Skeletal Muscle Cell Oxidative Stress as a Possible Therapeutic Target in a Denervation-Induced Experimental Sarcopenic Model
Skeletal Muscle Cell Oxidative Stress as a Possible Therapeutic Target in a Denervation-Induced Experimental Sarcopenic Model
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DOI：
10.1097/brs.0000000000002891
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发表时间：
2019-04-15
期刊：
影响因子：
3
通讯作者：
Ohtori, Seiji
中科院分区：
文献类型：
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作者：
Kinoshita, Hideyuki;Orita, Sumihisa;Ohtori, Seiji
研究设计。使用肌肉减少症啮齿动物模型进行基础研究。目的。使用神经源性肌少症动物模型阐明氧化应激对肌肉变性的影响以及抗氧化剂治疗肌少症的功效。背景数据摘要。据报道，氧化应激与许多病理学有关，包括肌肉骨骼疾病。然而，它与肌肉减少症（腰痛的潜在根源之一）的关系尚未完全了解。方法。成肌细胞系 (C2C12) 用氧化应激诱导剂 H2O2 和抗氧化剂 N-乙酰基-L-半胱氨酸 (NAC) 处理。通过蛋白质印迹、免疫细胞化学和 3-(4,5-二甲基噻唑-2-基)2,5-二苯基溴化四唑 (MTT) 细胞活力测定来评估氧化应激诱导的细胞凋亡作用和 NAC 的抗氧化作用。通过坐骨神经轴切术产生肌肉减少症动物模型以诱导肌肉萎缩。将二十四只雄性 Sprague-Dawley 大鼠分为假手术组、shamthornNAC 组、轴切术组和轴切术组。给大鼠提供仅水或含有 NAC (1 g/L) 的水 4 周。轴索切除术后2周，分离腓肠肌并用苏木精和伊红(H&E)染色，从中收获肌细胞并提取蛋白质用于评估。结果。在 C2C12 细胞中，H2O2 处理显着激活丝裂原激活蛋白激酶 (MAPK)，NAC 预处理可改善这种情况。此外，H2O2 诱导 C2C12 细胞凋亡和死亡，而 NAC 预处理可以阻止这种情况。轴突切断术组的腓肠肌重量减轻，而 NAC 给药可以防止这种情况发生。最后，虽然轴突切除组的肌肉标本显示肌纤维减少较多，但口服 NAC 通过抗氧化作用显着抑制肌萎缩。结论。目前的体外和体内研究表明氧化应激可能参与肌肉减少症的病理学。 NAC 是一种潜在的抗肌肉减少症候选药物，可预防肌萎缩和脂肪变性。
Study Design. A basic study using a rodent model of sarcopenia.Objective. To elucidate the contribution of oxidative stress to muscle degeneration and the efficacy of antioxidant treatment for sarcopenia using an animal model of neurogenic sarcopenia.Summary of Background Data. Oxidative stress has been reported to be involved in a number of pathologies, including musculoskeletal disorders. Its relationship with sarcopenia, one of the potential origins of lower back pain, however, is not yet fully understood.Methods. Myoblast cell lines (C2C12) were treated with H2O2, an oxidative stress inducer, and N-acetyl-L-cysteine (NAC), an antioxidant. Apoptotic effects induced by oxidative stress and the antioxidant effects of NAC were assessed by western blotting, immunocytochemistry, and 3-(4,5-dimethylthiazol-2-yl)2,5- diphenyltetrazolium bromide (MTT) cell viability assays. An animal model of sarcopenia was produced via axotomy of the sciatic nerves to induce muscle atrophy. Twenty-four male Sprague-Dawley rats were divided into sham, shamthornNAC, axotomy, and axotomythornNAC groups. Rats were provided water only or water containing NAC (1 g/L) for 4 weeks. The gastrocnemius muscle was isolated and stained with hematoxylin and eosin (H& E) 2 weeks after axotomy, from which muscle cells were harvested and protein extracted for evaluation.Results. Mitogen-activated protein kinases (MAPKs) were significantly activated by H2O2 treatment in C2C12 cells, which was ameliorated by NAC pretreatment. Furthermore, H2O2 induced apoptosis and death of C2C12 cells, which was prevented by NAC pretreatment. The weight of the gastrocnemius muscle was reduced in the axotomy group, which was prevented by NAC administration. Lastly, although muscle specimens from the axotomy group showed greater reductions in muscle fiber, the oral administration of NAC significantly inhibited amyotrophy via antioxidant effects.Conclusion. The current in vitro and in vivo study demonstrated the possible involvement of oxidative stress in sarcopenic pathology. NAC represents a potential anti-sarcopenic drug candidate, preventing amyotrophy and fatty degeneration.