滑膜成纤维细胞来源的 miR-214-3p 外泌体抑制骨关节炎大鼠软骨组织的炎症和退变。
Synovial fibroblast-miR-214-3p-derived exosomes inhibit inflammation and degeneration of cartilage tissues of osteoarthritis rats.
机构信息
Department of Orthopedics, Jinling Hospital, Nanjing University, School of Medicine, No. 305 East Zhongshan Road, Nanjing, 210002, China.
Department of Orthopedics, The People's Hospital of Wugang City, Wugang, 422400, China.
出版信息
Mol Cell Biochem. 2023 Mar;478(3):637-649. doi: 10.1007/s11010-022-04535-9. Epub 2022 Aug 24.
MicroRNAs (miRs) are regulators of number of cellular process. miRs enclosed within exosomes can be crucial regulators of intercellular signalling and could be an important biomarker of various age-associated disorders. Role of exosomal enclosed miRs in osteoarthritis (OA) chondrocytes and synovial fibroblasts (SFBs) remains poorly studied. Here, we profiled and studied the effect of synovial fluid-derived exosomal miRs on inflammation, survival, proliferation of chondrocyte in correlation with cartilage degeneration. Exosomes were isolated from synovial fluid collected from OA subjects and were analysed by transmission electron microscopy. miRs were isolated and were submitted to microarray profiling. Web-based PCR analysis was done. Chondrocyte proliferation and colony formation assay were performed. Apoptosis study was done by flow cytometer. Gene expression was done by qRT-PCR analysis and protein expression by western blot assay. Rat model of OA was created by operating the knee by anterior cruciate ligament and resection of medial menisci (ACLT + MMx) method. Micro-CT analysis, histological analysis, immunohistochemical staining, and TUNEL assay were also performed. About 17 miRs were found to be expressed differentially in the synovial fluid collected from the control and OA subjects. Microarray analysis confirmed, expression of miR-214-3p was significantly downregulated in the synovial fluid exosome of OA subjects. miR-214-3p mimic promoted proliferation of chondrocyte and suppressed apoptosis. Treatment also inhibited the levels of TNF-α, IL-1β and IL-6. SFB-miR-214-3p exosomes suppressed apoptosis and also inflammation in chondrocytes. In vivo study suggested that SFB-exosomal miR-214-3p from rats suppressed the formation of osteophytes, prevented degeneration of cartilage and exerted anti-inflammatory and anti-apoptotic effect in articular cartilage tissue. The findings suggested that SFB-miR-214-3p exosomes can ameliorate chondrocyte inflammation and degeneration of cartilage tissues. The study confirms therapeutic potential of SFB-miR-214-3p exosomes in treating OA.
微小 RNA(miRs)是许多细胞过程的调节剂。包含在细胞外体中的 miRs 可能是细胞间信号的重要调节剂,并且可以作为各种与年龄相关疾病的重要生物标志物。细胞外体包裹的 miRs 在骨关节炎(OA)软骨细胞和滑膜成纤维细胞(SFB)中的作用仍未得到充分研究。在这里,我们对滑膜液衍生的细胞外体 miRs 对软骨细胞炎症、存活和增殖与软骨退变的相关性进行了分析和研究。通过透射电子显微镜分析分离 OA 患者滑膜液中的外泌体。分离微小 RNA 并进行微阵列分析。进行基于网络的 PCR 分析。进行软骨细胞增殖和集落形成试验。通过流式细胞仪进行细胞凋亡研究。通过 qRT-PCR 分析进行基因表达,通过 Western blot 分析进行蛋白表达。通过前交叉韧带和内侧半月板切除术(ACLT+MMx)方法对大鼠模型进行 OA 操作。还进行了 micro-CT 分析、组织学分析、免疫组织化学染色和 TUNEL 分析。在对照组和 OA 组患者滑膜液中发现约 17 个微小 RNA 表达差异。微阵列分析证实,OA 患者滑膜液外泌体中 miR-214-3p 的表达显著下调。miR-214-3p 模拟物促进软骨细胞增殖并抑制凋亡。治疗还抑制了 TNF-α、IL-1β 和 IL-6 的水平。SFB-miR-214-3p 外泌体抑制软骨细胞凋亡和炎症。体内研究表明,大鼠 SFB 细胞外体 miR-214-3p 抑制骨赘形成,防止软骨退变,并在关节软骨组织中发挥抗炎和抗凋亡作用。研究结果表明,SFB-miR-214-3p 外泌体可改善软骨细胞炎症和软骨组织退变。该研究证实了 SFB-miR-214-3p 外泌体在治疗 OA 中的治疗潜力。