Myostatin inhibits proliferation of human urethral rhabdosphincter satellite cells
Myostatin inhibits proliferation of human urethral rhabdosphincter satellite cells
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DOI：
10.1111/j.1442-2042.2012.03186.x
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发表时间：
2013-05-01
影响因子：
2.6
通讯作者：
Mimata, Hiromitsu
中科院分区：
文献类型：
--
作者：
Akita, Yasuyuki;Sumino, Yasuhiro;Mimata, Hiromitsu
目的肌生成抑制素（Myostatin）是转化生长因子（TGF）超家族成员之一，是骨骼肌肌细胞发生的负性调节因子。我们研究了肌生长抑制素和肌生长抑制素抑制的拮抗剂，卵泡抑素，对人尿道横纹括约肌卫星细胞（肌肉干细胞）的生长的影响，以开发一种新的策略治疗压力性尿失禁。方法体外培养横纹括约肌卫星细胞，用抗神经细胞粘附分子抗体进行磁性亲和细胞分选。这些细胞用猿猴病毒-40抗原转染以延长它们的寿命。进行细胞增殖试验、细胞周期分析和信号转导研究。内源性肌肉生长抑制素的自分泌作用，通过蛋白质印迹，实时逆转录聚合酶链反应和免疫中和使用抗肌肉生长抑制素抗体进行了评价。结果选择性培养的细胞表达横纹肌标志物，并成功分化为肌管。Myostatin通过Smad 2磷酸化和细胞周期阻滞抑制这些细胞的增殖。肌生长抑制素的抑制作用被卵泡抑素逆转。然而，横纹括约肌卫星细胞似乎没有使用肌生长抑制素的自分泌来调节其增殖。结论抑制肌生长抑制素功能可能是开发刺激横纹括约肌细胞再生治疗压力性尿失禁的新策略的一个有用途径。
Objectives Myostatin, a member of the transforming growth factor- superfamily, is a negative regulator of myogenesis in skeletal muscle. We examined the effect of myostatin and myostatin inhibition by an antagonistic agent, follistatin, on growth of human urethral rhabdosphincter satellite cells (muscle stem cells) to develop a new strategy for treatment of stress urinary incontinence. Methods Rhabdosphincter satellite cells were cultured and selected by magnetic affinity cell sorting using an anti-neural cell adhesion molecule antibody. The cells were transfected with simian virus-40 antigen to extend their lifespan. A cell proliferation assay, a cell cycle analysis and an investigation of signal transduction were carried out. The autocrine action of endogenous myostatin by western blotting, real-time reverse transcription polymerase chain reaction and immunoneutralization using an anti-myostatin antibody was also evaluated. Results Selectively cultured cells expressed markers of striated muscles and successfully differentiated into myotubes. Myostatin inhibited proliferation of these cells through Smad2 phosphorylation and cell cycle arrest. Inhibitory effects of myostatin were reversed by addition of follistatin. However, rhabdosphincter satellite cells did not appear to use autocrine secretion of myostatin to regulate their proliferation. Conclusions Inhibition of myostatin function might be a useful pathway in the development of novel strategies for stimulating rhabdosphincter cells regeneration to treat stress urinary incontinence.