Peroxisome proliferator-activated receptor alpha agonists enhance cardiomyogenesis of mouse ES cells by utilization of a reactive oxygen species-dependent mechanism.
Peroxisome proliferator-activated receptors (PPARalpha, -beta and -gamma) are nuclear receptors involved in transcriptional regulation of lipid and energy metabolism. Since the energy demand increases when cardiac progenitor cells are developing rhythmic contractile activity, PPAR activation may play a critical role during cardiomyogenesis of embryonic stem (ES) cells. It is shown that ES cells express PPARalpha, -beta, and -gamma mRNA during differentiation of ES cells towards cardiac cells. Treatment with PPARalpha agonists (WY14643, GW7647, and ciprofibrate) significantly increased cardiomyogenesis and expression of the cardiac genes MLC2a, ANP, MHC-beta, MLC2v, and cardiac alpha-actin. Furthermore, WY14643 increased PPARalpha gene expression and the expression of the cardiogenic transcription factors GATA-4, Nkx2.5, DTEF-1, and MEF 2C. In contrast, the PPARalpha antagonist MK886 decreased cardiomyogenesis, whereas the PPARbeta agonist L-165,041 as well as the PPARgamma agonist GW1929 were without effects. Treatment with PPARalpha, but not PPARbeta, and PPARgamma agonists and MK886, resulted in generation of reactive oxygen species (ROS), which was inhibited in the presence of the NADPH oxidase inhibitors diphenylen iodonium (DPI) and apocynin and the free radical scavengers vitamin E and N-(2-mercapto-propionyl)-glycine (NMPG), whereas the mitochondrial complex I inhibitor rotenone was without effects. The effect of PPARalpha agonists on cardiomyogenesis of ES cells was abolished upon preincubation with free radical scavengers and NADPH oxidase inhibitors, indicating involvement of ROS in PPARalpha, mediated cardiac differentiation. In summary, our data indicate that stimulation of PPARalpha but not PPARbeta and -gamma enhances cardiomyogenesis in ES cells using a pathway that involves ROS and NADPH oxidase activity.
Sharifpanah
Fatemeh
F
Department of Physiology, Justus-Liebig-University Giessen, Aulweg 129, 35392 Giessen, Germany.
Wartenberg
Maria
M
Hannig
Madeleine
M
Piper
Hans-Michael
HM
Sauer
Heinrich
H
D000818:Animals; D002087:Butyrates / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D002995:Clofibric Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D058607:Fibric Acids; D016166:Free Radical Scavengers / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006321:Heart / Q000196:embryology; D007150:Immunohistochemistry; D007211:Indoles / Q000494:pharmacology; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D019255:NADPH Oxidases / Q000187:drug effects / Q000378:metabolism; D047493:PPAR alpha / Q000819:agonists*; D047495:PPAR gamma / Q000187:drug effects / Q000378:metabolism; D048068:PPAR-beta / Q000187:drug effects / Q000378:metabolism; D020025:Peroxisome Proliferators / Q000494:pharmacology; D010671:Phenylurea Compounds / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D017382:Reactive Oxygen Species / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D002722:Chlorobenzenes
D002087:Butyrates; D002995:Clofibric Acid; D004791:Enzyme Inhibitors; D058607:Fibric Acids; D016166:Free Radical Scavengers; D007211:Indoles; D019255:NADPH Oxidases; D047493:PPAR alpha; D047495:PPAR gamma; D048068:PPAR-beta; D020025:Peroxisome Proliferators; D010671:Phenylurea Compounds; D011743:Pyrimidines; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; C453899:GW 7647; C060893:MK-886; D002722:Chlorobenzenes; C006253:pirinixic acid; C019304:ciprofibrate