase signaling pathway. ③ CircRNA might affect its inflammatory response by targeting molecules such as miRNA-146a, miRNA-302b and miRNA-23a. Conclusion:There are differentially expressed circRNAs in PBMCs of patients with gout, which may be closely related to the occurrence and development of gout.
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