Elastase levels and activity are increased in dystrophic muscle and impair myoblast cell survival, proliferation and differentiation.
Elastase levels and activity are increased in dystrophic muscle and impair myoblast cell survival, proliferation and differentiation.
复制标题
DOI：
10.1038/srep24708
复制
发表时间：
2016-05-31
影响因子：
4.6
通讯作者：
Pisconti A
中科院分区：
文献类型：
--
作者：
Arecco N;Clarke CJ;Jones FK;Simpson DM;Mason D;Beynon RJ;Pisconti A
在杜氏肌营养不良症中，肌肉组织的进行性丧失伴随着纤维化、慢性炎症以及肌肉再生能力的降低。尽管对于肌营养不良症中纤维化和慢性炎症的发展已有很多了解，但对于它们如何在机制上与肌肉再生能力的丧失相关联却知之甚少。我们开发了一种蛋白质组学方法来发现肌肉祖细胞微环境中与营养不良相关的变化，该方法确定了丝氨酸蛋白酶，尤其是中性粒细胞弹性蛋白酶作为候选因素。我们表明，弹性蛋白酶活性在营养不良（mdx4cv）肌肉中增加，并损害培养中的成肌细胞存活。虽然弹性蛋白酶对C2C12细胞存活的影响与弹性蛋白酶介导的细胞所附着底物的降解动力学相关，但弹性蛋白酶对卫星细胞衍生的原代成肌细胞生长和分化的影响与底物无关，且比其对C2C12细胞的影响更为显著，这表明弹性蛋白酶在体内对肌生成具有有害作用。此外，弹性蛋白酶损害原代和C2C12成肌细胞向肌管的分化。我们的研究结果证明了中性粒细胞介导的炎症在肌营养不良症中的重要性，并表明弹性蛋白酶介导的成肌细胞行为调节是营养不良肌肉再生能力丧失的潜在机制。
In Duchenne muscular dystrophy, progressive loss of muscle tissue is accompanied by fibrosis, chronic inflammation and reduced muscle regenerative capacity. Although much is known about the development of fibrosis and chronic inflammation in muscular dystrophy, less is known about how they are mechanistically linked to loss of muscle regenerative capacity. We have developed a proteomics method to discover dystrophy-associated changes in the muscle progenitor cell niche, which identified serine proteases, and especially neutrophil elastase, as candidates. We show that elastase activity is increased in dystrophic (mdx4cv) muscle and impairs myoblast survival in culture. While the effect of elastase on C2C12 cell survival correlates with the kinetics of elastase-mediated degradation of the substrate to which the cells adhere, the effect of elastase on satellite cell-derived primary myoblast growth and differentiation is substrate-independent and even more dramatic than the effect on C2C12 cells, suggesting a detrimental role for elastase on myogenesis in vivo. Additionally, elastase impairs differentiation of both primary and C2C12 myoblasts into myotubes. Our findings evidence the importance of neutrophil-mediated inflammation in muscular dystrophy and indicate elastase-mediated regulation of myoblast behaviour as a potential mechanism underlying loss of regenerative capacity in dystrophic muscle.