Involvement of PIKE in icariin induced cardiomyocyte differentiation from murine embryonic stem cells.
Icariin (ICA) has demonstrated to induce cardiomyocyte differentiation from murine embryonic stem (ES) cells in vitro, however, the mechanisms have not been fully elucidated. In the present study, we investigated whether phosphatidylinositol 3-kinase enhancer (PIKE) was involved in ICA induced cardiomyocyte differentiation of ES cells. Small interfering RNA (siRNA) of PIKE was applied to investigate the role of PIKE in ICA induced cardiomyocyte differentiation. The cardiomyocytes derived from ES cells were verified using immunofluorescence. The expressions of Troponin T, PIKE, phosphatidylinositol 3-kinase (PI3K), and nuclear factor-kappaB (NF-kappaB) were detected by western blot. The change of reactive oxygen species (ROS) generation was estimated using the fluorescent dye 2', 7' - dichlorodihydrofluorescein diacetate. The results showed that PIKE expression increased during cardiomyocyte differentiation. ICA markedly enhanced PIKE and PI3K expression in a time-dependent manner. Knockdown of PIKE by siRNAs blocked the differentiation of ES cells into cardiomyocytes expressing alpha-actinin for cardiac sarcomeric structures. Moreover, reduced ROS generation and NF-kappaB nuclear translocation were responsible for the inhibitory effect of si-PIKE. In conclusion, PIKE was involved in ICA induced cardiomyocyte differentiation, and ROS generation and NF-kappaB nuclear translocation were associated with PIKE activation.
Limin
L
Zhou
Bei
B
Zheng
Leilei
L
Tang
Yujie
Y
Huang
Danyan
D
Zhu
D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000187:drug effects*; D005419:Flavonoids / Q000494:pharmacology*; D005455:Fluorescent Antibody Technique; D020558:GTP Phosphohydrolases / Q000378:metabolism*; D051379:Mice; D032383:Myocytes, Cardiac / Q000187:drug effects*; D016328:NF-kappa B / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D014178:Translocation, Genetic / Q000187:drug effects
D005419:Flavonoids; D016328:NF-kappa B; D009419:Nerve Tissue Proteins; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D020558:GTP Phosphohydrolases; C529781:PIKE protein, mouse; C056599:icariin