Department of Movement Sciences and Wellness, University Parthenope, Naples, Italy.
CEINGE-Biotecnologie Avanzate, Naples, Italy.
Eur J Appl Physiol. 2021 Oct;121(10):2903-2912. doi: 10.1007/s00421-021-04733-1. Epub 2021 Jul 1.
Regular exercise affects the expression of several genes, proteins and microRNAs (miRNAs) in time- and intensity-dependent manner promoting longevity. We previously identified from GeneChip Array analysis several differentially expressed genes and miRNAs in muscle from veteran football players (VPG) compared to active untrained elderly subjects (CG); here we focussed on miRNA-1303 (miR-1303). The aims of the present research were: to analyse the effects of football training on the expression of miR-1303 and to identify its putative target involved in the longevity pathways in skeletal muscle from VPG compared to CG.
RNA samples from 12 VPG and 12 CG muscle biopsies were used to validate miR-1303 expression. Crossing four different bioinformatic algorithms, we identified 16 putative targets of miR-1303; from these, BAG-2, KLHL7 and KBTBD6 were chosen for further validation by Western blot analysis in LHCN-M2 human myoblasts transiently transfected with miR-1303.
Football training down-regulates miR-1303 expression in muscle from VPG compared to CG and the expression of BAG-2, a chaperon protein involved in the autophagy pathway, inversely correlated to overexpression of miR-1303 in a time-dependent manner, indicating that it is a miR-1303 potential target.
This is the first report, to our knowledge, describing miR-1303 regulation in skeletal muscle by football training and the identification of a target protein, BAG-2, involved in the autophagy pathway. This result contributes to the enlargement of knowledge on the molecular mechanisms linking football training, autophagy and longevity.
有规律的运动以时间和强度依赖的方式影响着多个基因、蛋白质和 microRNAs(miRNAs)的表达,从而促进长寿。我们之前通过基因芯片分析,从资深足球运动员(VPG)的肌肉中鉴定出了一些与活跃未经训练的老年人(CG)相比差异表达的基因和 miRNAs;在这里我们重点关注 microRNA-1303(miR-1303)。本研究的目的是:分析足球训练对 miR-1303 表达的影响,并在 VPG 与 CG 的骨骼肌中,鉴定其参与长寿途径的潜在靶基因。
使用 12 名 VPG 和 12 名 CG 的肌肉活检 RNA 样本验证 miR-1303 的表达。通过交叉四种不同的生物信息学算法,我们鉴定出 16 个 miR-1303 的潜在靶基因;从这些基因中,选择 BAG-2、KLHL7 和 KBTBD6 用于通过瞬时转染 miR-1303 的 LHCN-M2 人成肌细胞进行 Western blot 分析进一步验证。
与 CG 相比,足球训练下调了 VPG 肌肉中 miR-1303 的表达,而自噬途径中参与伴侣蛋白 BAG-2 的表达与 miR-1303 的过表达呈时间依赖性负相关,表明其是 miR-1303 的潜在靶基因。
这是第一个报道足球训练调节骨骼肌中 miR-1303 表达的报告,并且鉴定出了自噬途径中的靶蛋白 BAG-2。这一结果丰富了足球训练、自噬和长寿之间分子机制的知识。