Walters Marie, Skovgaard Kerstin, Heegaard Peter M H, Fang Yongxiang, Kharaz Yalda A, Bundgaard Louise, Skovgaard Lene T, Jensen Henrik E, Andersen Pia H, Peffers Mandy J, Jacobsen Stine
Department of Veterinary Clinical Sciences, University of Copenhagen, Taastrup, Denmark.
Department of Biotechnology and Biomedicine, Technical University of Denmark, Kgs. Lyngby, Denmark.
Equine Vet J. 2025 Jul;57(4):1138-1150. doi: 10.1111/evj.14456. Epub 2025 Jan 8.
MicroRNAs, a class of small noncoding RNAs, serve as post-transcriptional regulators of gene expression and are present in a stable and quantifiable form in biological fluids. MicroRNAs may influence intra-articular responses and the course of disease, but very little is known about their temporal changes in osteoarthritis.
To identify miRNAs and characterise the temporal changes in their abundance in SF from horses with experimentally induced osteoarthritis. We hypothesised that the abundance of miRNA would change during disease progression.
In vivo experiments.
RNA extracted from synovial fluid obtained sequentially (Day 0, 28 and 70) from nine horses with experimentally induced osteoarthritis was subjected to small RNA sequencing using the Illumina Hiseq 4000 sequencing platform. Differentially abundant miRNAs underwent further validation and mapping of temporal abundance (Day 0, 14, 17, 21, 28, 35, 42, 49, 56, 63 and 70 days after osteoarthritis induction) by microfluidic reverse transcription quantitative real-time PCR. Bioinformatic analyses were performed to predict potential biological associations and target genes of the differentially abundant microRNAs.
Small RNA sequencing revealed 61 differentially abundant microRNAs at an early osteoarthritis stage (Day 28), and subsequent reverse transcription quantitative real-time PCR analysis validated 20 of these. Significant biological functions of the differentially abundant microRNAs were apoptosis, necrosis, cell proliferation and cell invasion. Following validation, four microRNAs (miRNA-199b-3p, miRNA-139-5p, miRNA-1839 and miRNA-151-5p) were detected in more than 50% of the synovial fluid samples and had higher abundance in osteoarthritic than in control joints.
Limited sample size.
This is the first study to determine longitudinal changes in synovial fluid microRNA abundance in an equine model of osteoarthritis. Larger studies are needed in naturally occurring osteoarthritis to interrogate putative changes identified by this study.
微小RNA是一类小的非编码RNA，作为基因表达的转录后调节因子，以稳定且可量化的形式存在于生物体液中。微小RNA可能影响关节内反应和疾病进程，但对于它们在骨关节炎中的时间变化知之甚少。
鉴定微小RNA并表征实验性诱导骨关节炎马匹滑液中其丰度的时间变化。我们假设微小RNA的丰度会在疾病进展过程中发生变化。
体内实验。
从9匹实验性诱导骨关节炎的马匹中依次（第0、28和70天）获取的滑液中提取RNA，使用Illumina Hiseq 4000测序平台进行小RNA测序。对差异丰度的微小RNA进行进一步验证，并通过微流控逆转录定量实时PCR绘制时间丰度图（骨关节炎诱导后第0、14、17、21、28、35、42、49、56、63和70天）。进行生物信息学分析以预测差异丰度微小RNA的潜在生物学关联和靶基因。
小RNA测序在骨关节炎早期阶段（第28天）揭示了61种差异丰度的微小RNA，随后的逆转录定量实时PCR分析验证了其中20种。差异丰度微小RNA的重要生物学功能是细胞凋亡、坏死、细胞增殖和细胞侵袭。验证后，在超过50%的滑液样本中检测到4种微小RNA（miRNA-199b-3p、miRNA-139-5p、miRNA-1839和miRNA-151-5p），并且它们在骨关节炎关节中的丰度高于对照关节。
样本量有限。
这是第一项确定骨关节炎马模型中滑液微小RNA丰度纵向变化的研究。需要在自然发生的骨关节炎中进行更大规模的研究，以探究本研究确定的假定变化。
