Puerarin facilitates T-tubule development of murine embryonic stem cell-derived cardiomyocytes

Cell Physiol Biochem. 2014;34(2):383-92. doi: 10.1159/000363007. Epub 2014 Jul 11.

Abstract

Aims: The embryonic stem cell-derived cardiomyocytes (ES-CM) is one of the promising cell sources for repopulation of damaged myocardium. However, ES-CMs present immature structure, which impairs their integration with host tissue and functional regeneration. This study used murine ES-CMs as an in vitro model of cardiomyogenesis to elucidate the effect of puerarin, the main compound found in the traditional Chinese medicine the herb Radix puerariae, on t-tubule development of murine ES-CMs.

Methods: Electron microscope was employed to examine the ultrastructure. The investigation of transverse-tubules (t-tubules) was performed by Di-8-ANEPPS staining. Quantitative real-time PCR was utilized to study the transcript level of genes related to t-tubule development.

Results: We found that long-term application of puerarin throughout cardiac differentiation improved myofibril array and sarcomeres formation, and significantly facilitated t-tubules development of ES-CMs. The transcript levels of caveolin-3, amphiphysin-2 and junctophinlin-2, which are crucial for the formation and development of t-tubules, were significantly upregulated by puerarin treatment. Furthermore, puerarin repressed the expression of miR-22, which targets to caveolin-3.

Conclusion: Our data showed that puerarin facilitates t-tubule development of murine ES-CMs. This might be related to the repression of miR-22 by puerarin and upregulation of Cav3, Bin1 and JP2 transcripts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / genetics
  • Animals
  • Base Sequence
  • Caveolin 3 / genetics
  • Cell Differentiation
  • DNA Primers
  • Embryonic Stem Cells / cytology*
  • Isoflavones / pharmacology*
  • Membrane Proteins / genetics
  • Mice
  • MicroRNAs / genetics
  • Microscopy, Electron, Transmission
  • Muscle Proteins / genetics
  • Myocytes, Cardiac / cytology
  • Myocytes, Cardiac / drug effects*
  • Myocytes, Cardiac / ultrastructure
  • Nerve Tissue Proteins / genetics
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Tumor Suppressor Proteins / genetics
  • Up-Regulation / drug effects

Substances

  • Adaptor Proteins, Signal Transducing
  • Bin1 protein, mouse
  • Caveolin 3
  • DNA Primers
  • Isoflavones
  • Membrane Proteins
  • MicroRNAs
  • Mirn22 microRNA, mouse
  • Muscle Proteins
  • Nerve Tissue Proteins
  • RNA, Messenger
  • Tumor Suppressor Proteins
  • junctophilin-2 protein, mouse
  • puerarin