Digoxin and exercise effects on skeletal muscle Na+,K+-ATPase isoform gene expression in healthy humans.

Abstract

In muscle, digoxin inhibits Na+,K+-ATPase (NKA) whereas acute exercise can increase NKA gene expression, consistent with training-induced increased NKA content. We investigated whether oral digoxin increased NKA isoform mRNA expression (qPCR) in muscle at rest, during and post-exercise in 10 healthy adults, who received digoxin (DIG, 0.25mg per day) or placebo (CON) for 14 days, in a randomised, double-blind and cross-over design. Muscle was biopsied at rest, after cycling 20min (10min each at 33%, then 67% ), then to fatigue at 90% and 3h post-exercise. No differences were found between DIG and CON for NKA α1-3 or β1-3 isoform mRNA. Both α1 (354%, P=0.001) and β3 mRNA (P=0.008) were increased 3h post-exercise, with α2 and β1-2 mRNA unchanged, whilst α3 mRNA declined at fatigue (-43%, P=0.045). In resting muscle, total β mRNA (∑(β1+β2+β3)) increased in DIG (60%, P=0.025) and also when transcripts for each isoform were normalised to CON then either summed (P=0.030) or pooled (n=30, P=0.034). In contrast, total α mRNA (∑(α1+α2+α3), P=0.348), normalised then summed (P=0.332), or pooled transcripts (n=30, P=0.717) did not differ with DIG. At rest, NKA α1-2 and β1-2 protein abundances were unchanged by DIG. Post-exercise, α1 and β1-2 proteins were unchanged, but α2 declined at 3h (19%, P=0.020). In conclusion, digoxin did not modify gene expression of individual NKA isoforms at rest or with exercise, indicating NKA gene expression was maintained consistent with protein abundances. However, elevated resting muscle total β mRNA with digoxin suggests a possible underlying β gene-stimulatory effect. HIGHLIGHTS: What is the central question of this study? Na+,K+-ATPase (NKA) in muscle is important for Na+/K+ homeostasis. We investigated whether the NKA-inhibitor digoxin stimulates increased NKA gene expression in muscle and exacerbates NKA gene responses to exercise in healthy adults. What is the main finding and its importance? Digoxin did not modify exercise effects on muscle NKA α1-3 and β1-3 gene transcripts, which comprised increased post-exercise α1 and β3 mRNA and reduced α3 mRNA during exercise. However, in resting muscle, digoxin increased NKA total β isoform mRNA expression. Despite inhibitory-digoxin or acute exercise stressors, NKA gene regulation in muscle is consistent with the maintenance of NKA protein contents.