Cardiac Protective Effect of Kirenol against Doxorubicin-Induced Cardiac Hypertrophy in H9c2 Cells through Nrf2 Signaling via PI3K/AKT Pathways

Int J Mol Sci. 2021 Mar 23;22(6):3269. doi: 10.3390/ijms22063269.


Kirenol (KRL) is a biologically active substance extracted from Herba Siegesbeckiae. This natural type of diterpenoid has been widely adopted for its important anti-inflammatory and anti-rheumatic properties. Despite several studies claiming the benefits of KRL, its cardiac effects have not yet been clarified. Cardiotoxicity remains a key concern associated with the long-term administration of doxorubicin (DOX). The generation of reactive oxygen species (ROS) causes oxidative stress, significantly contributing to DOX-induced cardiac damage. The purpose of the current study is to investigate the cardio-protective effects of KRL against apoptosis in H9c2 cells induced by DOX. The analysis of cellular apoptosis was performed using the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining assay and measuring the modulation in the expression levels of proteins involved in apoptosis and Nrf2 signaling, the oxidative stress markers. Furthermore, Western blotting was used to determine cell survival. KRL treatment, with Nrf2 upregulation and activation, accompanied by activation of PI3K/AKT, could prevent the administration of DOX to induce cardiac oxidative stress, remodeling, and other effects. Additionally, the diterpenoid enhanced the activation of Bcl2 and Bcl-xL, while suppressing apoptosis marker proteins. As a result, KRL is considered a potential agent against hypertrophy resulting from cardiac deterioration. The study results show that KRL not only activates the IGF-IR-dependent p-PI3K/p-AKT and Nrf2 signaling pathway, but also suppresses caspase-dependent apoptosis.

Keywords: H9c2; Nrf2; cardiotoxicity; doxorubicin; hypertrophy; kirenol.

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Cardiotonic Agents / pharmacology*
  • Cell Death / drug effects
  • Cell Line
  • Cell Survival / drug effects
  • Cytoskeleton / metabolism
  • Diterpenes / chemistry
  • Diterpenes / pharmacology*
  • Doxorubicin / adverse effects
  • Humans
  • Matrix Metalloproteinase 2 / metabolism
  • Matrix Metalloproteinase 9 / metabolism
  • Myoblasts, Cardiac / drug effects
  • Myoblasts, Cardiac / metabolism
  • Myocytes, Cardiac / drug effects
  • Myocytes, Cardiac / metabolism
  • Myocytes, Cardiac / pathology
  • NF-E2-Related Factor 2 / metabolism*
  • Natriuretic Peptides / metabolism
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Phosphorylation
  • Protein Transport
  • Proto-Oncogene Proteins c-akt / metabolism*
  • Signal Transduction / drug effects*


  • Cardiotonic Agents
  • Diterpenes
  • NF-E2-Related Factor 2
  • Natriuretic Peptides
  • kirenol
  • Doxorubicin
  • Proto-Oncogene Proteins c-akt
  • Matrix Metalloproteinase 2
  • Matrix Metalloproteinase 9