Cyclosporin A treatment upregulates Id1 and Smad3 expression and delays skeletal muscle regeneration
Cyclosporin A treatment upregulates Id1 and Smad3 expression and delays skeletal muscle regeneration
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DOI：
10.1007/s00401-005-1049-x
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发表时间：
2005-09-01
影响因子：
12.7
通讯作者：
Yasuhara, M
中科院分区：
文献类型：
--
作者：
Sakuma, K;Nakao, R;Yasuhara, M
与肌肉再生有关的分子信号通路尚未被确定。先前，我们证明了用环孢素A（一种钙调磷酸酶抑制剂）治疗的小鼠在肌肉损伤后不能正常再生。通过反转录(RT)-PCR、Western blot和免疫组化分析，我们研究了CsA治疗后再生肌肉中活化T细胞核因子（NFAT）、肌细胞增强因子2 （MEF2）、MyoD家族、Id-1和Smad3的数量是否发生变化。将成年雄性ICR小鼠胫骨前肌注射布比卡因，并给予CsA （25 mg/kg）或载药，每日1次。分别于受伤后1、2、4、6、9、14天死亡。RT-PCR分析没有显示安慰剂组和csa组小鼠再生肌肉中MEF2s、MyoD和肌原素mRNA水平的显著差异。相比之下，在术后4天和9天，与安慰剂治疗的小鼠相比，服用csa的小鼠MRF4 mRNA显著增加。在csa处理的小鼠中，Id1 mRNA水平在第9天相对于安慰剂处理的小鼠升高。6天后，csa处理小鼠再生肌肉的许多卫星细胞和/或成肌细胞样细胞中具有更丰富的增殖细胞核抗原（PCNA）和cyclin D1蛋白。CsA组小鼠肌肉生长抑制素、tgf - β 2和Smad3 mRNA和蛋白的表达量明显增加。9天后，许多卫星细胞和/或成肌细胞在CsA-治疗小鼠中显示MyoD和Smad3的明显共定位，而在安慰剂-治疗小鼠中则没有。我们的研究结果表明，CsA处理上调Id1和Smad3的表达，延缓骨骼肌再生。
The molecular signaling pathway linked to muscle regeneration has not yet been identified. Previously, we demonstrated that mice treated with cyclosporin A (CsA), a calcineurin inhibitor, failed to regenerate normally after muscle damage. Using reverse transcription (RT)-PCR, Western blot and immunohistochemical analysis, we investigated whether the amounts of nuclear factor of activated T cells (NFAT), myocyte-enhancer factor 2 (MEF2), the MyoD family, Id-1, and Smad3 change in the regenerating muscle after CsA treatment. Adult male ICR mice were subjected to a bupivacaine injection into the tibialis anterior muscle, and were treated with either CsA (25 mg/kg) or vehicle once daily. They were killed at 1, 2, 4, 6, 9 and 14 days post injury. RT-PCR analysis did not show a significant difference in MEF2s, MyoD and myogenin mRNA levels in the regenerating muscle in either placebo- and CsA-administered mice. In contrast, a significant increase in MRF4 mRNA was seen in CsA-administered mice compared to the placebo-treated mice at 4 and 9 days post surgery. In CsA-treated mice, the level of Id1 mRNA was elevated at day 9 relative to the placebo-treated mice. After 6 days, the CsA-treated mice possessed more abundant proliferating cell nuclear antigen (PCNA) and cyclin D1 protein in many satellite cells and/or myoblast-like cells in the regenerating muscle. The amount of myostatin, TGF-beta 2 and Smad3 mRNA and proteins was increased more markedly in the mice treated with CsA. After 9 days, many satellite cells and/or myoblasts showed apparent co-localization of both MyoD and Smad3 in CsA-, but not in placebo-, treated mice. Our results demonstrated that CsA treatment upregulates Id1 and Smad3 expression and delays skeletal muscle regeneration in vivo.