Thromboxane a(2) induces differentiation of human mesenchymal stem cells to smooth muscle-like cells.
Thromboxane A(2) (TxA(2)) is involved in smooth muscle contraction and atherosclerotic vascular diseases. Accumulating evidence suggests a pivotal role for mesenchymal stem cells (MSCs) in vascular remodeling. In the present study, we demonstrate for the first time that the TxA(2) mimetic U46619 induces differentiation of human adipose tissue-derived MSCs (hADSCs) to smooth muscle-like cells, as demonstrated by increased expression of smooth muscle-specific contractile proteins such as alpha-smooth muscle actin (alpha-SMA), calponin, smoothelin, and smooth muscle-myosin heavy chain. Using an in vitro collagen gel lattice contraction assay, we showed that U46619-induced expression of the contractile proteins was associated with increased contractility of the cells. U46619 increased the intracellular Ca(2+) concentration in hADSCs and pretreatment of the cells with the thromboxane receptor antagonist SQ29548 or the calmodulin (CaM) inhibitor W13 abrogated the U46619-induced alpha-SMA expression and contractility, suggesting a pivotal role of Ca(2+)/CaM in the U46619-stimulated smooth muscle differentiation of hADSCs. In addition, U46619 elicited activation of RhoA in hADSCs, and pretreatment of the cells with the Rho kinase-specific inhibitor Y27632 or overexpression of the dominant-negative mutants of RhoA and Rho kinase blocked U46619-stimulated alpha-SMA expression and contractility. Furthermore, U46619 induced phosphorylation of myosin light chain (MLC) through CaM/MLC kinase- and Rho kinase-dependent pathways, and the MLC kinase inhibitor ML-7 abrogated U46619-induced alpha-SMA expression and contractility. These results suggest that U46619 induces differentiation of hADSCs to contractile smooth muscle-like cells through CaM/MLCK- and RhoA-Rho kinase-dependent actin polymerization.
Medical Research Center for Ischemic Tissue Regeneration, the Medical Research Institute, Department of Physiology, Busan, Republic of Korea.
Mi Ra
MR
Kim
Eun Su
ES
Jeon
Young Mi
YM
Kim
Jung Sub
JS
Lee
Jae Ho
JH
Kim
D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid / Q000494:pharmacology; D000199:Actins / Q000378:metabolism; D000273:Adipose Tissue / Q000166:cytology; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000201:enzymology; D009119:Muscle Contraction / Q000187:drug effects; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000187:drug effects* / Q000201:enzymology; D018994:Myosin Light Chains / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D017482:Receptors, Thromboxane / Q000378:metabolism; D026362:Serum Response Factor / Q000378:metabolism; D022502:Stress Fibers / Q000187:drug effects / Q000378:metabolism; D013928:Thromboxane A2 / Q000494:pharmacology*; D015534:Trans-Activators / Q000378:metabolism; D054460:rho-Associated Kinases / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism; D000099251:Myocardin
D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; D000199:Actins; D002118:Calcium; D018994:Myosin Light Chains; D009687:Nuclear Proteins; D017482:Receptors, Thromboxane; D026362:Serum Response Factor; D013928:Thromboxane A2; D015534:Trans-Activators; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein; D000099251:Myocardin