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酸性成纤维细胞生长因子在体外促进大鼠卫星细胞源性肌管成熟的证据。

Evidence that acidic fibroblast growth factor promotes maturation of rat satellite-cell-derived myotubes in vitro.

作者信息

Düsterhöft S, Pette D

机构信息

Faculty of Biology, University of Konstanz, Germany.

出版信息

Differentiation. 1999 Nov;65(3):161-9. doi: 10.1046/j.1432-0436.1999.6530161.x.

Abstract

Satellite cells isolated from fast tibialis anterior (TA) and slow soleus (SOL) rat muscles were cultivated on matrigel, and treated with acidic fibroblast growth factor (aFGF). The following observations were made: 1) aFGF-treated cultures exhibited enhanced proliferation as mirrored by a twofold increase in DNA content. 2) Compared to the untreated cultures, myotubes in the aFGF cultures were larger; 3) Using reverse transcriptase polymerase chain reaction (RT-PCR) and northern blot analyses, we observed enhanced expression of all adult myosin heavy chain (MHC) isoforms, as well as of myogenin. These findings indicate that, under the culture conditions used, aFGF has a stimulatory effect on proliferation but also on maturation and differentiation of satellite cells. Furthermore, transcript levels of FGF receptor 1 (FGFR1) and 4 (FGFR4) isoforms, as well as of aFGF and bFGF were assessed by RT-PCR. aFGF-treated myotubes displayed increased expression of aFGF and bFGF, suggesting a paracrine effect of exogenous aFGF. In this regard, SOL-derived cultures responded more strongly than TA-derived cultures. The effects of aFGF treatment on the two receptors consisted of a decrease in FGFR1 and an increase in FGFR4 mRNA levels in 5-day-old cultures. In 8-day-old TA cultures, effects of FGF were similar to those in 5-day-old cultures. 8-day FGF-treated SOL cultures treated with FGF for 8 days exhibited higher FGFR1 and FGFR4 mRNA levels than the respective untreated cultures. Compared to 5 day-treated cultures, FGFR1 increased and FGFR4 decreased. This led to a shift in the ratio of FGFR1 to FGFR4 in the FGF-treated cultures which may explain the ability of satellite cells to differentiate under the influence of aFGF.

摘要

从大鼠快速收缩的胫骨前肌(TA)和慢速收缩的比目鱼肌(SOL)中分离出卫星细胞,接种于基质胶上,并给予酸性成纤维细胞生长因子(aFGF)处理。得到以下观察结果:1)aFGF处理的培养物增殖增强,DNA含量增加了两倍。2)与未处理的培养物相比,aFGF培养物中的肌管更大;3)使用逆转录聚合酶链反应(RT-PCR)和Northern印迹分析,我们观察到所有成人肌球蛋白重链(MHC)亚型以及肌细胞生成素的表达增强。这些发现表明,在所使用的培养条件下,aFGF对卫星细胞的增殖、成熟和分化均具有刺激作用。此外,通过RT-PCR评估了FGF受体1(FGFR1)和4(FGFR4)亚型以及aFGF和bFGF的转录水平。aFGF处理的肌管显示aFGF和bFGF的表达增加,表明外源性aFGF具有旁分泌作用。在这方面,源自SOL的培养物比源自TA的培养物反应更强烈。aFGF处理对两种受体的影响包括5日龄培养物中FGFR1水平降低和FGFR4 mRNA水平升高。在8日龄的TA培养物中,FGF的作用与5日龄培养物中的相似。用FGF处理8天的8日龄SOL培养物显示,与各自未处理的培养物相比,FGFR1和FGFR4 mRNA水平更高。与5日龄处理的培养物相比,FGFR1增加而FGFR4减少。这导致FGF处理的培养物中FGFR1与FGFR4的比例发生变化,这可能解释了卫星细胞在aFGF影响下的分化能力。

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