Investigation of gene expression in C(2)C(12) myotubes following simvastatin application and mechanical strain.
OBJECTIVE
The 3-hydroxy-3methylgutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) are the most effective prescribed drugs for lowering serum cholesterol; however, although statins are extremely safe medications and have brought significant benefits to patients with hypercholesterolemia, they have been shown to produce myalgia, cramps, exercise intolerance and fatigue. The aim of the study was to investigate the molecular mechanisms that may mediate statin myopathy.


METHODS
We used DNA microarray analysis to examine the changes in gene expression profiles induced by 1 hour and 6 hours of statin treatment on differentiated C(2)C(12) myotubes. Four genes were selected for analysis at the protein level using Western blot analysis on myotubes treated with statin with or without additional mechanical stretching.


RESULTS
Eighty-five genes exhibited more than a 2-fold up- or down-regulation in expression, of which 46 have known biological functions related primarily to transmembrane transport, signal transduction, cell growth/maintenance, protein metabolism, or apoptosis. At protein level, three of the four proteins were induced (Adrb1, Socs4 and Cflar) and one was repressed (Birc4). Changes in protein expression largely mirrored the changes in their corresponding transcripts, although the fold-change was less dramatic. The addition of imposed muscle fiber stretching did not exacerbate the expression of these genes at the protein level with the exception of Cflar, a pro-apoptotic protein.


CONCLUSIONS
These data suggested that alterations in the expressions of some statin-regulated genes could be causative factors for statin toxicity in muscle. Repression of the anti-apoptosis gene (Birc4) and activation of the pro-apoptosis gene (Cflar) indicated that cell death may play an important role in statin-induced myopathy.
Yu
Ji-Guo
JG
Department of Surgical and Perioperative Science, Sports Medicine Unite, Umeå University, Umeå, Sweden. jiguo.yu@idrott.umu.se
Sewright
Kimberly
K
Hubal
Monica J
MJ
Liu
Jing-Xia
JX
Schwartz
Lawrence M
LM
Hoffman
Eric P
EP
Clarkson
Priscilla M
PM
D000818:Animals; D053446:CASP8 and FADD-Like Apoptosis Regulating Protein / Q000235:genetics; D002478:Cells, Cultured; D015870:Gene Expression; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000494:pharmacology*; D051379:Mice; D046228:Microarray Analysis; D018485:Muscle Fibers, Skeletal / Q000187:drug effects* / Q000502:physiology; D019821:Simvastatin / Q000494:pharmacology*; D013314:Stress, Mechanical; D051636:X-Linked Inhibitor of Apoptosis Protein / Q000235:genetics
D053446:CASP8 and FADD-Like Apoptosis Regulating Protein; C506386:CFLAR protein, human; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D051636:X-Linked Inhibitor of Apoptosis Protein; C496866:XIAP protein, human; D019821:Simvastatin