TNF-α activates RELA expression via TNFRSF1B to upregulate OPA1 expression and inhibit chondrogenic differentiation of human adipose stem cells.
TNF-α activates RELA expression via TNFRSF1B to upregulate OPA1 expression and inhibit chondrogenic differentiation of human adipose stem cells.
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DOI：
10.1186/s13018-023-03846-x
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发表时间：
2023-06-13
影响因子：
2.6
通讯作者：
Yang, Zhen
中科院分区：
文献类型：
--
作者：
Guo, Jiajia;Ye, Wang;Wu, Xinglin;Huang, Haifeng;Li, Bo;Sun, Zeyu;Ren, Zhijing;Yang, Zhen
关键词：
肿瘤坏死因子-α（TNF-α）是介导关节局部炎症过程的促炎细胞因子之一，其抑制软骨形成并对用于治疗骨关节炎（OA）的基于干细胞的软骨再生具有不利影响。然而，这种抑制作用背后的机制仍然知之甚少。由线粒体融合和分裂介导的线粒体形态变化具有高度可塑性，对环境刺激非常敏感，在维持细胞结构和功能方面起着至关重要的作用。本研究以人脂肪干细胞（human adipose stem cells，hADSCs）为研究对象，观察TNF-α对hADSCs向软骨细胞分化能力及线粒体融合与分裂的影响。目的：探讨正常条件下和TNF-α诱导下，线粒体融合和分裂调控在hADSCs向软骨细胞分化中的作用及机制。采用流式细胞仪检测hADSCs的免疫表型CD 29、CD 44、CD 34、CD 45和HLA-DR，采用Alcian蓝染色和天狼星红染色分别观察hADSCs向软骨细胞分化过程中蛋白多糖和胶原的形成。软骨形成标志物SOX 9、II型胶原（COL 2A 1）和聚集蛋白聚糖的mRNA和蛋白表达水平分别通过实时荧光定量PCR（RT-qPCR）和蛋白质印迹法测量。荧光探针MitoTracker® Red CMXRos和JC-1用于可视化线粒体形态并检测线粒体膜电（MMP）。使用Affyssin PrimeView™芯片进行基因表达谱分析。结果表明，TNF-α抑制hADSCs向软骨细胞分化，并使视神经萎缩蛋白1（optic atrophy 1，OPA 1）表达显著上调，线粒体延长，相互连接。基因芯片和RT-qPCR结果显示，TNF-α诱导hADSCs向软骨细胞分化过程中，TNF-α受体2（TNFRSF 1B）和RELA的表达增加。TNF-α通过TNFRSF 1B上调OPA 1表达激活RELA表达，从而增加线粒体融合，从而抑制人脂肪干细胞的软骨分化。
Tumor necrosis factor-alpha (TNF-α), one of the pro-inflammatory cytokines mediating the local inflammatory process in joints, inhibits cartilage formation and has a detrimental effect on stem cell-based cartilage regeneration for the treatment of osteoarthritis (OA). However, the mechanisms behind this inhibitory effect are still poorly understood. Mitochondrial morphological changes mediated by mitochondrial fusion and fission are highly plastic, are quite sensitive to environmental stimuli and play a crucial role in maintaining cell structure and function. In our study, chondrogenic differentiated human adipose stem cells (hADSCs) were exposed to TNF-α and the effect of TNF-α on the ability of hADSCs to chondrogenic differentiate and on mitochondrial fusion and fission was observed and analyzed. The aim was to investigate the role and mechanisms of mitochondrial fusion and fission regulation in the chondrogenic differentiation of hADSCs under normal conditions and under exposure to TNF-α. We used flow cytometry to identify hADSCs immunophenotypes CD29, CD44, CD34, CD45, and HLA-DR. Alcian blue staining and Sirius red staining were used to observe the formation of proteoglycans and collagen during the chondrogenic differentiation of hADSCs, respectively. The mRNA and protein expression levels of the cartilage formation marker SOX9, type II collagen (COL2A1), and Aggrecan were measured by real-time fluorescent quantitative PCR (RT-qPCR) and western blot, respectively. The fluorescent probes MitoTracker® Red CMXRos and JC-1 were used to visualize mitochondria morphology and detect mitochondrial membrane electricity (MMP). Affymetrix PrimeView™ chips were used for gene expression profiling. The results showed that the chondrogenic differentiation of hADSCs was inhibited in the presence of TNF-α that optic atrophy 1 (OPA1) expression was significantly upregulated and mitochondria were prolonged and interconnected during this process. Gene microarray and RT-qPCR data showed that the presence of TNF-α led to increased expression of TNFα receptor 2 (TNFRSF1B) and RELA during chondrogenic differentiation of hADSCs. TNF-α inhibits chondrogenic differentiation of human adipose stem cells by activating RELA expression through TNFRSF1B upregulating OPA1 expression thereby increasing mitochondrial fusion.
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影响因子：
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