[1]陆 健,冯 萍,吴 静,等.类风湿关节炎活动期患者血浆外泌体miRNAs 差异表达筛选与生物信息学分析及验证[J].现代检验医学杂志,2024,39(02):62-67.[doi:10.3969/j.issn.1671-7414.2024.02.012]
　LU Jian,FENG Ping,WU Jing,et al.Bioinformatics Analysis and Validation of Differential Expression of miRNAs in Plasma Exosomes from Patients with Active Rheumatoid Arthritis[J].Journal of Modern Laboratory Medicine,2024,39(02):62-67.[doi:10.3969/j.issn.1671-7414.2024.02.012]
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类风湿关节炎活动期患者血浆外泌体miRNAs 差异表达筛选与生物信息学分析及验证()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第39卷
期数:
2024年02期
页码:
62-67
栏目:
论著
出版日期:
2024-03-15

文章信息/Info

Title:
Bioinformatics Analysis and Validation of Differential Expression of miRNAs in Plasma Exosomes from Patients with Active Rheumatoid Arthritis
文章编号:
1671-7414（2024）02-062-06
作者:
陆 健1; 冯 萍1; 吴 静2; 杨 欢1
（1. 苏州大学附属第二医院，江苏苏州 215004；2. 苏州市广济医院，江苏苏州，215137）
Author(s):
LU Jian1;  FENG Ping1;  WU Jing2;  YANG Huan1
（1. the Second Affiliated Hospital of Soochow University, Jiangsu Suzhou 215004, China; 2. Suzhou Guangji Hospital, Jiangsu Suzhou 215137, China）
关键词:
类风湿关节炎; 外泌体; 微小核糖核酸; 生物信息学分析
分类号:
R593.22；R392.11
DOI:
10.3969/j.issn.1671-7414.2024.02.012
文献标志码:
A
摘要:
目的　筛选类风湿关节炎（rheumatoid arthritis，RA）活动期患者和健康体检者血浆外泌体（exosomes）中差异表达的微小核糖核酸（microRNAs，miRNAs），并进行生物信息学分析，以探讨血浆外泌体miRNAs 在RA 致病中的作用及其潜在临床应用价值。方法　选取2023 年01 ～ 04 月就诊于苏州大学附属第二医院风湿免疫科的39 例RA 患者作为研究对象，同时选取39 例健康体检者作为正常对照。利用Illumina 高通量测序技术检测血浆外泌体中miRNAs的表达水平，以log2(Fold Change) 绝对值>1 和P 值<0.05 为条件获得差异表达的miRNAs。按照P 值从小到大的顺序挑选6 条miRNAs 进行生物信息学分析，并利用实时荧光定量PCR（quantitative real-time fluorescence PCR，qRT-PCR）验证。结果　与对照组相比，RA 患者血浆外泌体中存在22 条异常表达的miRNAs，4 条上调，18 条下调。其中，miR-30b-5p，miR-144-3p，miR-20a-5p，miR-223-5p，miR-425-3p 和miR-589-5p 的水平变化显著。GO 和KEGG 富集分析结果显示，差异表达的miRNAs 可能通过调节转化生长因子-β（TGF-β）和PI3K/AKT 信号通路参与疾病进展，涉及Th17 细胞分化、细胞间相互作用和蛋白磷酸化等生物学过程。qRT-PCR 验证结果显示，与对照组相比，RA 患者血浆外泌体miR-144-3p 和miR-425-3p 的表达水平明显降低，差异具有统计学意义（t=3.617，3.595，均P<0.001），而miR-30b-5p，miR-223-5p，miR-589-5p 和miR-20a-5p 的表达差异均无统计学意义（t=1.956，1.331，1.662，1.861，均P>0.05）。结论　RA 患者血浆外泌体的miRNAs 表达谱发生改变，可能通过TGF-β 等信号通路参与疾病进展，外泌体miR-144-3p 和miR-425-3p 是RA 疾病诊断的潜在血清学标志物。
Abstract:
Objective　To screen differentially expressed microRNAs (miRNAs) in plasma exosomes of active rheumatoid arthritis (RA) patients and healthy controls and conduct bioinformatics analysis for exploring the role and potential clinical application value of miRNAs in the pathogenesis of RA. Methods　From January 2023 to April 2023, 39 RA patients who visited the Rheumatology and Immunology Department of the Second Affiliated Hospital of Soochow University were selected as the study subjects, while 39 healthy individuals were selected as normal controls. The expression levels of miRNAs in plasma exosomes were detected by Illumina high-throughput sequencing technology, and the differentially expressed miRNAs were obtained by log2 (Fold Change) absolute value >1 and P value <0.05. Six miRNAs were selected by the order from small to large P-value for bioinformatics analysis and validated using quantitative real-time fluorescence PCR (qRT-PCR). Results　Compared with healthy controls, 22 aberrantly expressed miRNAs were detected in plasma exosomes of RA patients, of which 4 were upregulated and 18 were down-regulated. Among them, miR-30b-5p, miR-144-3p, miR-20a-5p, miR-223-5p, miR-425-3p, and miR-589-5p showed changed significantly. GO and KEGG enrichment analysis indicated that differentially expressed miRNAs may be involved in disease progression through regulation of signaling pathways such as TGF-β and PI3K/AKT, which were related to biological processes such as Th17 differentiation, intercellular interactions, and protein phosphorylation. The qRT-PCR validation results showed that the expression of miR-144-3p and miR-425-3p were significantly reduced in plasma exosomes of RA patients compared to healthy controls (t=3.617, 3.595, all P<0.001), while the differences of miR-30b-5p, miR-223-5p, miR- 589-5p, and miR-20a-5p expression were not statistically significant (t=1.956, 1.331, 1.662, 1.861, all P>0.05). Conclusion　 The expression profile of plasma exosomal miRNAs changed in RA patients, which may be involved in disease progression through TGF-β and other signaling pathways. Exosome-derived miR-144-3p and miR-425-3p may be potential serological markers for RA diagnosis.

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备注/Memo

备注/Memo:
基金项目： 江苏省卫健委医学科研项目（Z 2023043）：类风湿关节炎相关血浆外泌体miR-144-3p 活化B 细胞促进自身抗体产生及其应用价值的研究；江苏省妇幼保健协会科研项目（FYX202123）：血清及血清外泌体来源的hsa_circ_0065214 在乳腺癌中的作用机制及临床应用研究；苏州市科技计划项目（关键技术）（SKY2021045）：血清及血清外泌体来源circSCAP_020 作为乳腺癌新型肿瘤标志物的技术应用研究；苏州大学附属第二医院科研预研基金（SDFEYBS2203）：类风湿性关节炎患者血浆外泌体调控 B 细胞异常分化及其致病机制研究。
作者简介：陆健（1993-），男，博士，主管技师，主要从事临床检验诊断学及医学免疫学，E-mail：ljsdf2y@163.com。
通信作者：杨欢（1982-），女，博士，主任技师，主要从事临床检验诊断学及分子生物学，E-mail：zjyhcherry@126.com。
更新日期/Last Update: 2024-03-15