title: 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: Pharmacology Department, Dalian Medical University, Dalian, China.
authors: Peng
authors: P
authors: Chu
authors: Pharmacology Department, Dalian Medical University, Dalian, China.
authors: Guozhu
authors: G
authors: Han
authors: Pharmacology Department, Dalian Medical University, Dalian, China.
authors: Anil
authors: A
authors: Ahsan
authors: Pharmacology Department, Dalian Medical University, Dalian, China.
authors: Zhengwu
authors: Z
authors: Sun
authors: Pharmacology Department, Dalian Medical University, Dalian, China.
authors: Shumin
authors: S
authors: Liu
authors: Pharmacology Department, Dalian Medical University, Dalian, China.
authors: Zonghui
authors: Z
authors: Zhang
authors: Pharmacology Department, Dalian Medical University, Dalian, China.
authors: Bin
authors: B
authors: Sun
authors: Pharmacology Department, Dalian Medical University, Dalian, China.
authors: Yanlin
authors: Y
authors: Song
authors: Pharmacology Department, Dalian Medical University, Dalian, China.
authors: Yuan
authors: Y
authors: Lin
authors: Pharmacology Department, Dalian Medical University, Dalian, China.
authors: Jinyong
authors: J
authors: Peng
authors: Pharmacology Department, Dalian Medical University, Dalian, China. Electronic address: tangzeyao@aliyun.com.
authors: Zeyao
authors: Z
authors: Tang
mesh_terms: D000975:Antioxidants / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D002478:Cells, Cultured; D019610:Cytoprotection; D004305:Dose-Response Relationship, Drug; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D016328:NF-kappa B / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism*; D018384:Oxidative Stress / Q000187:drug effects*; D058539:Phosphatidylinositol 3-Kinase / Q000378:metabolism*; D010725:Phosphocreatine / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D011765:Pyruvaldehyde / Q000633:toxicity*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D000975:Antioxidants; D051017:Apoptosis Regulatory Proteins; D016328:NF-kappa B; D017382:Reactive Oxygen Species; D010725:Phosphocreatine; D011765:Pyruvaldehyde; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D058539:Phosphatidylinositol 3-Kinase; D051057:Proto-Oncogene Proteins c-akt
keywords: Apoptosis; Endothelial cells; Methylglyoxal; Oxidative stress; Phosphocreatine