Zhang Mengyuan, Liu Siyu, Wang Yongheng, Shan Shan, Cang Ming
State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Hohhot 010070, China.
Cells. 2025 Aug 10;14(16):1234. doi: 10.3390/cells14161234.
Fibro-adipogenic progenitor cells (FAPs) support muscle tissue homeostasis, regulate muscle growth, injury repair, and fibrosis, and activate muscle progenitor cell differentiation to promote regeneration. We aimed to investigate the effects of co-culturing FAPs with muscle satellite cells (MuSCs) on myogenic differentiation. Proteomic profiling of co-culture supernatants identified significant DCX, IMP2A, NUDT16L1, SLC38A2, and IL-6 upregulation. Comparative transcriptomics of mono-cultured versus co-cultured MuSCs revealed differential expression of oxidative stress-related genes (, , , , , and ). Pathway enrichment analyses highlighted cell cycle regulation, TNF signaling, and ferroptosis. Gene ontology analysis of MuSCs indicated significant gene enrichment in myosin-related components. Combined transcriptomic and proteomic analyses demonstrated HO-1 downregulation at the transcriptional and translational levels, with altered pathways being predominantly related to myosin filament, muscle system process, and muscle contraction cellular components. HO-1 knockdown reduced intracellular iron accumulation in MuSCs, suppressing iron-dependent autophagy. This alleviated oxidative stress and promoted myogenic differentiation. Exogenous IL-6 (0.1 ng/mL) downregulated HO-1 expression, initiating an identical regulatory cascade, while HO-1 overexpression reversed the IL-6-mediated reduction in the expression of the autophagy markers LC3 and ATG5, suppressing myogenic enhancement. This establishes the co-culture-induced IL-6/HO-1 axis as a core regulator of iron-dependent oxidative stress and autophagy during myogenic differentiation.
成纤维脂肪生成祖细胞（FAPs）维持肌肉组织稳态，调节肌肉生长、损伤修复和纤维化，并激活肌肉祖细胞分化以促进再生。我们旨在研究FAPs与肌肉卫星细胞（MuSCs）共培养对成肌分化的影响。共培养上清液的蛋白质组分析确定了双皮质素（DCX）、IMP2A、NUDT16L1、溶质载体家族38成员2（SLC38A2）和白细胞介素-6（IL-6）的显著上调。单培养与共培养的MuSCs的比较转录组学揭示了氧化应激相关基因（……）的差异表达。通路富集分析突出了细胞周期调控、肿瘤坏死因子（TNF）信号传导和铁死亡。MuSCs的基因本体分析表明肌球蛋白相关成分有显著的基因富集。转录组和蛋白质组联合分析表明血红素加氧酶-1（HO-1）在转录和翻译水平下调，改变的通路主要与肌球蛋白丝、肌肉系统过程和肌肉收缩细胞成分有关。HO-1基因敲低减少了MuSCs中的细胞内铁积累，抑制了铁依赖性自噬。这减轻了氧化应激并促进了成肌分化。外源性IL-6（0.1 ng/mL）下调HO-1表达，启动相同的调控级联反应，而HO-1过表达逆转了IL-6介导的自噬标志物微管相关蛋白1轻链3（LC3）和自噬相关蛋白5（ATG5）表达的降低，抑制了成肌增强。这确立了共培养诱导的IL-6/HO-1轴作为成肌分化过程中铁依赖性氧化应激和自噬的核心调节因子。
