Late-life enalapril administration induces nitric oxide-dependent and independent metabolic adaptations in the rat skeletal muscle.
Recently, we showed that administration of the angiotensin-converting enzyme inhibitor enalapril to aged rats attenuated muscle strength decline and mitigated apoptosis in the gastrocnemius muscle. The aim of the present study was to investigate possible mechanisms underlying the muscle-protective effects of enalapril. We also sought to discern the effects of enalapril mediated by nitric oxide (NO) from those independent of this signaling molecule. Eighty-seven male Fischer 344 × Brown Norway rats were randomly assigned to receive enalapril (n = 23), the NO synthase (NOS) inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME; n = 22), enalapril + L-NAME (n = 19), or placebo (n = 23) from 24 to 27 months of age. Experiments were performed on the tibialis anterior muscle. Total NOS activity and the expression of neuronal, endothelial, and inducible NOS isoforms (nNOS, eNOS, and iNOS) were determined to investigate the effects of enalapril on NO signaling. Transcript levels of tumor necrosis factor-alpha (TNF-α) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) were assessed to explore actions of enalapril on inflammation and mitochondrial biogenesis, respectively. Protein expression of energy-sensing and insulin signaling mediators, including protein kinase B (Akt-1), phosphorylated Akt-1 (pAkt-1), mammalian target of rapamycin (mTOR), AMP-activated protein kinase subunit alpha (AMPKα), phosphorylated AMPKα (pAMPKα), and the glucose transporter GLUT-4, was also determined. Finally, the generation of hydrogen peroxide (H2O2) was quantified in subsarcolemmal (SSM) and intermyofibrillar (IFM) mitochondria. Enalapril increased total NOS activity, which was prevented by L-NAME co-administration. eNOS protein content was enhanced by enalapril, but not by enalapril + L-NAME. Gene expression of iNOS was down-regulated by enalapril either alone or in combination with L-NAME. In contrast, protein levels of nNOS were unaltered by treatments. The mRNA abundance of TNF-α was reduced by enalapril relative to placebo, with no differences among any other group. PCG-1α gene expression was unaffected by enalapril and lowered by enalapril + L-NAME. No differences in protein expression of Akt-1, pAkt-1, AMPKα, pAMPKα, or GLUT-4 were detected among groups. However, mTOR protein levels were increased by enalapril compared with placebo. Finally, all treatment groups displayed reduced SSM, but not IFM H2O2 production relative to placebo. Our data indicate that enalapril induces a number of metabolic adaptations in aged skeletal muscle. These effects result from the concerted modulation of NO and angiotensin II signaling, rather than from a dichotomous action of enalapril on the two pathways. Muscle protection by enalapril administered late in life appears to be primarily mediated by mitigation of oxidative stress and pro-inflammatory signaling.
Department of Aging and Geriatric Research, Institute on Aging, University of Florida, PO Box 100143, Gainesville, FL 32610-0143, USA.
Emanuele
E
Marzetti
Riccardo
R
Calvani
Jameson
J
DuPree
Hazel A
HA
Lees
Silvia
S
Giovannini
Dong-oh
DO
Seo
Thomas W
TW
Buford
Kindal
K
Sweet
Drake
D
Morgan
Kevin Y E
KY
Strehler
Debra
D
Diz
Stephen E
SE
Borst
Natasha
N
Moningka
Karina
K
Krotova
Christy S
CS
Carter
D000222:Adaptation, Physiological / Q000187:drug effects*; D000375:Aging / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000008:administration & dosage; D000818:Animals; D017209:Apoptosis; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004656:Enalapril / Q000008:administration & dosage*; D004734:Energy Metabolism / Q000187:drug effects*; D008297:Male; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D019331:NG-Nitroarginine Methyl Ester / Q000008:administration & dosage; D009569:Nitric Oxide / Q000378:metabolism*; D018384:Oxidative Stress / Q000502:physiology*; D051381:Rats; D011914:Rats, Inbred BN; D011916:Rats, Inbred F344
D000806:Angiotensin-Converting Enzyme Inhibitors; D009569:Nitric Oxide; D004656:Enalapril; D019331:NG-Nitroarginine Methyl Ester