Protective effects of higenamine combined with [6]-gingerol against doxorubicin-induced mitochondrial dysfunction and toxicity in H9c2 cells and potential mechanisms

Biomed Pharmacother. 2019 Jul;115:108881. doi: 10.1016/j.biopha.2019.108881. Epub 2019 Apr 24.


Higenamine (HG) is a well-known selective activator of beta2-adrenergic receptor (β2-AR) with a positive inotropic effect. The present study showed that HG combined with [6]-gingerol (HG/[6]-GR) protects H9c2 cells from doxorubicin (DOX)-induced mitochondrial energy metabolism disorder and respiratory dysfunction. H9c2 cells were pretreated with HG/[6]-GR for 2 h before DOX treatment in all procedures. Cell viability was quantified by a cell counting kit‑8 assay. Cardiomyocyte morphology, proliferation, and mitochondrial function were detected by a high content screening (HCS) assay. Cell mitochondrial stress was measured by a Seahorse XFp analyzer. To further investigate the protective mechanism of HG/[6]-GR, mRNA and protein expression levels of PPARα/PGC-1α/Sirt3 pathway-related molecules were detected. The present data demonstrated that protective effects of HG/[6]-GR combination were presented in mitochondria, which increased cell viability, ameliorated DOX-induced mitochondrial dysfunction, increased mitochondrial oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). Most importantly, the protective effects were abrogated by GW6471 (a PPARα inhibitor) and ameliorated by Wy14643 (a PPARα agonist). Moreover, the combined use of HG and [6]-GR exerted more profound protective effects than either drug as a single agent. In conclusion, the results suggested that HG/[6]-GR ameliorates DOX-induced mitochondrial energy metabolism disorder and respiratory function impairment in H9c2 cells, and it indicated that the protective mechanism may be related to upregulation of the PPARα/PGC-1α/Sirt3 pathway, which promotes mitochondrial energy metabolism and protects against heart failure.

Keywords: Doxorubicin; Energy metabolism; H9c2 cells; Heart failure; Higenamine; [6]-Gingerol.

MeSH terms

  • Adenine Nucleotide Translocator 1 / genetics
  • Adenine Nucleotide Translocator 1 / metabolism
  • Adrenergic beta-Agonists / administration & dosage
  • Adrenergic beta-Agonists / pharmacology
  • Alkaloids / administration & dosage
  • Alkaloids / pharmacology*
  • Animals
  • Antibiotics, Antineoplastic / toxicity
  • Catechols / administration & dosage
  • Catechols / pharmacology*
  • Cell Line
  • Cell Survival
  • Doxorubicin / toxicity*
  • Energy Metabolism / drug effects
  • Fatty Alcohols / administration & dosage
  • Fatty Alcohols / pharmacology*
  • Gene Expression Regulation / drug effects
  • Mitochondria / drug effects*
  • Myocytes, Cardiac / drug effects*
  • PPAR alpha / genetics
  • PPAR alpha / metabolism
  • Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha / genetics
  • Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats
  • Sirtuins / genetics
  • Sirtuins / metabolism
  • Tetrahydroisoquinolines / administration & dosage
  • Tetrahydroisoquinolines / pharmacology*
  • p300-CBP Transcription Factors / genetics
  • p300-CBP Transcription Factors / metabolism


  • Adenine Nucleotide Translocator 1
  • Adrenergic beta-Agonists
  • Alkaloids
  • Antibiotics, Antineoplastic
  • Catechols
  • Fatty Alcohols
  • PPAR alpha
  • Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha
  • Ppargc1a protein, rat
  • RNA, Messenger
  • SIRT3 protein, rat
  • Tetrahydroisoquinolines
  • Doxorubicin
  • gingerol
  • p300-CBP Transcription Factors
  • Sirtuins
  • higenamine