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Icariin promotes expression of PGC-1α, PPARα, and NRF-1 during cardiomyocyte differentiation of murine embryonic stem cells in vitro

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Abstract

Aim: To investigate the effect of icariin on the expression of peroxisome proliferator-activated receptor  coactivator-1 alpha (PGC-1α), peroxisome proliferator-activated receptor alpha (PPARα), and nuclear respiratory factor 1 (NRF-1) on cardiomyocyte differentiation of murine embryonic stem (ES) cells in vitro.Methods: The cardiomyocytes derived from murine ES cells were verified by immunocytochemistry using confocal laser scanning microscopy. Cardiac-specific sarcomeric proteins (ie α-actinin, troponin T) were evaluated when embryoid bodies (EB) were treated with icariin or retinoid acid. The expression of PGC-1α, PPARα, and NRF-1 were analyzed using both semiquantitative RT-PCR and Western blotting in cardiomyocyte differentiation. The phosphorylation of the p38 mitogen-activated protein kinase (MAPK) was studied in the differentiation process, and its specific inhibitor SB203580 was employed to confirm the function of the p38 MAPK on icariin-induced cardiac differentiation. Results: The application of icariin significantly induced the cardiomyocyte differentiation of EB as indicated by the promoted expression of α-actinin and troponin T. The expression of PGC-1α, PPARα, and NRF-1 increased coincidently in early differentiation and the increase was dose-dependently upregulated by icariin treatment. The phosphorylation of the p38 MAPK peaked on d 6 and decreased after d 8, and the activation was further enhanced and prolonged when the EB were subjected to icariin, which was concurrent with the elevation of PGC-1α, PPARα, and NRF-1. Moreover, the inhibition of the p38 MAPK pathway by SB203580 efficiently abolished icariin-stimulated cardiomyocyte differentiation and resulted in the capture of the upregulation of PGC-1α, PPARα, and NRF-1. Conclusion: Taken together, icariin promoted the expression of PGC-1α, PPARα, and NRF-1 during cardiomyocyte differentiation of murine ES cells in vitro and the effect was partly responsible for the activation of the p38 MAPK.
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Keywords: NRF-1; PGC-1α; PPARα; cardiomyocytes; differentiation; embryonic stem cells; icariin; mitochondrial

Document Type: Research Article

Affiliations: Institute of Pharmacology and Toxicology and Biochemical Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China

Publication date: 2007-10-01

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