Thomassen Martin, Gunnarsson Thomas P, Christensen Peter M, Pavlovic Davor, Shattock Michael J, Bangsbo Jens
Department of Nutrition, Exercise and Sports, Section of Integrated Physiology, University of Copenhagen, Copenhagen, Denmark; and.
Cardiovascular Division, King's College London, The Rayne Institute, St. Thomas' Hospital, London, United Kingdom.
Am J Physiol Regul Integr Comp Physiol. 2016 Apr 1;310(7):R659-69. doi: 10.1152/ajpregu.00081.2015. Epub 2016 Jan 20.
The present study examined the effect of intensive training in combination with marked reduction in training volume on phospholemman (FXYD1) expression and phosphorylation at rest and during exercise. Eight well-trained cyclists replaced their regular training with speed-endurance training (10-12 × ∼30-s sprints) two or three times per week and aerobic high-intensity training (4-5 × 3-4 min at 90-95% of peak aerobic power output) 1-2 times per week for 7 wk and reduced the training volume by 70%. Muscle biopsies were obtained before and during a repeated high-intensity exercise protocol, and protein expression and phosphorylation were determined by Western blot analysis. Expression of FXYD1 (30%), actin (40%), mammalian target of rapamycin (mTOR) (12%), phospholamban (PLN) (16%), and Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) γ/δ (25%) was higher (P < 0.05) than before the training intervention. In addition, after the intervention, nonspecific FXYD1 phosphorylation was higher (P < 0.05) at rest and during exercise, mainly achieved by an increased FXYD1 Ser-68 phosphorylation, compared with before the intervention. CaMKII, Thr-287, and eukaryotic elongation factor 2 Thr-56 phosphorylation at rest and during exercise, overall PKCα/β, Thr-638/641, and mTOR Ser-2448 phosphorylation during repeated intense exercise as well as resting PLN Thr-17 phosphorylation were also higher (P < 0.05) compared with before the intervention period. Thus, a period of high-intensity training with reduced training volume increases expression and phosphorylation levels of FXYD1, which may affect Na(+)/K(+) pump activity and muscle K(+) homeostasis during intense exercise. Furthermore, higher expression of CaMKII and PLN, as well as increased phosphorylation of CaMKII Thr-287 may have improved intracellular Ca(2+) handling.
本研究考察了强化训练结合训练量显著减少对静息及运动时磷蛋白(FXYD1)表达和磷酸化的影响。八名训练有素的自行车运动员将其常规训练替换为每周两到三次的速度耐力训练(10 - 12次约30秒冲刺)以及每周一到两次的有氧高强度训练(4 - 5次,每次3 - 4分钟,强度为峰值有氧功率输出的90 - 95%),持续7周,并将训练量减少了70%。在重复高强度运动方案之前和期间获取肌肉活检样本,通过蛋白质印迹分析确定蛋白质表达和磷酸化情况。FXYD1(30%)、肌动蛋白(40%)、雷帕霉素哺乳动物靶点(mTOR)(12%)、受磷蛋白(PLN)(16%)和钙/钙调蛋白依赖性蛋白激酶II(CaMKII)γ/δ(25%)的表达高于训练干预前(P < 0.05)。此外,干预后,与干预前相比,静息及运动时非特异性FXYD1磷酸化水平更高(P < 0.05),主要是通过FXYD1丝氨酸68磷酸化增加实现的。静息及运动时CaMKII苏氨酸287和真核生物延伸因子2苏氨酸56的磷酸化、重复高强度运动期间总体蛋白激酶Cα/β苏氨酸638/641以及mTOR丝氨酸2448的磷酸化以及静息时PLN苏氨酸17的磷酸化也高于干预前时期(P < 0.05)。因此,一段训练量减少的高强度训练期会增加FXYD1的表达和磷酸化水平,这可能会在剧烈运动期间影响钠/钾泵活性和肌肉钾稳态。此外,CaMKII和PLN表达的增加以及CaMKII苏氨酸287磷酸化的增加可能改善了细胞内钙处理。