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Apoptosis

, Volume 21, Issue 8, pp 887–895 | Cite as

Canstatin inhibits isoproterenol-induced apoptosis through preserving mitochondrial morphology in differentiated H9c2 cardiomyoblasts

  • Muneyoshi Okada
  • Suiri Morioka
  • Hiroki Kanazawa
  • Hideyuki Yamawaki
Article

Abstract

Canstatin, a non-collagenous fragment, is cleaved from type IV collagen α2 chain, an essential component of basement membrane surrounding cardiomyocytes. Although canstatin is known as an endogenous anti-angiogenic factor, its effects on cardiomyocytes have not been clarified. This study examined the effects of canstatin on isoproterenol-induced apoptosis in differentiated H9c2 cardiomyoblasts. Retinoic acid was used to differentiate H9c2 myoblast to cardiomyocyte-like phenotype. Cell viability was determined by a cell counting assay. Western blotting was performed to detect expression of cleaved casepase-3 and phosphorylation of dynamin related protein (Drp)1 at Ser637 which regulates mitochondrial fission. Mito Sox Red staining was performed to examine a mitochondria-dependent production of reactive oxygen species (ROS). Mitochondrial morphology was detected by Mito Tracker Red staining. Isoproterenol (100 μM, 48 h) significantly decreased cell viability and increased cleaved caspase-3 expression, which were inhibited by canstatin (10–250 ng/ml) in a concentration-dependent manner. Canstatin suppressed the isoproterenol-induced mitochondrial fission but not ROS. Canstatin also inhibited the isoproterenol-induced dephosphorylation of Drp1 at Ser637. In conclusion, canstatin inhibits isoproterenol-induced apoptosis through the inhibition of mitochondrial fission via the suppression of dephosphorylation of Drp1 at Ser637 in differentiated H9c2 cardiomyoblasts.

Keywords

Apoptosis Canstatin Dynamin related protein 1 Isoproterenol Mitochondrial fission 

Abbreviations

DMEM

Dulbecco’s modified Eagle’s Medium

Drp1

Dynamin related protein 1

NFAT

Nuclear factor of activated T-cells

PI3K

Phosphatidylinositol 3-kinase

ROS

Reactive oxygen species

S.E.M.

Standard error of the mean

Notes

Acknowledgments

This research was supported by Kitasato University Research Grant for Young Researchers and JSPS KAKENHI Grant Number 24780289 (Grant-in-Aid for Young Scientists B).

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Muneyoshi Okada
    • 1
  • Suiri Morioka
    • 1
  • Hiroki Kanazawa
    • 1
  • Hideyuki Yamawaki
    • 1
  1. 1.Laboratory of Veterinary Pharmacology, School of Veterinary MedicineKitasato UniversityTowada, AomoriJapan

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