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急性运动和长期运动对马属动物肌肉中总钠钾ATP酶含量及钠钾ATP酶同工型表达谱的影响。

Effects of acute exercise and long-term exercise on total Na+,K+ -ATPase content and Na+,K+ -ATPase isoform expression profile in equine muscle.

作者信息

van den Burg Maarten M M, Eizema Karin, de Graaf-Roelfsema Ellen, van Breda Eric, Wijnberg Inge D, van der Kolk Johannes H, Everts Maria E

机构信息

Department of Pathobiology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, NL-3584 CL Utrecht, The Netherlands.

出版信息

Am J Vet Res. 2009 Jul;70(7):895-901. doi: 10.2460/ajvr.70.7.895.

Abstract

OBJECTIVE

To investigate the effects of acute exercise and long-term training on Na(+),K(+)-ATPase content, mRNA isoforms, and protein concentration in equine muscle.

ANIMALS

6 Standardbreds.

PROCEDURES

Horses performed a bout of exercise on a treadmill before and after 18 weeks of combined interval and endurance training. Muscle biopsy specimens were obtained from vastus lateralis muscle (VLM) and pectoralis descendens muscle (PDM) before and after exercise. The Na(+),K(+)-ATPase content, mRNA isoforms, and protein concentrations were determined by use of [(3)H]ouabain binding, real-time PCR assay, and western blotting, respectively.

RESULTS

6 Na(+),K(+)-ATPase mRNA isoforms were present in equine muscle, but only A2 and B1 proteins were detected. Exercise before training resulted in increases of mRNA isoforms A1, A2, A3, and B2 in VLM and A1 and B3 in PDM. Training increased resting values for mRNA isoforms A3 and B1 in VLM and B3 in PDM. The Na(+),K(+)-ATPase, [(3)H]ouabain binding, and proteins of mRNA A2 and B1 increased in VLM, whereas in PDM, only A2 protein increased as a result of training. After training, effects of strenuous exercise on mRNA expression were no longer detectable.

CONCLUSIONS AND CLINICAL RELEVANCE

Equine muscle contained all Na(+),K(+)-ATPase mRNA isoforms, but only A2 and B1 proteins could be detected. Expression of these isoforms changed as a result of strenuous exercise and long-term training, representing an adaptive response. Determination of Na(+),K(+)-ATPase gene expression may be relevant for understanding alterations in excitability during neuromuscular diseases.

摘要

目的

研究急性运动和长期训练对马肌肉中钠钾ATP酶含量、mRNA亚型及蛋白质浓度的影响。

动物

6匹标准赛马。

实验步骤

在进行18周的间歇和耐力联合训练前后,让马在跑步机上进行一轮运动。在运动前后从股外侧肌(VLM)和胸降肌(PDM)获取肌肉活检样本。分别使用[³H]哇巴因结合法、实时PCR检测法和蛋白质印迹法测定钠钾ATP酶含量、mRNA亚型及蛋白质浓度。

结果

马肌肉中存在6种钠钾ATP酶mRNA亚型,但仅检测到A2和B1蛋白。训练前运动导致VLM中mRNA亚型A1、A2、A3和B2以及PDM中A1和B3增加。训练增加了VLM中mRNA亚型A3和B1以及PDM中B3的静息值。VLM中钠钾ATP酶、[³H]哇巴因结合以及mRNA A2和B1的蛋白质增加,而在PDM中,训练仅使A2蛋白增加。训练后,剧烈运动对mRNA表达的影响不再可检测到。

结论及临床意义

马肌肉中含有所有钠钾ATP酶mRNA亚型,但仅能检测到A2和B1蛋白。这些亚型的表达因剧烈运动和长期训练而改变,代表一种适应性反应。测定钠钾ATP酶基因表达可能有助于理解神经肌肉疾病期间兴奋性的改变。

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