Kaempferol protects cardiomyocytes against anoxia/reoxygenation injury via mitochondrial pathway mediated by SIRT1.

Mitochondria-mediated apoptosis is a critical mechanism of anoxia/ reoxygenation (A/R)-induced injury in cardiomyocytes. Kaempferol (Kae) is a natural polyphenol and a type of flavonoid, which has been demonstrated to protect myocardium against ischemia/reperfusion (I/R) injury. However, the mechanism is still not fully elucidated. We hypothesize that Kae may improve the mitochondrial function during I/R injury via a potential signal pathway. In this study, an in vitro I/R model was replicated on neonatal rat primary cardiomyocytes by A/R treatment. Cell viability was monitored by the 3-(4,5-dimethylthiazol- 2-yl)-5-(3- carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 H-tetrazolium (MTS) assay. The levels of intracellular reactive oxygen species, mitochondrial membrane potential (Δψm) and apoptosis were determined by flow cytometry. Protein expression was detected by Western Blotting. mPTP opening and the activity of caspase-3 were measured by colorimetric method. The results showed that Kae effectively enhanced the cell viability and decreased the LDH release in cardiomyocytes subjected to A/R injury. Kae reduced the A/R-induced reactive oxygen species generation, the loss of Δψm, and the release of cytochrome c from mitochondria into cytosol. Kae inhibited the A/R-stimulated mPTP opening and activation of caspase-3, and ultimate decrease in cardiomyocytes apoptosis. Furthermore, we found Kae up-regulated Human Silent Information Regulator Type 1 (SIRT1) expression, indicating SIRT1 signal pathway likely involved the cardioprotection of Kae. Sirtinol, a SIRT1 inhibitor, abolished the protective effect of Kae in cardiomyocytes subjected to A/R. Additionally, Kae significantly increased the expression of Bcl-2. Thus, we firstly demonstrate that Kae protects cardiomyocytes against A/R injury through mitochondrial pathway mediated by SIRT1.

Guo Z ，Liao Z ，Huang L ，Liu D ，Yin D ，He M ... -  《-》

Protective Effects of Isorhamnetin on Cardiomyocytes Against Anoxia/Reoxygenation-induced Injury Is Mediated by SIRT1.

Huang L ，He H ，Liu Z ，Liu D ，Yin D ，He M ... -  《-》

Ganoderma atrum polysaccharide protects cardiomyocytes against anoxia/reoxygenation-induced oxidative stress by mitochondrial pathway.

It is now well established that oxidative stress plays a causative role in the pathogenesis of anoxia/reoxygenation (A/R) injury. Ganoderma atrum polysaccharide (PSG-1), the most abundant component isolated from G. atrum, has been shown to possess potent antioxidant activity. The goals of this study were to investigate the effect of PSG-1 against oxidative stress induced by A/R injury and the possible mechanisms in cardiomyocytes. In this work, primary cultures of neonatal rat cardiomyocytes pretreated with PSG-1 were subjected to A/R and subsequently monitored for cell viability by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The levels of intracellular reactive oxygen species (ROS), apoptosis, and mitochondrial membrane potential (Deltapsi(m)) were determined by flow cytometry. Western blot analysis was used to measure the expression of cytochrome c, Bcl-2 family, and manganese superoxide dismutase (MnSOD) proteins, and the activities of caspase-3 and caspase-9 were determined by a colorimetric method. The results showed that PSG-1 protected against cell death caused by A/R injury in cardiomyocytes. PSG-1 reduced the A/R-induced ROS generation, the loss of mitochondrial membrane potential (Deltapsi(m)), and the release of cytochrome c from the mitochondria into cytosol. PSG-1 inhibited the A/R-stimulated activation of caspase-9 and caspase-3 and alteration of Bcl-2 family proteins. Moreover, PSG-1 significantly increased the protein expression of MnSOD in cardiomyocytes. These findings suggest that PSG-1 significantly attenuates A/R-induced oxidative stress and improves cell survival in cardiomyocytes through mitochondrial pathway.

Li WJ ，Nie SP ，Chen Y ，Xie MY ，He M ，Yu Q ，Yan Y ... -  《-》

Capsaicin Protects Cardiomyocytes against Anoxia/Reoxygenation Injury via Preventing Mitochondrial Dysfunction Mediated by SIRT1.

He H ，Zhou Y ，Huang J ，Wu Z ，Liao Z ，Liu D ，Yin D ，He M ... -  《-》

Orientin-induced cardioprotection against reperfusion is associated with attenuation of mitochondrial permeability transition.

In this study, we provide new evidence that orientin from bamboo leaves (Phyllostachys nigra) protect H9c2 cardiomyocytes against ischemia/reperfusion (I/R) injury through the mitochondrial apoptotic pathway. A previous work has identified that orientin could protect myocardium against ischemia/reperfusion injury. Mitochondria are both critical determinants of cardioprotection and crucial targets of cardioprotective signaling. Their role during reperfusion is conspicuously critical because the conditions promote apoptosis through the mitochondrial pathway and necrosis though irreversible damage to mitochondria, which is in association with mitochondrial permeability transition (MPT). After myocardial ischemia, opening of the mPTP is a critical determinant of cell death. The relationship of orientin and mPTP in mediating reperfusion-induced cardiomyocytes injury is still elusive. Here, our results indicate that the protective effect of orientin in H9c2 cells subjected to I/R injury is associated with depression of the mPTP opening, resultant mitochondrial dysfunction, and apoptosis. Further investigation of cellular mechanisms revealed that these effects were associated with inhibition of reactive oxygen species (ROS) generation, repolarization of mitochondrial membrane potential (Δψ(m)), suppression of mitochondrial cytochrome C release, enhancement of the Bcl-2 level, and inhibition of Bax and Smac/DIABLO levels. Furthermore, these beneficial effects of orientin were blocked by the phosphatidylinositol 3-kinase (PI3K) inhibitor wortmannin, and orientin could enhance Akt phosphorylation. In summary, we demonstrate that orientin protects H9c2 cardiomytocytes against I/R-induced apoptosis by modulating the mPTP opening, and this role of orientin may involve the PI3K/Akt signaling pathway.

Lu N ，Sun Y ，Zheng X 《-》