Objective:

2

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:

2

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 cleavedcaspase 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:

2

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.

Conclusion:

2

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.