NEAT1/miR-146a-3p/TrkB/ShcB axis regulates the development and function of chondrocyte
NEAT1/miR-146a-3p/TrkB/ShcB axis regulates the development and function of chondrocyte
复制标题
DOI：
10.1080/15384101.2021.1974787
复制
发表时间：
2021-09
期刊：
影响因子：
4.3
通讯作者：
Fanyou Ning;Shaobo Zhu;Hui Gao;Yu Deng
中科院分区：
文献类型：
--
作者：
Fanyou Ning;Shaobo Zhu;Hui Gao;Yu Deng
本研究旨在探讨原肌球蛋白相关激酶B（Trk B）对软骨细胞发育和功能的调控作用。分析临床病理特征与骨关节炎（OA）的相关性。通过Western印迹/qRT-PCR检测OA软骨组织和IL-1β刺激的正常软骨软骨细胞中TrkA、脑源性神经营养因子（BDNF）、Trk B、Src同源物和胶原同源物B（Shc B）以及ShcC的表达。在调控TrkA、shTrkB、ShcB、miR-146 a-3 p和核旁斑组装转录本1（NEAT 1）表达后，通过Western blot/qRT-PCR检测分化相关分子和凋亡相关分子，并通过划痕、Transwell、集落形成和管形成检测IL-1β刺激的软骨细胞的迁移、侵袭、增殖、管形成和凋亡率。和流式细胞术测定。应用生物信息学、双荧光素酶和斯皮尔曼分析方法分析靶基因的结合和相关性。结果表明，OA与体重指数（BMI）有关。在OA和IL-1β刺激的软骨细胞中，TrkA、TrkB、ShcB和NEAT 1的表达上调，而miR-146 a-3 p的表达下调，且与TrkB和NEAT 1呈负相关。NEAT 1与TrkB在软骨细胞中竞争miR-146 a-3 p结合。ShTrkB可逆转IL-1β刺激后软骨细胞分化相关分子表达减少、迁移、侵袭和增殖能力下降以及ShcB表达增加和小管形成增加。过表达ShcB可逆转shTrkB对IL-1β刺激的软骨细胞功能的影响。miR-146 a-3 p抑制剂逆转了shTrkB对IL-1β刺激的软骨细胞功能和凋亡相关分子的影响，而NEAT 1逆转了miR-146 a-3 p的作用。本研究证实NEAT 1/miR-146 a-3 p/TrkB/ShcB轴调控软骨细胞的发育和功能。
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.