Differences in transcriptional patterns of extracellular matrix, inflammatory, and myogenic regulatory genes in myofibroblasts, fibroblasts, and muscle precursor cells isolated from old male rat skeletal muscle using a novel cell isolation procedure.
Differences in transcriptional patterns of extracellular matrix, inflammatory, and myogenic regulatory genes in myofibroblasts, fibroblasts, and muscle precursor cells isolated from old male rat skeletal muscle using a novel cell isolation procedure.
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使用新型细胞分离程序从老年雄性大鼠骨骼肌中分离出的肌成纤维细胞、成纤维细胞和肌肉前体细胞中细胞外基质、炎症和肌源性调节基因的转录模式差异。
DOI：
10.1007/s10522-012-9382-7
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发表时间：
2012
期刊：
影响因子：
4.5
通讯作者：
Booth,FrankW
中科院分区：
文献类型：
--
作者：
Zwetsloot,KevinA;Nedergaard,Anders;Gilpin,LeighT;Childs,ThomasE;Booth,FrankW
老化的骨骼肌显示纤维化增加和再生受损。虽然还没有很好地表征骨骼肌成纤维细胞如何促成这些现象，但转化生长因子-β1（TGF-β1）和Delta/Notch信号传导已涉及影响肌肉再生。在这项研究中，在分化成纤维细胞（肌成纤维细胞），增殖成纤维细胞和肌肉前体细胞（MPC）中鉴定了衰老表型的独特组合，这些细胞表征了在老年骨骼肌中观察到的受损再生潜力。采用一种新的双重分离技术，即从同一大鼠骨骼肌样品中分离成纤维细胞和MPC，并在5%O2和5%CO2的细胞培养条件下，我们首次报道了32月龄骨骼肌的肌成纤维细胞与3月龄相比，显示出基本细胞外基质（ECM）基因胶原4α1的mRNA水平增加（增加83%）、胶原蛋白4α2（增加98%）和层粘连蛋白2（增加113%），以及炎症标志物白细胞介素-6（4.3倍增加）和肿瘤坏死因子α（3.2倍增加）以及TGF-β1（84%增加）的mRNA水平增加，其蛋白质控制增殖和分化。此外，我们证明了32月龄骨骼肌的增殖成纤维细胞显示Notch配体Delta 1的mRNA水平增加（增加≥2.0倍）。总之，这些研究结果表明，ECM和炎症基因在32月龄骨骼肌肌成纤维细胞中的表达增加可能有助于损害老年骨骼肌再生的纤维化表型。此外，我们相信，新的双重隔离技术，在这里开发的研究，调查骨骼肌中的MPC，成纤维细胞和肌成纤维细胞之间的通信可能是有用的。
Aged skeletal muscle displays increased fibrosis and impaired regeneration. While it is not well characterized how skeletal muscle fibroblasts contribute to these phenomena, transforming growth factor-β1 (TGF-β1) and Delta/Notch signaling have been implicated to influence muscle regeneration. In this study, a unique combination of aging phenotypes is identified in differentiating fibroblasts (myofibroblasts), proliferating fibroblasts, and muscle precursor cells (MPCs) that characterize an impaired regenerative potential observed in aged skeletal muscle. Using a novel dual-isolation technique, that isolates fibroblasts and MPCs from the same rat skeletal muscle sample, and cell culture conditions of 5 % O2and 5 % CO2, we report for the first time that myofibroblasts from 32-mo-old skeletal muscle, compared to 3-mo-old, display increased levels of mRNA for the essential extracellular matrix (ECM) genes, collagen 4α1 (83 % increase), collagen 4α2 (98 % increase), and laminin 2 (113 % increase), as well as increased levels of mRNA for the inflammatory markers, interleukin-6 (4.3-fold increase) and tumor necrosis factor α (3.2-fold increase), and TGF-β1 (84 % increase), whose protein controls proliferation and differentiation. Additionally, we demonstrate that proliferating fibroblasts from 32-mo-old skeletal muscle display increased levels of mRNA for the Notch ligand, Delta 1 (≥2.0-fold increase). Together, these findings suggest that increased expression of ECM and inflammatory genes in myofibroblasts from 32-mo-old skeletal muscle may contribute to the fibrogenic phenotype that impairs regeneration in aged skeletal muscle. Furthermore, we believe the novel dual-isolation technique developed here may be useful in studies that investigate communications among MPCs, fibroblasts, and myofibroblasts in skeletal muscle.