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目的：筛选骨关节炎（osteoarthritis,OA）患者滑膜液外泌体中关键微小核糖核酸（micro ribonucleic acid,miRNAs），研究其靶基因在关节软骨细胞中的表达。方法 ：在基因表达综合数据库（Gene Expression Omnibus,GEO）中检索并下载GSE126677及GSE114007数据集，采用DESeq软件筛选OA患者和正常者滑膜液外泌体及软骨组织中差异miRNAs，通过WebGestalt网站对差异表达的miRNAs进行GO/KEGG富集分析，利用TargetScan软件预测差异表达miRNAs作用的靶分子，并将预测得到的靶分子与芯片中差异miRNAs进行交集分析；在组织样本中利用qRT-PCR、Western blot、荧光素酶报告实验，验证差异miRNAs及其靶分子的表达水平及直接靶向结合关系。结果：通过GSE126677芯片分析，筛选出5个外泌体来源的差异表达miRNAs，其中3个在OA组高表达，2个低表达。对GSE114007数据库中OA患者关节软骨组织的mRNA-seq表达矩阵进行分析，共得到1 743个差异表达基因，其中1 010个基因高表达、733个基因低表达。将外泌体差异miRNAs的潜在靶分子与软骨组织差异基因进行交集分析，最终得到候选靶标。组织样本实验进一步证实滑膜液外泌体中低表达的miR-3196靶向调控软骨细胞内的丝裂原活化蛋白激酶15(mitogen-activated protein kinase 15,MAPK15)的表达。结论：OA患者滑膜液外泌体中低表达的miR-3196通过靶向调控软骨细胞中MAPK15，激活下游信号通路，影响软骨细胞的正常生物学功能，参与OA的发生发展。
Abstract：Objective:To screen the key micro ribonucleic acid(miRNAs) in the exosomes of synovial fluid in the patients with osteoarthritis(OA) and investigate their target genes in articular chondrocytes. Methods: Search and download GSE126677 and GSE114007 in Gene Expression Omnibus(GEO)database. The DESeq software was used to screen for differentially expressed miRNAs in the synovial fluid exosomes and cartilage tissues of OA patients and normal individuals.The GO/KEGG enrichment analysis of differentially expressed miRNAs was conducted through the WebGestalt website. The target molecules of the differentially expressed miRNAs were predicted by using the TargetScan software, and the intersection analysis was performed between the predicted target molecules and the differentially expressed miRNAs in the chip.The expression of the selected differentially expressed miRNAs and their target molecules, as well as their direct target binding relationships, were verified in the tissue samples using qRT-PCR, Western blot, and luciferase reporter assays.Results: Through the analysis of the GSE126677 chip, 5 differentially expressed miRNAs from exosomes were identified.Among them, 3 were highly expressed, and 2 were lowly expressed in the OA group. The mRNA-seq expression matrix of the joint cartilage tissues of OA patients in the GSE114007 database was analyzed. A total of 1743 differentially expressed genes were obtained, among which 1010 genes were highly expressed and 733 genes were lowly expressed. The potential target molecules of the differential miRNAs in exosomes were intersected with the differential genes in cartilage tissue, and the candidate targets were finally obtained. The tissue sample experiments further confirmed that the low expression of miR-3196 in synovial fluid exosomes targeted and regulated the expression of mitogen-activated protein kinase 15(MAPK15) in chondrocytes. Conclusion: The low expression of miR-3196 in the synovial fluid exosomes of OA patients activates downstream signaling pathways by targeting and regulating MAPK15 in chondrocytes, thereby affecting the normal biological functions of chondrocytes and participating in the occurrence and development of OA.
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基本信息:
DOI：10.19767/j.cnki.32-1412.2026.03.002
中图分类号:R684.3
引用信息:
[1]顾思源,周吉秋,冒星星,等.骨关节炎患者滑膜液外泌体关键miRNAs的筛选及其在关节软骨细胞中靶基因研究[J].交通医学,2026,40(03):235-242.DOI:10.19767/j.cnki.32-1412.2026.03.002.
2026-06-20
2026-06-20