Microsurgery Department of Orthopaedic Center, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, 830054, Xinjiang, China.
Key Laboratory of Trauma Repair and Reconstruction of Xinjiang Province, Xinjiang Medical University, Urumqi, 830054, Xinjiang, China.
J Transl Med. 2024 Nov 2;22(1):991. doi: 10.1186/s12967-024-05810-7.
Muscle atrophy caused by denervation is common in neuromuscular diseases, leading to loss of muscle mass and function. However, a comprehensive understanding of the overall molecular network changes during muscle denervation atrophy is still deficient, hindering the development of effective treatments.
In this study, a sciatic nerve transection model was employed in male C57BL/6 J mice to induce muscle denervation atrophy. Gastrocnemius muscles were harvested at 3 days, 2 weeks, and 4 weeks post-denervation for transcriptomic and proteomic analysis. An integrative multi-omics approach was utilized to identify key genes essential for disease progression. Targeted proteomics using PRM was then employed to validate the differential expression of central genes. Combine single-nucleus sequencing results to observe the expression levels of PRM-validated genes in different cell types within muscle tissue.Through upstream regulatory analysis, NRF2 was identified as a potential therapeutic target. The therapeutic potential of the NRF2-targeting drug Omaveloxolone was evaluated in the mouse model.
This research examined the temporal alterations in transcripts and proteins during muscle atrophy subsequent to denervation. A comprehensive analysis identified 54,534 transcripts and 3,218 proteins, of which 23,282 transcripts and 1,852 proteins exhibited statistically significant changes at 3 days, 2 weeks, and 4 weeks post-denervation. Utilizing multi-omics approaches, 30 hubgenes were selected, and PRM validation confirmed significant expression variances in 23 genes. The findings highlighted the involvement of mitochondrial dysfunction, oxidative stress, and metabolic disturbances in the pathogenesis of muscle atrophy, with a pronounced impact on type II muscle fibers, particularly type IIb fibers. The potential therapeutic benefits of Omaveloxolone in mitigating oxidative stress and preserving mitochondrial morphology were confirmed, thereby presenting novel strategies for addressing muscle atrophy induced by denervation. GSEA analysis results show that Autophagy, glutathione metabolism, and PPAR signaling pathways are significantly upregulated, while inflammation-related and neurodegenerative disease-related pathways are significantly inhibited in the Omaveloxolone group.GSR expression and the GSH/GSSG ratio were significantly higher in the Omaveloxolone group compared to the control group, while MuSK expression was significantly lower than in the control group.
In our study, we revealed the crucial role of oxidative stress, glucose metabolism, and mitochondrial dysfunction in denervation-induced muscle atrophy, identifying NRF2 as a potential therapeutic target. Omaveloxolone was shown to stabilize mitochondrial function, enhance antioxidant capacity, and protect neuromuscular junctions, thereby offering promising therapeutic potential for treating denervation-induced muscle atrophy.
失神经支配引起的肌肉萎缩在神经肌肉疾病中很常见，导致肌肉质量和功能丧失。然而，对于失神经支配萎缩过程中整体分子网络变化的全面了解仍然不足，这阻碍了有效治疗方法的发展。
本研究采用雄性 C57BL/6J 小鼠坐骨神经横断模型诱导肌肉失神经萎缩。在神经切断后 3 天、2 周和 4 周采集比目鱼肌进行转录组和蛋白质组分析。采用整合的多组学方法鉴定疾病进展所必需的关键基因。然后使用 PRM 靶向蛋白质组学验证中心基因的差异表达。结合单细胞测序结果观察 PRM 验证基因在肌肉组织内不同细胞类型中的表达水平。通过上游调控分析，鉴定 NRF2 为潜在的治疗靶点。在小鼠模型中评估 NRF2 靶向药物 Omaveloxolone 的治疗潜力。
本研究检测了失神经支配后肌肉萎缩过程中转录本和蛋白质的时间变化。全面分析鉴定出 54534 个转录本和 3218 个蛋白质，其中 23282 个转录本和 1852 个蛋白质在失神经支配后 3 天、2 周和 4 周时表现出统计学显著变化。利用多组学方法，选择了 30 个枢纽基因，PRM 验证证实了 23 个基因的显著表达差异。研究结果强调了线粒体功能障碍、氧化应激和代谢紊乱在肌肉萎缩发病机制中的作用，对 II 型肌肉纤维，特别是 IIb 型纤维有显著影响。Omaveloxolone 减轻氧化应激和维持线粒体形态的潜在治疗益处得到了证实，从而为治疗失神经支配引起的肌肉萎缩提供了新的策略。GSEA 分析结果表明，自噬、谷胱甘肽代谢和 PPAR 信号通路显著上调，而炎症相关和神经退行性疾病相关通路显著受到抑制。与对照组相比，奥马伐罗酮组的 GSR 表达和 GSH/GSSG 比值显著升高，而 MuSK 表达显著低于对照组。
在本研究中，我们揭示了氧化应激、葡萄糖代谢和线粒体功能障碍在失神经支配引起的肌肉萎缩中的关键作用，鉴定 NRF2 为潜在的治疗靶点。奥马伐罗酮可稳定线粒体功能，增强抗氧化能力，保护神经肌肉接头，为治疗失神经支配引起的肌肉萎缩提供了有前景的治疗潜力。
