title: Salvianolic Acid A Protects Neonatal Cardiomyocytes Against Hypoxia/Reoxygenation-Induced Injury by Preserving Mitochondrial Function and Activating Akt/GSK-3β Signals.
abstract: OBJECTIVE
  To investigate the effects of salvianolic acid A (SAA) on cardiomyocyte apoptosis and mitochondrial dysfunction in response to hypoxia/reoxygenation (H/R) injury and to determine whether the Akt signaling pathway might play a role.
  
  
  METHODS
  An in vitro model of H/R injury was used to study outcomes on primary cultured neonatal rat cardiomyocytes. The cardiomyocytes were treated with 12.5, 25, 50 μg/mL SAA at the beginning of hypoxia and reoxygenation, respectively. Adenosine triphospate (ATP) and reactive oxygen species (ROS) levels were assayed. Cell apoptosis was evaluated by flow cytometry and the expression of cleaved-caspase 3, Bax and Bcl-2 were detected by Western blotting. The effects of SAA on mitochondrial dysfunction were examined by determining the mitochondrial membrane potential (△Ψm) and mitochondrial permeability transition pore (mPTP), followed by the phosphorylation of Akt (p-Akt) and GSK-3β (p-GSK-3β), which were measured by Western blotting.
  
  
  RESULTS
  SAA significantly preserved ATP levels and reduced ROS production. Importantly, SAA markedly reduced the number of apoptotic cells and decreased cleaved-caspase 3 expression levels, while also reducing the ratio of Bax/Bcl-2. Furthermore, SAA prevented the loss of △Ψm and inhibited the activation of mPTP. Western blotting experiments further revealed that SAA significantly increased the expression of p-Akt and p-GSK-3β, and the increase in p-GSK-3β expression was attenuated after inhibition of the Akt signaling pathway with LY294002.
  
  
  CONCLUSIONS
  SAA has a protective effect on cardiomyocyte H/R injury; the underlying mechanism may be related to the preservation of mitochondrial function and the activation of the Akt/GSK-3β signaling pathway.
authors.affiliation: Institute of Basic Medical Sciences, Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, 100091, China.|Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing, 100700, China.|Post-doctoral Research Station, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
authors.forename: Xue-Li
authors.initials: XL
authors.lastname: Li
authors.affiliation: China Academy of Chinese Medical Sciences, Beijing, 100700, China.|Eye Function Laboratory, Yanke Hospital, China Academy of Chinese Medical Sciences, Beijing, 100040, China.
authors.forename: Ji-Ping
authors.initials: JP
authors.lastname: Fan
authors.affiliation: Institute of Basic Medical Sciences, Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, 100091, China. liujx0324@sina.com.
authors.forename: Jian-Xun
authors.initials: JX
authors.lastname: Liu
authors.affiliation: Eye Function Laboratory, Yanke Hospital, China Academy of Chinese Medical Sciences, Beijing, 100040, China.
authors.forename: Li-Na
authors.initials: LN
authors.lastname: Liang
mesh_terms: D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D000831:Animals, Newborn; D002109:Caffeic Acids / Q000494:pharmacology*; D015687:Cell Hypoxia; D002478:Cells, Cultured; D000071679:Glycogen Synthase Kinase 3 beta / Q000502:physiology*; D007773:Lactates / Q000494:pharmacology*; D008929:Mitochondria, Heart / Q000187:drug effects* / Q000502:physiology; D033681:Mitochondrial Membrane Transport Proteins / Q000187:drug effects; D000083162:Mitochondrial Permeability Transition Pore; D032383:Myocytes, Cardiac / Q000187:drug effects; D051057:Proto-Oncogene Proteins c-akt / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D002109:Caffeic Acids; D007773:Lactates; D033681:Mitochondrial Membrane Transport Proteins; D000083162:Mitochondrial Permeability Transition Pore; D017382:Reactive Oxygen Species; C066201:salvianolic acid A; D000255:Adenosine Triphosphate; D000071679:Glycogen Synthase Kinase 3 beta; D051057:Proto-Oncogene Proteins c-akt
keywords: Akt/GSK-3β; cardiomyocyte; hypoxia/reoxygenation injury; mitochondria; salvianolic acid A