ABSTRACT

The current study aimed to explored the regulatory effect of Tropomyosin-related kinases B (TrkB) in the development and function of chondrocyte. Correlation between clinicopathological characteristics and osteoarthritis (OA) were analyzed. The expressions of TrkA, brain-derived neurotrophic factor (BDNF), TrkB, Src homolog and collagen homolog B (ShcB), and ShcC in OA cartilage tissue and IL-1β-stimulated chondrocytes from normal cartilage were determined by Western blot/qRT-PCR. After manipulating the expressions of TrkA, shTrkB, ShcB, miR-146a-3p and nuclear paraspeckle assembly transcript 1 (NEAT1), the differentiation-related molecules, and apoptosis-related molecules were examined by Western blot/qRT-PCR, and migration, invasion, proliferation, tube formation, and apoptosis rate in IL-1β-stimulated chondrocyte were examined by scratch, Transwell, colony formation, and tube formation, and flow cytometry assays, respectively. Bioinformatics, dual-luciferase and Spearman were used to analyze the binding and correlation of target genes. The findings showed that OA was related to body mass Index (BMI). The expressions of TrkA, TrkB and ShcB and NEAT1 were up-regulated in OA and IL-1β-stimulated chondrocytes, while miR-146a-3p was donwnregulated and was negatively correlated with TrkB or NEAT1. NEAT1 competed with TrkB in chondrocytes for miR-146a-3p binding. ShTrkB reversed the decrease in expressions of differentiation-related molecules, migration, invasion and proliferation, and the increase in ShcB expression and tube formation, of IL-1β-stimulated chondrocytes. Overexpressed ShcB reversed effect of shTrkB on the functions of IL-1β-stimulated chondrocytes. MiR-146a-3p inhibitor reversed effects of shTrkB on the function and apoptosis-related molecules on IL-1β-stimulated chondrocytes, while NEAT1 reversed role of miR-146a-3p. This paper demonstrated that NEAT1/miR-146a-3p/TrkB/ShcB axis regulates the development and function of chondrocyte.

 抽象的

本研究旨在探讨原肌球蛋白相关激酶B（TrkB）对软骨细胞发育和功能的调节作用。分析临床病理特征与骨关节炎（OA）之间的相关性。通过 Western blot/qRT 测定 OA 软骨组织和正常软骨中 IL-1β 刺激的软骨细胞中 TrkA、脑源性神经营养因子 (BDNF)、TrkB、Src 同源物和胶原同源物 B (ShcB) 和 ShcC 的表达。 PCR。操纵TrkA、shTrkB、ShcB、miR-146a-3p和核副斑组装转录物1（NEAT1）的表达后，通过Western blot/qRT-PCR和迁移检查分化相关分子和凋亡相关分子，分别通过划痕、Transwell、集落形成和管形成以及流式细胞术检测IL-1β刺激的软骨细胞的侵袭、增殖、管形成和凋亡率。利用生物信息学、双荧光素酶和Spearman分析靶基因的结合和相关性。研究结果表明，OA与体重指数（BMI）相关。 OA和IL-1β刺激的软骨细胞中TrkA、TrkB、ShcB和NEAT1的表达上调，而miR-146a-3p下调且与TrkB或NEAT1呈负相关。 NEAT1 与软骨细胞中的 TrkB 竞争 miR-146a-3p 结合。 ShTrkB 逆转了 IL-1β 刺激的软骨细胞中分化相关分子表达、迁移、侵袭和增殖的减少，以及 ShcB 表达和管形成的增加。过表达的 ShcB 逆转了 shTrkB 对 IL-1β 刺激的软骨细胞功能的影响。 MiR-146a-3p抑制剂逆转了shTrkB对IL-1β刺激的软骨细胞的功能和凋亡相关分子的影响，而NEAT1则逆转了miR-146a-3p的作用。该论文证明NEAT1/miR-146a-3p/TrkB/ShcB轴调控软骨细胞的发育和功能。