囊液外泌体microRNAs在常染色体显性遗传多囊肾病及单纯性肾囊肿鉴别诊断中的价值  

Value of cystic fluid exosomes-derived microRNAs in differential diagnosis of autosomal dominant polycystic kidney disease and simple renal cystic patients

作　　者：鲍妍 刘向东[1] 王树龙[1] 王云[1] 李瑾[1] BAO Yan;LIU Xiang-dong;WANG Shu-long;WANG Yun;LI Jin(Hospital of Nephrology,the First Affiliated Hospital of Xinxiang Medical University,Weihui,Henan 453100,China)

机构地区：[1]新乡医学院第一附属医院肾脏病医院,河南卫辉453100

出　　处：《医药论坛杂志》2024年第4期337-343,共7页Journal of Medical Forum

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20220591);新乡市科技攻关计划项目(GG2021028);新乡医学院第一附属医院青年培育基金项目(QN-2019-B16)。

摘　　要：目的探索囊液外泌体microRNAs在鉴别常染色体显性遗传多囊肾病(autosomal dominant polycystic kid-ney disease,ADPKD)及单纯性肾囊肿(simple renal cystic,SRC)中的价值。方法选取2019年1月至2021年1月就诊于新乡医学院第一附属医院的初诊ADPKD患者20例作为试验组(ADPKD组),选取同时期年龄性别匹配的SRC患者20例为对照组(SRC组),排除相关禁忌症后完成肾囊肿穿刺抽吸术,分离提取囊液外泌体,每组随机选择6例进行microRNA高通量测序比较组间差异,并采用实时荧光定量PCR验证测序结果。其余每组14例患者作为验证队列,通过ROC曲线来评价差异表达的microRNAs在ADPKD与SRC鉴别诊断中的价值。结果两组研究对象性别、年龄、肌酐、血红蛋白水平差异无统计学意义(P>0.05)。经高通量测序发现,共有18种microRNA在两组间差异表达。与SRC组相比,ADPKD组有9种microRNA表达上调,9种microRNA表达下调。生物信息学分析提示显著上调的microRNA主要涉及近端小管碳酸氢盐重吸收通路、谷氨酰胺代谢通路、干细胞分化通路等通路,显著下调的microRNA主要涉及MAPK信号通路、细胞黏附信号通路等通路。qRT-PCR验证miR-1291、miR-6852-5p的组间表达变化与测序结果一致(P<0.05)。受试者操作特征(ROC)曲线结果显示,囊液外泌体miR-1291(AUC=0.73)、miR-6852-5p(AUC=0.84)具有鉴别ADPKD与SRC的潜力,两者联合检测可提高效能(AUC=0.91)(P<0.05)。结论ADPKD患者囊液外泌体microRNA表达谱与SRC人群相比发生变化,其中差异表达的囊液外泌体miR-1291及miR-6852-5p有望成为鉴别ADPKD及SRC的标记物。Objective To explore the value of cystic fluid exosomes-derived microRNAs in identifying of ADPKD and SRC patients.Methods Twenty newly diagnosed ADPKD patients admitted to the First Affiliated Hospital of Xinxiang Medical University from January 2019 to January 2021 were selected as the experimental group(ADPKD group).20 pa-tients with age-gender matched simple renal cystic(SRC)during the same period were selected as the control group(SRC group).Renal cyst aspiration was performed after exclusion of relevant contraindications.Cyst fluid exosomes were isolated and extracted.Six patients in each group were randomly selected for high-throughput microRNA sequencing to compare the differences between the groups,and real-time fluorescence quantitative PCR(qRT-PCR)was used to verify the sequencing results.The remaining 14 patients in each group were used as a validation cohort to evaluate the value of differentially expressed microRNAs in the differential diagnosis of ADPKD and SRC by receiver-operating characteristic curve(ROC).Results There was no significant difference in gender,age,creatinine and hemoglobin levels between the two groups(P>0.05).The results of high throughput sequencing showed that there were 18 differentially expressed microRNA between ADPKD group and SRC group.Compared with the SRC group,9 microRNA were up-regulated and 9 microRNA were down-regulated in the ADPKD group.Bioinformatics analysis results predicted that significantly up-regulated microRNA were mainly involved in proximal tubule bicarbonate reclamation pathway,glutamate metabolism and pluripotency of stem cells regulating path-ways.Down-regulated microRNA might participate in MAPK signaling pathway and cell adhesion pathway.The expres-sion changes of miR-1291 and miR-6852-5p between groups verified by qRT-PCR were consistent with the se-quencing results(P<0.05).The results of ROC curve showed the miR-1291(AUC=0.73)and miR-6852-5p(AUC=0.84)may have the potential to discriminate between ADPKD and SRC.Combined testing of two indicators may

关 键 词：常染色体显性多囊肾病 外泌体 微小RNA 高通量测序 鉴别诊断 

分 类 号：R692[医药卫生—泌尿科学]