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Thymosin β4 Impeded Murine Stem Cell Proliferation with an Intact Cardiovascular Differentiation

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【作者】 聂丽高仕君赵亚楠Jacob Masika骆红艳胡新武张亮品曾莹Jürgen Hescheler梁华敏

【Author】 Li NIE;Shi-jun GAO;Ya-nan ZHAO;Jacob Masika;Hong-yan LUO;Xin-wu HU;Liang-pin ZHANG;Ying ZENG;Jürgen Hescheler;Hua-min LIANG;Department of Physiology,Chinese-German Stem Cell Center,the Key Laboratory of Drug Target Research and Pharmacodynamic Evaluation,Hubei Province; School of Basic Medicine,Huazhong University of Science and Technology;Institute of Brain Research,Huazhong University of Science and Technology;College of Pharmacy,Wuhan Institute of Bioengineering;Department of Medical Physiology,Faculty of Health Sciences,Egerton University;Union Hospital,Huazhong University of Science and Technology;Institute of Neurophysiology,University of Cologne;

【机构】 Department of Physiology,Chinese-German Stem Cell Center,the Key Laboratory of Drug Target Research and Pharmacodynamic Evaluation,Hubei ProvinceSchool of Basic Medicine,Huazhong University of Science and TechnologyInstitute of Brain Research,Huazhong University of Science and TechnologyCollege of Pharmacy,Wuhan Institute of BioengineeringDepartment of Medical Physiology,Faculty of Health Sciences,Egerton UniversityUnion Hospital,Huazhong University of Science and TechnologyInstitute of Neurophysiology,University of Cologne

【摘要】 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(m ESCs) proliferation and cardiovascular differentiation remains unclear. Thus we aimed to elucidate the influence of Tβ4 on m ESCs. Target genes during m ESCs proliferation and differentiation were detected by real-time PCR or Western blotting, and patch clamp was applied to characterize the m ESCs-derived cardiomyocytes. It was found that Tβ4 decreased m ESCs proliferation in a partial dose-dependent manner and the expression of cell cycle regulatory genes c-myc, c-fos and c-jun. However, m ESCs self-renewal markers Oct4 and Nanog were elevated, indicating the maintenance of self-renewal ability in these m ESCs. 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 ?-catenin. Under m ESCs differentiation, Tβ4 treatment did not change the expression of cardiovascular cell markers α-MHC, PECAM, and α-SMA. Neither the electrophysiological properties of m ESCs-derived cardiomyocytes nor the hormonal regulation by Iso/Cch was affected by Tβ4. In conclusion, Tβ4 suppressed m ESCs proliferation by affecting the activity of STAT3, Akt, ERK and Wnt pathways. However, Tβ4 did not influence the in vitro 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(m ESCs) proliferation and cardiovascular differentiation remains unclear. Thus we aimed to elucidate the influence of Tβ4 on m ESCs. Target genes during m ESCs proliferation and differentiation were detected by real-time PCR or Western blotting, and patch clamp was applied to characterize the m ESCs-derived cardiomyocytes. It was found that Tβ4 decreased m ESCs proliferation in a partial dose-dependent manner and the expression of cell cycle regulatory genes c-myc, c-fos and c-jun. However, m ESCs self-renewal markers Oct4 and Nanog were elevated, indicating the maintenance of self-renewal ability in these m ESCs. 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 ?-catenin. Under m ESCs differentiation, Tβ4 treatment did not change the expression of cardiovascular cell markers α-MHC, PECAM, and α-SMA. Neither the electrophysiological properties of m ESCs-derived cardiomyocytes nor the hormonal regulation by Iso/Cch was affected by Tβ4. In conclusion, Tβ4 suppressed m ESCs proliferation by affecting the activity of STAT3, Akt, ERK and Wnt pathways. However, Tβ4 did not influence the in vitro cardiovascular differentiation.

【基金】 supposed by grants from National Natural Science Foundation of China(No.81100818,No.31100828 and No.81070342);the Fundamental Research Funds for the Central Universities(HUST:No.2012TS036)
  • 【文献出处】 Journal of Huazhong University of Science and Technology(Medical Sciences) ,华中科技大学学报(医学英德文版) , 编辑部邮箱 ,2016年03期
  • 【分类号】R329.2
  • 【被引频次】1
  • 【下载频次】20
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