Astragaloside IV inhibits doxorubicin-induced cardiomyocyte apoptosis mediated by mitochondrial apoptotic pathway via activating the PI3K/Akt pathway

Chem Pharm Bull (Tokyo). 2014;62(1):45-53. doi: 10.1248/cpb.c13-00556.


Doxorubicin (DOX) is a widely used antitumor drug whose application is seriously limited by its cardiotoxicity. Mitochondria-mediated cardiomyocyte apoptosis plays a critical role in DOX-induced cardiotoxicity (DIC). The aim of the present study was to investigate the protective effect of astragaloside IV (3-O-beta-D-xylopyranosyl-6-O-beta-D-glucopyranosyl-cycloastragenol, AS-IV), a pure saponin isolated from Astragalus membranaceus, against DOX-induced cardiomyocyte apoptosis in primary cultured neonatal rat cardiomyocytes. Immunocytochemistry and Microculture Tetrazolium (MTT) assays showed that AS-IV significantly reduced DOX-induced cardiomyocyte loss. Additionally, AS-IV markedly ameliorated DOX-caused cardiomyocyte dysfunction via restoring the beating cell ratio and beating rate in cardiomyocytes. Furthermore, AS-IV substantially reduced the mitochondrial reactive oxygen species (ROS) production and lactate dehydrogenase (LDH), creatine kinase-MB isoenzyme (CK-MB) and cytochrome c (CytC) release, and restored the reduced ATP level, succinate dehydrogenase (SDH) and ATP synthase activities induced by DOX, suggesting that AS-IV significantly attenuated DOX-induced mitochondrial damage and dysfunction. It was further observed that DOX-induced cardiomyocyte apoptosis, as qualitatively evaluated by Hoechst 33258 staining and accurately quantified by flow cytometry, was markedly inhibited by AS-IV. Western blot analysis manifested that AS-IV significantly inhibited the activation of mitochondrial apoptotic pathway (MAP) via inducing the phosphorylation of Akt and Bad. Furthermore, phosphatidylinositol 3-kinase (PI3K) inhibitor 2-(4-morpholinyl)-8-phenyl-1(4H)-benzopyran-4-one hydrochloride (LY294002) remarkably inhibited the anti-apoptotic effect of AS-IV. Moreover, AS-IV didn't compromise the antitumor activity of DOX. Taken together, our findings indicate that AS-IV ameliorates DIC, and this beneficial effect appears to be dependent on the activation of the PI3K/Akt pathway. Thus, AS-IV may hold promise as an efficient cardioprotective agent against DIC.

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Animals
  • Apoptosis / drug effects*
  • Cells, Cultured
  • Creatine Kinase, Mitochondrial Form / metabolism
  • Cytochromes c / metabolism
  • Doxorubicin / adverse effects*
  • L-Lactate Dehydrogenase / metabolism
  • Mitochondria / drug effects*
  • Mitochondria / metabolism
  • Mitochondrial Proton-Translocating ATPases / metabolism
  • Myocytes, Cardiac / drug effects*
  • Myocytes, Cardiac / metabolism
  • Phosphatidylinositol 3-Kinase / metabolism*
  • Phosphorylation / drug effects
  • Proto-Oncogene Proteins c-akt / metabolism*
  • Rats
  • Reactive Oxygen Species / metabolism
  • Saponins / pharmacology*
  • Succinate Dehydrogenase / metabolism
  • Triterpenes / pharmacology*
  • bcl-Associated Death Protein / metabolism


  • Bad protein, rat
  • Reactive Oxygen Species
  • Saponins
  • Triterpenes
  • bcl-Associated Death Protein
  • astragaloside A
  • Doxorubicin
  • Adenosine Triphosphate
  • Cytochromes c
  • L-Lactate Dehydrogenase
  • Succinate Dehydrogenase
  • Phosphatidylinositol 3-Kinase
  • Proto-Oncogene Proteins c-akt
  • Creatine Kinase, Mitochondrial Form
  • Mitochondrial Proton-Translocating ATPases