TY - JOUR
T1 - Regulation of multiple transcription factors by reactive oxygen species and effects of pro-inflammatory cytokines released during myocardial infarction on cardiac differentiation of embryonic stem cells
AU - Law, Sau Kwan
AU - Leung, Cecilia Sze Lee
AU - Yau, Ka Long
AU - Tse, Chi Lok
AU - Wong, Chun Kit
AU - LEUNG, Fung Ping
AU - Mascheck, Lena
AU - Huang, Yu
AU - Sauer, Heinrich
AU - Tsang, Suk Ying
N1 - Funding Information:
This study was supported by the Competitive Earmarked Research Grant ( 474907 ) from the University Grants Committee (UGC) of the Hong Kong SAR , the Germany/Hong Kong Joint Research Scheme ( G_HK029/11 ) from the Deutsche Akademische Austauschdienst and the UGC, and Innovative Technology Fund of Innovation Technology Commission: Funding Support to Partner State Key Laboratories in Hong Kong. S.K.L., C.S.L., K.L.Y. and C.K.W. were supported by postgraduate studentships from the CUHK . We would like to thank Dr. Winnie Poon for her excellent technical support.
PY - 2013/10/9
Y1 - 2013/10/9
N2 - Background The mechanism of how reactive oxygen species (ROS) regulate cardiac differentiation in the long-run is unclear and the effect of pro-inflammatory cytokines secreted during myocardial infarction on the cardiac differentiation of embryonic stem cells (ESCs) is unknown. The aims of this study were 1) to investigate the effect of ROS on cardiac differentiation and the regulations of transcription factors in ESC differentiation cultures and 2) to investigate the effect of pro-inflammatory cytokines on the expression of cardiac structural genes and whether this effect is mediated through ROS signaling. Methods ESCs were differentiated using hanging drop method. Degree of cardiac differentiation was determined by the appearance of beating embryoid bodies (EBs) and by the expression of cardiac genes using real-time PCR and Western blot. Intracellular ROS level was examined by confocal imaging. Results H2O2-treated EBs were found to have enhanced cardiac differentiation in the long run as reflected by, firstly, an earlier appearance of beating EBs, and secondly, an upregulation in cardiac structural protein expression at both mRNA and protein levels. Also, ROS upregulated the expression of several cardiac-related transcription factors, and increased the post-translationally-activated transcription factors SRF and AP-1. IL-1β, IL-10, IL-18 and TNF-α upregulated the expression of cardiac structural proteins and increased the ROS level in differentiating EBs. In addition, ROS scavenger reversed the cardiogenic effect of IL-10 and IL-18. Conclusions These results demonstrated that ROS enhance cardiac differentiation of ESCs through upregulating the expression and activity of multiple cardiac-related transcription factors. IL-1β, IL-10, IL-18 and TNF-α enhance cardiac differentiation and ROS may serve as the messenger in cardiogenic signaling from these cytokines.
AB - Background The mechanism of how reactive oxygen species (ROS) regulate cardiac differentiation in the long-run is unclear and the effect of pro-inflammatory cytokines secreted during myocardial infarction on the cardiac differentiation of embryonic stem cells (ESCs) is unknown. The aims of this study were 1) to investigate the effect of ROS on cardiac differentiation and the regulations of transcription factors in ESC differentiation cultures and 2) to investigate the effect of pro-inflammatory cytokines on the expression of cardiac structural genes and whether this effect is mediated through ROS signaling. Methods ESCs were differentiated using hanging drop method. Degree of cardiac differentiation was determined by the appearance of beating embryoid bodies (EBs) and by the expression of cardiac genes using real-time PCR and Western blot. Intracellular ROS level was examined by confocal imaging. Results H2O2-treated EBs were found to have enhanced cardiac differentiation in the long run as reflected by, firstly, an earlier appearance of beating EBs, and secondly, an upregulation in cardiac structural protein expression at both mRNA and protein levels. Also, ROS upregulated the expression of several cardiac-related transcription factors, and increased the post-translationally-activated transcription factors SRF and AP-1. IL-1β, IL-10, IL-18 and TNF-α upregulated the expression of cardiac structural proteins and increased the ROS level in differentiating EBs. In addition, ROS scavenger reversed the cardiogenic effect of IL-10 and IL-18. Conclusions These results demonstrated that ROS enhance cardiac differentiation of ESCs through upregulating the expression and activity of multiple cardiac-related transcription factors. IL-1β, IL-10, IL-18 and TNF-α enhance cardiac differentiation and ROS may serve as the messenger in cardiogenic signaling from these cytokines.
KW - Cardiac differentiation
KW - Cytokines
KW - Embryonic stem cells
KW - Reactive oxygen species
KW - Transcriptional factors
UR - http://www.scopus.com/inward/record.url?scp=84886256432&partnerID=8YFLogxK
U2 - 10.1016/j.ijcard.2013.04.178
DO - 10.1016/j.ijcard.2013.04.178
M3 - Journal article
C2 - 23706318
AN - SCOPUS:84886256432
SN - 0167-5273
VL - 168
SP - 3458
EP - 3472
JO - International Journal of Cardiology
JF - International Journal of Cardiology
IS - 4
ER -