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人骨骼肌中钠钾ATP酶α和β亚型的单纤维表达及纤维对短期高强度运动训练的特异性适应性

Single-fiber expression and fiber-specific adaptability to short-term intense exercise training of Na+-K+-ATPase α- and β-isoforms in human skeletal muscle.

作者信息

Wyckelsma V L, McKenna M J, Serpiello F R, Lamboley C R, Aughey R J, Stepto N K, Bishop D J, Murphy R M

机构信息

Institute of Sport, Exercise and Active Living (ISEAL), Victoria University, Melbourne, Victoria, Australia; and.

Institute of Sport, Exercise and Active Living (ISEAL), Victoria University, Melbourne, Victoria, Australia; and

出版信息

J Appl Physiol (1985). 2015 Mar 15;118(6):699-706. doi: 10.1152/japplphysiol.00419.2014. Epub 2015 Jan 22.

Abstract

The Na(+)-K(+)-ATPase (NKA) plays a key role in muscle excitability, but little is known in human skeletal muscle about fiber-type-specific differences in NKA isoform expression or adaptability. A vastus lateralis muscle biopsy was taken in 17 healthy young adults to contrast NKA isoform protein relative abundance between type I and IIa fibers. We further investigated muscle fiber-type-specific NKA adaptability in eight of these adults following 4-wk repeated-sprint exercise (RSE) training, comprising three sets of 5 × 4-s sprints, 3 days/wk. Single fibers were separated, and myosin heavy chain (I and IIa) and NKA (α1-3 and β1-3) isoform abundance were determined via Western blotting. All six NKA isoforms were expressed in both type I and IIa fibers. No differences between fiber types were found for α1-, α2-, α3-, β1-, or β3-isoform abundances. The NKA β2-isoform was 27% more abundant in type IIa than type I fibers (P < 0.05), with no other fiber-type-specific trends evident. RSE training increased β1 in type IIa fibers (pretraining 0.70 ± 0.25, posttraining 0.84 ± 0.24 arbitrary units, 42%, P < 0.05). No training effects were found for other NKA isoforms. Thus human skeletal muscle expresses all six NKA isoforms and not in a fiber-type-specific manner; this points to their different functional roles in skeletal muscle cells. Detection of elevated NKA β1 after RSE training demonstrates the sensitivity of the single-fiber Western blotting technique for fiber-type-specific intervention effects.

摘要

钠钾ATP酶(NKA)在肌肉兴奋性中起关键作用,但对于人类骨骼肌中NKA亚型表达或适应性的纤维类型特异性差异知之甚少。对17名健康年轻成年人进行了股外侧肌活检,以对比I型和IIa型纤维之间NKA亚型蛋白的相对丰度。我们进一步研究了其中8名成年人在进行为期4周的重复冲刺运动(RSE)训练后的肌肉纤维类型特异性NKA适应性,该训练包括每周3天,每组进行3组5×4秒的冲刺。分离出单根纤维,并通过蛋白质免疫印迹法测定肌球蛋白重链(I和IIa)以及NKA(α1 - 3和β1 - 3)亚型的丰度。所有六种NKA亚型均在I型和IIa型纤维中表达。在α1、α2、α3、β1或β3亚型丰度方面,未发现纤维类型之间存在差异。NKA β2亚型在IIa型纤维中的丰度比I型纤维高27%(P < 0.05),未发现其他明显的纤维类型特异性趋势。RSE训练使IIa型纤维中的β1增加(训练前0.70±0.25,训练后0.84±0.24任意单位,增加42%,P < 0.05)。未发现其他NKA亚型有训练效果。因此,人类骨骼肌表达所有六种NKA亚型,且并非以纤维类型特异性方式表达;这表明它们在骨骼肌细胞中具有不同的功能作用。RSE训练后检测到NKA β1升高,证明了单纤维蛋白质免疫印迹技术对纤维类型特异性干预效果的敏感性。

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