Insulin-like growth factor-1 mediates stretch-induced upregulation of myostatin expression in neonatal rat cardiomyocytes.
OBJECTIVE
Myostatin, a negative regulator of muscle growth, is increased in hypertrophied and infarcted heart. However, the mechanism of regulation is not known. Mechanical stress is an important regulatory factor for cardiomyocyte growth. The aim of the study was to investigate the effect of cyclic stretch on the expression of myostatin gene in cardiomyocytes.


METHODS
Neonatal Wistar rat cardiomyocytes grown on a flexible membrane base were stretched by vacuum to 20% of maximum elongation at 60 cycles/min. An in vivo model of aorta-caval shunt in adult rats was used to investigate the myostatin expression.


RESULTS
Cyclic stretch significantly increased myostatin protein and mRNA expression after 6 to 18 h of stretch. Addition of the p38 mitogen-activated protein (MAP) kinase inhibitor SB203580, insulin-like growth factor-1 (IGF-1) monoclonal antibody, and p38 siRNA 30 min before stretch inhibited the induction of myostatin protein. Cyclic stretch increased, while SB203580, IGF-1, and IGF-1 receptor antibody abolished, the phosphorylated p38 protein. Gel shift assays showed significant increase of DNA-protein binding activity of myocyte enhancer factor 2 (MEF2) after stretch, and transfection with p38 siRNA abolished the DNA-protein binding activity induced by cyclic stretch. Cyclic stretch significantly increased the IGF-1 secretion from myocytes. Both conditioned media from stretched myocytes and exogenous administration of IGF-1 recombinant protein to the non-stretched myocytes increased myostatin protein expression similar to that seen after cyclic stretch. An in vivo model of aorta-caval shunt in adult rats also demonstrated the increased myostatin expression in the myocardium.


CONCLUSIONS
Cyclic mechanical stretch enhances myostatin expression in cultured rat neonatal cardiomyocytes. The stretch-induced myostatin is mediated by IGF-1 at least in part through a p38 MAP kinase and MEF2 pathway.
Shyu
Kou-Gi
KG
Department of Education and Research, Shin Kong Wu Ho-Su Memorial Hospital, 95 Wen-Chang Road, Taipei 111, Taiwan. shyukg@ms12.hinet.net
Ko
Wei-Hsu
WH
Yang
Wei-Shiung
WS
Wang
Bao-Wei
BW
Kuan
Peiliang
P
D000714:Anastomosis, Surgical; D000818:Animals; D000831:Animals, Newborn; D001011:Aorta / Q000601:surgery; D015152:Blotting, Northern / Q000379:methods; D015153:Blotting, Western / Q000379:methods; D002478:Cells, Cultured; D024202:Electrophoretic Mobility Shift Assay; D004789:Enzyme Activation; D015870:Gene Expression; D007334:Insulin-Like Growth Factor I / Q000378:metabolism / Q000494:pharmacology*; D048670:MAP Kinase Kinase 4 / Q000032:analysis / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000032:analysis / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000378:metabolism*; D055435:Myostatin; D051381:Rats; D017208:Rats, Wistar; D013314:Stress, Mechanical; D016212:Transforming Growth Factor beta / Q000032:analysis / Q000378:metabolism*; D015854:Up-Regulation / Q000187:drug effects*; D014684:Venae Cavae / Q000601:surgery; D048051:p38 Mitogen-Activated Protein Kinases / Q000032:analysis / Q000378:metabolism
C525913:Mstn protein, rat; D055435:Myostatin; D016212:Transforming Growth Factor beta; D007334:Insulin-Like Growth Factor I; D019950:Mitogen-Activated Protein Kinase 1; D048051:p38 Mitogen-Activated Protein Kinases; D048670:MAP Kinase Kinase 4