糖尿病肾病患者Piwi相互作用RNA差异表达谱分析  

Analysis of differential expression profiles of Piwi-interacting RNA in diabetic nephropathy patients

作　　者：雷寓淇 周思捷 乔颖进 高丹 刘风勋 吕林晓 潘少康 刘东伟 刘章锁 Lei Yuqi;Zhou Sijie;Qiao Yingjin;Gao Dan;Liu Fengxun;Lyu Linxiao;Pan Shaokang;Liu Dongwei;Liu Zhangsuo(Traditional Chinese Medicine Integrated Department of Nephrology,the First Affiliated Hospital of Zhengzhou University,Research Institute of Nephrology,Zhengzhou University,Henan Province Research Center for Kidney Disease,Key Laboratory of Precision Diagnosis and Treatment for Chronic Kidney Disease in Henan Province,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院中西医结合肾病科,郑州大学肾脏病研究所,河南省肾脏病研究中心,河南省慢性肾脏病精准诊疗重点实验室,郑州450052

出　　处：《中华肾脏病杂志》2023年第4期253-262,共10页Chinese Journal of Nephrology

基　　金：国家自然科学基金联合项目(U21A20348);国家自然科学基金青年项目(81800648);河南优秀青年科学基金项目(202300410363)。

摘　　要：目的探究Piwi相互作用RNA(Piwi-interacting RNA,piRNA)与糖尿病肾病(diabetic nephropathy,DN)的相关性。方法使用高通量测序技术检测DN患者肾组织(实验组)及肾肿瘤患者肿瘤旁肾组织(对照组)中piRNA的差异表达谱,通过基因本体(gene ontology,GO)和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)富集分析对差异表达piRNA的生物学功能进行阐述。采用实时荧光定量PCR检测目标piRNA在DN患者血清中的表达水平。采用Spearman相关分析法分析血清目标piRNA与DN患者临床指标之间的相关性。结果高通量测序结果显示,以|log2差异倍数|≥2且P<0.05为筛选条件,与对照组相比,DN组显著差异表达的piRNA共127个,其中99个表达上调,28个表达下调,上调前5位的piRNA分别是piRNA-hsa-161686、piRNA-hsa-349255、piRNA-hsa-355720、piRNA-hsa-151229及piRNA-hsa-154959,下调前5位的piRNA分别是piRNA-hsa-1929960、piRNA-hsa-174194、piRNA-hsa-148658、piRNA-hsa-172594及piRNA-hsa-172421。PCR验证P值较小且表达量较高的3个上调和3个下调基因,结果显示,血清piRNA-hsa-77976的表达在DN患者中显著下调(P=0.028),与测序结果一致,其余基因表达与测序结果不一致或差异无统计学意义。生物信息学分析预测显示显著差异表达的piRNA可能通过Rap1、Ras、PI3K-Akt及轴突导向通路参与DN的调控。相关性分析结果显示,piRNA-hsa-77976表达与血尿素氮(r=-0.584,P=0.028)、血肌酐(r=-0.637,P=0.014)、胱抑素C(r=-0.738,P=0.003)及β2微球蛋白(r=-0.822,P<0.001)均呈负相关,与估算肾小球滤过率(r=0.661,P=0.010)呈正相关。结论piRNA差异表达与DN密切相关,或可作为DN诊断及预后评估的新型生物标志物。Objective To investigate the correlation between Piwi-interacting RNA(piRNA)and diabetic nephropathy(DN).Methods The differential expression profiles of piRNAs in renal tissues of patients with DN(experimental group)and renal tissues adjacent to tumors of patients with renal tumors(control group)were detected by high-throughput sequencing.The biological function of differentially expressed piRNAs was described by gene ontology and Kyoto encyclopedia of genes and genomes enrichment analysis.Real-time fluorescence quantitative PCR was used to detect the serum expression level of target piRNAs in patients with DN.Spearman correlation analysis was used to analyze the correlation between serum target piRNAs and clinical indexes of patients with DN.Results The results of high throughput sequencing showed that there were 127 differentially expressed piRNAs between DN group and control group,with screening condition of|log2(fold changes)|≥2 and P<0.05.Among them,there were 99 up-regulated piRNAs and 28 down-regulated piRNAs.The top 5 up-regulated piRNAs were piRNA-hsa-161686,piRNA-hsa-349255,piRNA-hsa-355720,piRNA-hsa-151229 and piRNA-hsa-154959,respectively.The top 5 down-regulated piRNAs were piRNA-hsa-1929960,piRNA-hsa-174194,piRNA-hsa-148658,piRNA-hsa-172594 and piRNA-hsa-172421,respectively.The PCR verification results of 3 up-regulated genes and 3 down-regulated genes with low P values and high expression levels showed that serum expression level of piRNA-hsa-77976 was significantly down-regulated in patients with DN(P=0.028),which was consistent with that of sequencing,while the expression levels of other genes were inconsistent with the sequencing results or had no statistical significance.Bioinformatics analysis results predicted that significantly differentially expressed piRNAs might participate in the regulation of DN through Rap1,Ras,PI3K-Akt and axon guiding pathways.The results of correlation analysis showed that the expression level of piRNA-hsa-77976 was negatively correlated with blood urea nitrogen(r

关 键 词：糖尿病肾病 RNA/未翻译 RNA/小分子干扰 高通量测序 生物信息学分析 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]