肿瘤坏死因子α（TNFα）具有多种生物学功能；然而，其在肌生成中的调节作用尚不完全清楚。在本研究中，我们探讨了 TNFα 在原代成肌细胞和 C2C12 细胞中成肌细胞增殖、分化、迁移和肌管融合中的功能。为此，我们构建了TNFα肌肉条件敲除（TNFα-CKO）小鼠，并将其与flox小鼠进行比较，以评估TNFα敲除对骨骼肌的影响。结果表明，与flox小鼠相比，TNFα-CKO小鼠表现出肌肉发育加速、再生能力增强、运动耐力提高等表型，但主要内脏器官或骨骼结构没有观察到显着差异。使用无标记蛋白质组学分析，我们发现 TNFα-CKO 改变了几种肌肉发育相关蛋白在细胞核和细胞质中的分布，例如 Hira、Casz1、Casp7、Arhgap10、Gas1、Diaph1、Map3k20、Cfl2 和 Igf2 。基因集富集分析 (GSEA) 进一步揭示，TNFα 缺陷导致氧化磷酸化和 MyoD 靶标呈正富集，而 JAK-STAT 信号传导呈负富集。这些发现表明 TNFα-CKO 可能通过这些新确定的靶点和途径积极调节肌肉生长和发育。

Tumor necrosis factor α (TNFα) exhibits diverse biological functions; however, its regulatory roles in myogenesis are not fully understood. In the present study, we explored the function of TNFα in myoblast proliferation, differentiation, migration, and myotube fusion in primary myoblasts and C2C12 cells. To this end, we constructed TNFα muscle-conditional knockout ( TNFα-CKO) mice and compared them with flox mice to assess the effects of TNFα knockout on skeletal muscles. Results indicated that TNFα-CKO mice displayed phenotypes such as accelerated muscle development, enhanced regenerative capacity, and improved exercise endurance compared to flox mice, with no significant differences observed in major visceral organs or skeletal structure. Using label-free proteomic analysis, we found that TNFα-CKO altered the distribution of several muscle development-related proteins, such as Hira, Casz1, Casp7, Arhgap10, Gas1, Diaph1, Map3k20, Cfl2, and Igf2, in the nucleus and cytoplasm. Gene set enrichment analysis (GSEA) further revealed that TNFα deficiency resulted in positive enrichment in oxidative phosphorylation and MyoD targets and negative enrichment in JAK-STAT signaling. These findings suggest that TNFα-CKO positively regulates muscle growth and development, possibly via these newly identified targets and pathways.