Osteopontin expression in coculture of differentiating rat fetal skeletal fibroblasts and myoblasts
Osteopontin expression in coculture of differentiating rat fetal skeletal fibroblasts and myoblasts
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分化大鼠胎儿骨骼成纤维细胞和成肌细胞共培养中骨桥蛋白的表达
DOI：
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
L. Carvalho
中科院分区：
文献类型：
--
作者：
R. Pereira;S. Carvalho;A. Stumbo;C. A. Rodrigues;L. C. Porto;A. Moura;L. Carvalho
体外培养的皮肤成纤维细胞可通过生物化学和形态学特征的发育获得肌成纤维细胞表型，主要是α-平滑肌肌动蛋白（α-SMA）的表达。成肌分化是骨骼肌发育的中心事件，并且通常在骨骼肌发育和再生的背景下在体外进行研究。控制这一过程的是成肌细胞和细胞外基质之间的一系列复杂的相互作用。骨桥蛋白（OPN）是一种酸性磷酸化基质蛋白，含有Arg-Gly-Asp（RGD）细胞附着序列，已被鉴定为几种细胞类型的粘附和迁移底物。本研究的目的是探讨骨桥蛋白的表达过程中的骨骼成纤维细胞分化成肌成纤维细胞和在肌发生在共培养模型。采用酶解法分离18日龄Wistar系大鼠胚胎骨骼肌成纤维细胞和成肌细胞。在第1和9天，免疫标记共培养物，并分别对细胞进行Western印迹分析OPN表达。我们的数据使用共聚焦显微镜显示，成肌细胞显示出强烈的OPN染色，这种标记后肌管分化保持。相反，在成纤维细胞分化为肌成纤维细胞的过程中，我们观察到OPN表达显著增加。通过免疫标记获得的结果通过Western印迹法证实。我们认为，OPN是重要的，主要是在早期阶段的肌发生，促进成肌细胞的融合和分化，并在肌成纤维细胞中的OPN的表达增加可能与其作为一个关键的细胞因子调节组织修复和炎症的影响。
SummarySkeletal fibroblasts in vitro can acquire myofibroblast phenotypes by the development of biochemical and morphological features, mainly the expression of alpha-smooth-muscle actin (α-SMA). Myogenic differentiation is a central event in skeletal muscle development, and has commonly been studied in vitro in the context of skeletal muscle development and regeneration. Controlling this process is a complex set of interactions between myoblasts and the extracellular matrix. Osteopontin (OPN) is an acidic, phosphorylated matrix protein that contains an Arg-Gly-Asp (RGD) cell attachment sequence and has been identified as an adhesive and migratory substrate for several cell types. The aim of this study was to investigate osteopontin expression during the differentiation of skeletal fibroblasts into myofibroblasts and during myogenesis in a coculture model. Fibroblasts and myoblasts were obtained from skeletal muscle of 18-d-old Wistar strain rat fetuses by enzymatic dissociation. At 1 and 9 d, cocultures were immunolabeled, and the cells were also separately subjected to Western blotting to analyze OPN expression. Our data using confocal microscopy showed that myoblasts displayed a strong staining for OPN and that this labeling was maintained after myotube differentiation. Conversely, during fibroblast differentiation into myofibroblasts, we observed a significant increase in OPN expression. The results obtained by immunolabeling were confirmed by Western blotting. We suggest that OPN is important mainly during early stages of myogenesis, facilitating myoblast fusion and differentiation, and that the increased expression of OPN in myofibroblasts might be related to its effects as a key cytokine regulating tissue repair and inflammation.
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影响因子：
8.3
作者：
Singh, K;Sirokman, G;Colucci, WS
通讯作者：
Colucci, WS
DOI：
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发表时间：
1994-12
期刊：
The American journal of pathology
影响因子：
--
作者：
C. Murry;C. Giachelli;S. Schwartz;R. Vracko
通讯作者：
C. Murry;C. Giachelli;S. Schwartz;R. Vracko
影响因子：
37.8
作者：
Graf, K;Do, YS;Hsueh, WA
通讯作者：
Hsueh, WA
影响因子：
1.6
作者：
Bayless, KJ;Davis, GE;Meininger, GA
通讯作者：
Meininger, GA
影响因子：
20.1
作者：
LIAW, L;ALMEIDA, M;GIACHELLI, CM
通讯作者：
GIACHELLI, CM