title: Thymosin β4 impeded murine stem cell proliferation with an intact cardiovascular differentiation.
abstract: Thymosin β4 (Tβ4) is a key factor in cardiac development, growth, disease, epicardial integrity, blood vessel formation and has cardio-protective properties. However, its role in murine embryonic stem cells (mESCs) proliferation and cardiovascular differentiation remains unclear. Thus we aimed to elucidate the influence of Tβ4 on mESCs. Target genes during mESCs proliferation and differentiation were detected by real-time PCR or Western blotting, and patch clamp was applied to characterize the mESCs-derived cardiomyocytes. It was found that Tβ4 decreased mESCs proliferation in a partial dose-dependent manner and the expression of cell cycle regulatory genes c-myc, c-fos and c-jun. However, mESCs self-renewal markers Oct4 and Nanog were elevated, indicating the maintenance of self-renewal ability in these mESCs. Phosphorylation of STAT3 and Akt was inhibited by Tβ4 while the expression of RAS and phosphorylation of ERK were enhanced. No significant difference was found in BMP2/BMP4 or their downstream protein smad. Wnt3 and Wnt11 were remarkably decreased by Tβ4 with upregulation of Tcf3 and constant β-catenin. Under mESCs differentiation, Tβ4 treatment did not change the expression of cardiovascular cell markers α-MHC, PECAM, and α-SMA. Neither the electrophysiological properties of mESCs-derived cardiomyocytes nor the hormonal regulation by Iso/Cch was affected by Tβ4. In conclusion, Tβ4 suppressed mESCs proliferation by affecting the activity of STAT3, Akt, ERK and Wnt pathways. However, Tβ4 did not influence the in vitro cardiovascular differentiation.
authors.affiliation: Department of Physiology, Chinese-German Stem Cell Center, the Key Laboratory of Drug Target Research and Pharmacodynamic Evaluation, Hubei Province, Huazhong University of Science and Technology, Wuhan, 430030, China.|Institute of Brain Research, Huazhong University of Science and Technology, Wuhan, 430030, China.|College of Pharmacy, Wuhan Institute of Bioengineering, Wuhan, 430415, China.|School of Basic Medicine, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Li
authors.initials: L
authors.lastname: Nie
authors.affiliation: Department of Physiology, Chinese-German Stem Cell Center, the Key Laboratory of Drug Target Research and Pharmacodynamic Evaluation, Hubei Province, Huazhong University of Science and Technology, Wuhan, 430030, China.|Institute of Brain Research, Huazhong University of Science and Technology, Wuhan, 430030, China.|School of Basic Medicine, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Shi-Jun
authors.initials: SJ
authors.lastname: Gao
authors.affiliation: Department of Physiology, Chinese-German Stem Cell Center, the Key Laboratory of Drug Target Research and Pharmacodynamic Evaluation, Hubei Province, Huazhong University of Science and Technology, Wuhan, 430030, China.|Institute of Brain Research, Huazhong University of Science and Technology, Wuhan, 430030, China.|School of Basic Medicine, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Ya-Nan
authors.initials: YN
authors.lastname: Zhao
authors.affiliation: Department of Physiology, Chinese-German Stem Cell Center, the Key Laboratory of Drug Target Research and Pharmacodynamic Evaluation, Hubei Province, Huazhong University of Science and Technology, Wuhan, 430030, China.|Institute of Brain Research, Huazhong University of Science and Technology, Wuhan, 430030, China.|Department of Medical Physiology, Faculty of Health Sciences, Egerton University, Nakura, 53620115, Kenya.|School of Basic Medicine, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Jacob
authors.initials: J
authors.lastname: Masika
authors.affiliation: Department of Physiology, Chinese-German Stem Cell Center, the Key Laboratory of Drug Target Research and Pharmacodynamic Evaluation, Hubei Province, Huazhong University of Science and Technology, Wuhan, 430030, China.|Institute of Brain Research, Huazhong University of Science and Technology, Wuhan, 430030, China.|School of Basic Medicine, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Hong-Yan
authors.initials: HY
authors.lastname: Luo
authors.affiliation: Department of Physiology, Chinese-German Stem Cell Center, the Key Laboratory of Drug Target Research and Pharmacodynamic Evaluation, Hubei Province, Huazhong University of Science and Technology, Wuhan, 430030, China.|Institute of Brain Research, Huazhong University of Science and Technology, Wuhan, 430030, China.|School of Basic Medicine, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Xin-Wu
authors.initials: XW
authors.lastname: Hu
authors.affiliation: Department of Physiology, Chinese-German Stem Cell Center, the Key Laboratory of Drug Target Research and Pharmacodynamic Evaluation, Hubei Province, Huazhong University of Science and Technology, Wuhan, 430030, China.|Institute of Brain Research, Huazhong University of Science and Technology, Wuhan, 430030, China.|School of Basic Medicine, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Liang-Pin
authors.initials: LP
authors.lastname: Zhang
authors.affiliation: Union Hospital, Huazhong University of Science and Technology, Wuhan, 430022, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zeng
authors.affiliation: Institute of Neurophysiology, University of Cologne, Robert Koch str. 39, Cologne, 50931, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Hescheler
authors.affiliation: Department of Physiology, Chinese-German Stem Cell Center, the Key Laboratory of Drug Target Research and Pharmacodynamic Evaluation, Hubei Province, Huazhong University of Science and Technology, Wuhan, 430030, China. lianghuamin76@163.com.|Institute of Brain Research, Huazhong University of Science and Technology, Wuhan, 430030, China. lianghuamin76@163.com.|School of Basic Medicine, Huazhong University of Science and Technology, Wuhan, 430030, China. lianghuamin76@163.com.
authors.forename: Hua-Min
authors.initials: HM
authors.lastname: Liang
mesh_terms: D000818:Animals; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D048031:JNK Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D000071317:Nanog Homeobox Protein / Q000235:genetics / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D018408:Patch-Clamp Techniques; D061251:Primary Cell Culture; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D016760:Proto-Oncogene Proteins c-fos / Q000235:genetics / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D013947:Thymosin / Q000494:pharmacology*; D000099201:Thymosin beta(4)
chemical_list: D048049:Extracellular Signal-Regulated MAP Kinases; D048031:JNK Mitogen-Activated Protein Kinases; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; D051057:Proto-Oncogene Proteins c-akt; D016760:Proto-Oncogene Proteins c-fos; D016271:Proto-Oncogene Proteins c-myc; D050796:STAT3 Transcription Factor; D013947:Thymosin; D000099201:Thymosin beta(4); C489428:Myc protein, mouse; C475540:Nanog protein, mouse; C494174:Pou5f1 protein, mouse; C494087:Stat3 protein, mouse
keywords: cardiogenesis; murine embryonic stem cells; patch clamp; proliferation; thymosin β4