Ex Vivo Equine Cartilage Explant Osteoarthritis Model: A Metabolomics and Proteomics Study
Ex Vivo Equine Cartilage Explant Osteoarthritis Model: A Metabolomics and Proteomics Study
复制标题
DOI：
10.1021/acs.jproteome.0c00143
复制
发表时间：
2020-09-04
影响因子：
4.4
通讯作者：
Peffers, Mandy J.
中科院分区：
文献类型：
--
作者：
Anderson, James R.;Phelan, Marie M.;Peffers, Mandy J.
骨关节炎是一种与年龄相关的退行性肌肉骨骼疾病，其特征是关节软骨丧失、滑膜炎和软骨下骨硬化。骨关节炎的发病机制尚未完全阐明，临床上没有骨关节炎特异性的生物标志物。体外马软骨外植体（n = 5）在补充肿瘤坏死因子- α (tnf - α)/白细胞介素-1 β (IL-1 β)的培养基中培养8天，分别于2、5和8天取出并更换培养基。8天软骨外植体和培养基样品的乙腈代谢物提取在所有时间点进行一维（1D） H-1核磁共振代谢组学分析，培养基样品也进行质谱蛋白质组学分析。在软骨内，tnf - α /IL-1 β治疗后，葡萄糖和赖氨酸升高，而腺苷、丙氨酸、甜菜碱、肌酸、肌醇和尿苷降低。在培养基中，分别在1-2、3-5和6-8天鉴定出4、4和6种差异丰富的代谢物，以及154、138和72种差异丰富的蛋白质，包括处理后丙氨酸减少，异亮氨酸、烯醇化酶1、vimentin和lamin A/C增加。9种潜在的新型骨关节炎新肽在治疗介质中升高。受牵连的通路主要是那些参与细胞运动的通路。我们的创新研究为早期骨关节炎的发病机制提供了深刻的信息，为临床标志物和可能的新治疗靶点提供了潜在的翻译。
Osteoarthritis is an age-related degenerative musculoskeletal disease characterized by loss of articular cartilage, synovitis, and subchondral bone sclerosis. Osteoarthritis pathogenesis is yet to be fully elucidated with no osteoarthritis-specific biomarkers in clinical use. Ex vivo equine cartilage explants (n = 5) were incubated in tumor necrosis factor-alpha (TNF-alpha)/interleukin-1 beta (IL-1 beta)-supplemented culture media for 8 days, with the media removed and replaced at 2, 5, and 8 days. Acetonitrile metabolite extractions of 8 day cartilage explants and media samples at all time points underwent one-dimensional (1D) H-1 nuclear magnetic resonance metabolomic analysis, with media samples also undergoing mass spectrometry proteomic analysis. Within the cartilage, glucose and lysine were elevated following TNF-alpha/IL-1 beta treatment, while adenosine, alanine, betaine, creatine, myo-inositol, and uridine decreased. Within the culture media, 4, 4, and 6 differentially abundant metabolites and 154, 138, and 72 differentially abundant proteins were identified at 1-2, 3-5, and 6-8 days, respectively, including reduced alanine and increased isoleucine, enolase 1, vimentin, and lamin A/C following treatment. Nine potential novel osteoarthritis neopeptides were elevated in the treated media. Implicated pathways were dominated by those involved in cellular movement. Our innovative study has provided insightful information on early osteoarthritis pathogenesis, enabling potential translation for clinical markers and possible new therapeutic targets.