Phosphocreatine protects endothelial cells from Methylglyoxal induced oxidative stress and apoptosis via the regulation of PI3K/Akt/eNOS and NF-κB pathway.

Abstract

Methylglyoxal (MGO), an active metabolite of glucose, can cause cellular injury which has an affinity for the progression of diabetes-associated atherosclerosis. Phosphocreatine (PCr) is a well-known high-energy phosphate compound. However, its protective effects and mechanism in the formation of a diabetes-associated atherosclerosis have not been clarified. In the present study, we investigated whether PCr could prevent MGO-induced apoptosis in human umbilical vascular endothelial cells (HUVECs) and explored the possible mechanisms. Cells were pre-treated with PCr and then stimulated with MGO. Cell morphology, cytotoxicity and apoptosis were assessed by light microscopy, MTT assay, and Annexin V-FITC respectively. Apoptotic-related proteins were evaluated by Western blotting. Reactive oxygen species (ROS) generation, intracellular calcium and mitochondrial membrane potential (MMP) were measured with fluorescent probes. Our results showed that PCr dose-dependently prevented MGO associated HUVEC cytotoxicity and suppressed MGO activated ROS generation as well as apoptotic biochemical changes such as lactate dehydrogenase, malondialdehyde leakage, loss of MMP, decreased Bcl-2/Bax protein ratio, levels of caspase-3 and 9. In addition, the antiapoptotic effect of PCr enhanced p-Akt/Akt protein ratio, NO synthase (eNOS) activation, NO production and cGMP levels and also was partially suppressed by a PI3K inhibitor (LY294002). Furthermore, PCr also inhibited MGO-induced transcriptional activity of Nuclear factor kappa B (NFκB). In conclusion, our data described that PCr exerts an antiapoptotic effect in HUVECs exposed to oxidative stress by MGO through the mitochondrial pathway and the modulation of PI3K/Akt/eNOS and NF-κB signaling pathway. Thus, it might be a candidate therapeutic agent for diabetic-associated cardiovascular diseases.

• Authors: Peng Chu, Guozhu Han, Anil Ahsan, Zhengwu Sun, Shumin Liu, Zonghui Zhang, Bin Sun, Yanlin Song, Yuan Lin, Jinyong Peng, Zeyao Tang
• Journal: Vascular pharmacology (Vascul Pharmacol) Vol. 91 Pg. 26-35 (04 2017) ISSN: 1879-3649 [Electronic] United States
• PMID: 27590258 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2016. Published by Elsevier Inc.

Chemical References

• Antioxidants
• Apoptosis Regulatory Proteins
• NF-kappa B
• Reactive Oxygen Species
• Phosphocreatine
• Pyruvaldehyde
• NOS3 protein, human
• Nitric Oxide Synthase Type III
• Phosphatidylinositol 3-Kinase
• Proto-Oncogene Proteins c-akt

Topics

• Antioxidants (pharmacology)
• Apoptosis (drug effects)
• Apoptosis Regulatory Proteins (metabolism)
• Cells, Cultured
• Cytoprotection
• Dose-Response Relationship, Drug
• Human Umbilical Vein Endothelial Cells (drug effects, enzymology, pathology)
• Humans
• Membrane Potential, Mitochondrial (drug effects)
• Mitochondria (drug effects, metabolism, pathology)
• NF-kappa B (metabolism)
• Nitric Oxide Synthase Type III (metabolism)
• Oxidative Stress (drug effects)
• Phosphatidylinositol 3-Kinase (metabolism)
• Phosphocreatine (pharmacology)
• Proto-Oncogene Proteins c-akt (metabolism)
• Pyruvaldehyde (toxicity)
• Reactive Oxygen Species (metabolism)
• Signal Transduction (drug effects)