摘要
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/m L 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(m PTP), 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 cel s 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 m PTP. 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: 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.
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/m L 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(m PTP), 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 cel s 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 m PTP. 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: 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.
基金
Supported by the National Basic Research Program of China(973 Program,No.2015CB554400)
