Phosphocreatine protects endothelial cells from oxidized low-density lipoprotein-induced apoptosis by modulating the PI3K/Akt/eNOS pathway

Abstract

Endothelial apoptosis triggered by oxidized low-density lipoprotein (oxLDL) can accelerate the progression of endothelial dysfunction atherosclerosis. Phosphocreatine (PCr) is a natural compound, which has been used in cardiac disease and cardiopulmonary resuscitation. However, its protective effects on atherosclerosis and its mechanism have not been clarified. In the present study, we investigated the anti-apoptotic effect of phosphocreatine in human umbilical vein endothelial cells (HUVECs) exposed to oxLDL and explored the possible mechanisms. HUVECs were pre-treated with 10-30 mM PCr and then stimulated with oxLDL. Cell morphology, cytotoxicity and apoptosis were evaluated by light microscopy, CCK assay, and flow cytometry respectively. Levels of Bax, Bcl-2, protein expression of protein kinase B (Akt), eNOS and caspase activities were assessed by Western blotting. Reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) were measured with fluorescent probes. Lactate dehydrogenase (LDH), malondialdehyde (MDA), nitric oxide (NO) and superoxide dismutase (SOD) contents were determined by spectrophotometer. Our results showed that PCr dose-dependently prevented oxLDL associated HUVEC cytotoxicity and apoptotic biochemical changes such as loss of MMP, LDH and MDA leakage and loss of SOD, decrease of Bcl-2/Bax protein ratio, activation of caspase-3 and 9, and ROS generation. In addition, the antiapoptotic effect of PCr was partially inhibited by a PI3K inhibitor (LY294002) and also enhanced p-Akt/Akt protein ratio, eNOS activation and NO production. In conclusion, our data show that the inhibition of oxLDL-induced endothelial apoptosis by PCr is due, at least in part to its anti-oxidant activity and its ability to modulate the PI3K/Akt/eNOS signaling pathway.

Pinoresinol Diglucoside Alleviates oxLDL‐Induced Dysfunction in Human Umbilical Vein Endothelial Cells

“…The activation of arginase II by oxLDL was through a LOX-1 dependent RhoA/ROCK pathway [29], which was consistent with the observation that arginase II was dissociated from microtubules by oxLDL [28]. Phosphocreatine antagonized the apoptotic effect of oxLDL by activating PI3K/Akt/eNOS pathway and NO production in HUVECs [30]. Meanwhile, p38MAPK/NF- κ B pathway has been shown to be strongly activated by oxLDL and served as an effective target for intervention [31, 32].…”

SIRT1/AMPK and Akt/eNOS signaling pathways are involved in endothelial protection of total aralosides of Aralia elata (Miq) Seem against high‐fat diet‐induced atherosclerosis in ApoE−/− mice

“…Activated Akt could inhibit the expression of pro‐apoptotic proteins, such as bax, caspase‐9, and caspase‐3 (Hwang et al, ). Moreover, Akt is an important regulator of eNOS activity in vessels (Ahsan et al, ). eNOS regulates the release of NO to promote endothelial cell migration and neovascularization (Fu, Wang, Guo, Xu, & Wu, ).…”

Klotho ameliorates oxidized low density lipoprotein (ox-LDL)-induced oxidative stress via regulating LOX-1 and PI3K/Akt/eNOS pathways

“…The current study suggests that Klotho may rescue ox-LDL-induced inhibition of NO production through the activation of Akt/eNOS pathway in HUVECs. Similarly, some natural compounds also protect endothelial cells from ox-LDL-induced apoptosis by modulating the PI3K/Akt/eNOS pathway [44]. In contrast, many studies indicated that decreased NO is another key mechanism underlying endothelial dysfunction, which is interrelated with decreased expression of the Klotho gene [33, 45–47].…”