Downregulation of let-7 by Electrical Acupuncture Increases Protein Synthesis in Mice.
Downregulation of let-7 by Electrical Acupuncture Increases Protein Synthesis in Mice.
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DOI：
10.3389/fphys.2021.697139
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发表时间：
2021
影响因子：
4
通讯作者：
Wang XH
中科院分区：
文献类型：
--
作者：
Huang Y;Yu M;Kuma A;Klein JD;Wang Y;Hassounah F;Cai H;Wang XH
我们之前的研究发现，低频电刺激针灸（Acu/LFES）通过减弱小鼠蛋白质降解来防止肌肉萎缩。目前的研究探讨了 Acu/LFES 对蛋白质合成的影响。 C57/BL6小鼠后肢接受Acu/LFES治疗一次，每次30分钟。 Acu/LFES 点根据 WHO 标准针灸命名法选择，并使用 SDZ-II 电子针灸仪施加电刺激。通过表面传感翻译（SUnSET）测定来测量肌肉蛋白质合成。使用连续离心分离外泌体，并使用 NanoSight 仪器测量收集的外泌体的浓度和大小。使用高灵敏度 DNA 芯片验证了血清外泌体中的成熟 microRNA 文库。后肢和前肢肌肉中的蛋白质合成均得到增强。用 GW4869 阻断外泌体分泌可减少 Acu/LFES 诱导的蛋白质合成增加。 MicroRNA 深度测序表明，血清外泌体中 Let-7 miRNA 家族的四个成员显着减少。实时 qPCR 进一步验证了 Acu/LFES 介导的血清外泌体和骨骼肌中 let-7c-5p 的降低。在培养的 C2C12 肌管中，let-7c 的抑制不仅增加了蛋白质合成，而且还增加了 Igf1 和 Igf1 受体的蛋白质丰度。使用荧光素酶报告基因检测，我们证明 let-7 直接抑制 Igf1。后肢上的 Acu/LFES 减少 let-7-5p，导致 Igf1 信号上调并增加后肢和前肢骨骼肌中的蛋白质合成。这为电针治疗如何积极影响肌肉健康提供了新的认识。
Our previous study found that acupuncture with low frequency electrical stimulation (Acu/LFES) prevents muscle atrophy by attenuation of protein degradation in mice. The current study examines the impact of Acu/LFES on protein synthesis. C57/BL6 mice received Acu/LFES treatment on hindlimb for 30 min once. Acu/LFES points were selected by WHO Standard Acupuncture Nomenclature and electric stimulation applied using an SDZ-II Electronic acupuncture instrument. Muscle protein synthesis was measured by the surface-sensing of translation (SUnSET) assay. Exosomes were isolated using serial centrifugation and concentration and size of the collected exosomes were measured using a NanoSight instrument. The mature microRNA library in serum exosomes was validated using a High Sensitivity DNA chip. Protein synthesis was enhanced in the both hindlimb and forelimb muscles. Blocking exosome secretion with GW4869 decreased the Acu/LFES-induced increases in protein synthesis. MicroRNA-deep sequencing demonstrated that four members of the Let-7 miRNA family were significantly decreased in serum exosomes. Real time qPCR further verified Acu/LFES-mediated decreases of let-7c-5p in serum exosomes and skeletal muscles. In cultured C2C12 myotubes, inhibition of let-7c not only increased protein synthesis, but also enhanced protein abundance of Igf1 and Igf1 receptors. Using a luciferase reporter assay, we demonstrated that let-7 directly inhibits Igf1. Acu/LFES on hindlimb decreases let-7-5p leading to upregulation of the Igf1 signaling and increasing protein synthesis in both hindlimb and forelimb skeletal muscles. This provides a new understanding of how the electrical acupuncture treatment can positively influence muscle health.
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影响因子：
29
作者：
Dubinsky AN;Dastidar SG;Hsu CL;Zahra R;Djakovic SN;Duarte S;Esau CC;Spencer B;Ashe TD;Fischer KM;MacKenna DA;Sopher BL;Masliah E;Gaasterland T;Chau BN;Pereira de Almeida L;Morrison BE;La Spada AR
通讯作者：
La Spada AR
影响因子：
1
作者：
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通讯作者：
Liang, Xibin
影响因子：
56.9
作者：
Mayr, Christine;Hemann, Michael T.;Bartel, David P.
通讯作者：
Bartel, David P.
影响因子：
4.8
作者：
Cappelletti, Cristina;Galbardi, Barbara;Mantegazza, Renato
通讯作者：
Mantegazza, Renato
影响因子：
64.8
作者：
Reinhart, BJ;Slack, FJ;Ruvkun, G
通讯作者：
Ruvkun, G