title: Puerarin Exerts a Delayed Inhibitory Effect on the Proliferation of Cardiomyocytes Derived from Murine ES Cells via Slowing Progression through G2/M Phase.
abstract: OBJECTIVE
  Puerarin, which shows beneficial and protective effects on cardiovascular diseases, is the main isoflavone extracted from Pueraria lobata (kudzu) root. The aim of this study was to investigate the effects of puerarin on in vitro myocardial proliferation and its underlying mechanism.
  
  
  METHODS
  Myocardial differentiation of transgenic embryonic stem (ES) cells was performed by embryoid body-based differentiation method. The proliferation assay of cardiomyocytes (CMs) derived from ES cells (ES-CMs) was performed by EdU (5-Ethynyl-2'-deoxyuridine) staining. Flow cytometry was employed to determine the cell cycle distribution and apoptosis of purified ES-CMs. Quantitative real-time PCR was utilized to study the transcription of genes related to cell cycle progression. Signaling pathways relating to proliferation were studied by western blot analysis and application of specific inhibitors.
  
  
  RESULTS
  Puerarin exerted a delayed inhibitory effect on the proliferation of ES-CMs at the early-stage differentiation. Meanwhile, puerarin slowed progression through G2/M phase without inducing apoptosis of ES-CMs. Further assays showed that puerarin up-regulated the transcription of Cyclin A2, Cyclin B1 and Cdk1 in ES-CMs. The ERK1/2 specific inhibitor PD0325901 and the PI3K specific inhibitor Wortmannin successfully reversed puerarin-induced up-regulation of Cdk1 but not Cyclin A2 and B1.
  
  
  CONCLUSIONS
  These findings suggest that puerarin inhibits CM proliferation via slowing progression through G2/M phase during early-stage differentiation.
authors.forename: Xueying
authors.initials: X
authors.lastname: Luo
authors.forename: Binlong
authors.initials: B
authors.lastname: Zhong
authors.forename: Xian
authors.initials: X
authors.lastname: Hong
authors.forename: Yurong
authors.initials: Y
authors.lastname: Cui
authors.forename: Ying
authors.initials: Y
authors.lastname: Gao
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Yin
authors.forename: Ming
authors.initials: M
authors.lastname: Tang
authors.forename: Jürgen
authors.initials: J
authors.lastname: Hescheler
authors.forename: Jiaoya
authors.initials: J
authors.lastname: Xi
mesh_terms: D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D016203:CDC2 Protein Kinase / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D056751:Cyclin A2 / Q000235:genetics / Q000378:metabolism; D056744:Cyclin B1 / Q000235:genetics / Q000378:metabolism; D004159:Diphenylamine / Q000031:analogs & derivatives / Q000494:pharmacology; D059565:G2 Phase Cell Cycle Checkpoints / Q000187:drug effects; D007529:Isoflavones / Q000494:pharmacology*; D059566:M Phase Cell Cycle Checkpoints / Q000187:drug effects; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism; D018517:Plant Roots / Q000737:chemistry / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011494:Protein Kinases / Q000378:metabolism; D029908:Pueraria / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D015854:Up-Regulation / Q000187:drug effects; D014665:Vasodilator Agents / Q000494:pharmacology*; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D001549:Benzamides; D056751:Cyclin A2; D056744:Cyclin B1; D007529:Isoflavones; D047428:Protein Kinase Inhibitors; D014665:Vasodilator Agents; C506614:mirdametinib; D004159:Diphenylamine; D011494:Protein Kinases; D016203:CDC2 Protein Kinase; D000077191:Wortmannin; C033607:puerarin