Abstract
Our previous studies demonstrated that high glucose-induced apoptosis in human umbilical vein endothelial cells (HUVECs) is mediated by sequential activation of c-Jun N-terminal kinase (JNK) and caspase, and prevented by exogenous nitric oxide (NO). In this study we further elucidated the roles of the transcriptional factor NF-kappaB, phosphatidylinositol 3'-kinase (PI3K), Akt and endothelial nitric oxide synthase (eNOS) in the apoptosis of HUVECs induced by high glucose. The results showed that high glucose-induced apoptosis was significantly enhanced by PI3K inhibitors (wortmannin and LY294002), NOS inhibitor (NG-nitro-arginine methyl ester) and eNOS antisense oligonucleotide. In contrast, apoptosis was markedly reduced by NF-kappaB inhibitor (pyrrolidine dithiocarbamate, PDTC), NF-kappaB antisense oligonucleotide, NO donor (sodium nitroprusside, SNP), and overexpression of Akt. The high glucose-induced NF-kappaB activation and transient Akt phosphorylation were prevented by the presence of vitamin C. Moreover, high glucose-induced increase in eNOS expression was attenuated by PI3K inhibitors and the negative mutant of PI3K. The activity of JNK induced by high glucose was suppressed by NF-kappaB-specific antisense oligonucleotide. Taken together our results demonstrated that high glucose-induced HUVECs apoptosis is through NF-kappaB-dependent JNK activation and reactive oxygen species (ROS)-dependent Akt dephosphorylation. Activation of the ROS/PI3K/Akt/eNOS signaling pathway in early phase exerts protective effects against the induction of apoptosis by high glucose.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Androstadienes / pharmacology
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Apoptosis*
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Caspase 3
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Caspases / genetics
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Caspases / physiology
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Cell Survival / drug effects
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Cell Survival / physiology
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Cells, Cultured
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Chromones / pharmacology
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Endothelial Cells / cytology
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Endothelial Cells / drug effects
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Endothelial Cells / physiology*
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Female
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Gene Expression Regulation / drug effects
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Gene Expression Regulation / physiology
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Glucose / pharmacology*
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Humans
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JNK Mitogen-Activated Protein Kinases / genetics
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JNK Mitogen-Activated Protein Kinases / physiology*
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Morpholines / pharmacology
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Mutation
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NF-kappa B / antagonists & inhibitors
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NF-kappa B / genetics
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NF-kappa B / metabolism*
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Nitric Oxide / pharmacology
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Nitric Oxide Synthase Type III / genetics
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Nitric Oxide Synthase Type III / physiology*
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Nitroarginine / pharmacology
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Nitroprusside / pharmacology
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Oligonucleotides, Antisense / pharmacology
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Oncogene Protein v-akt / genetics
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Oncogene Protein v-akt / physiology*
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Phosphatidylinositol 3-Kinases / genetics
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Phosphatidylinositol 3-Kinases / physiology*
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Phosphoinositide-3 Kinase Inhibitors
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Phosphorylation
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Proline / analogs & derivatives
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Proline / pharmacology
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Reactive Oxygen Species / metabolism
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Signal Transduction
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Thiocarbamates / pharmacology
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Umbilical Veins / cytology
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Wortmannin
Substances
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Androstadienes
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Chromones
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Morpholines
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NF-kappa B
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Oligonucleotides, Antisense
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Phosphoinositide-3 Kinase Inhibitors
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Reactive Oxygen Species
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Thiocarbamates
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prolinedithiocarbamate
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Nitroprusside
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Nitroarginine
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Nitric Oxide
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2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
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Proline
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NOS3 protein, human
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Nitric Oxide Synthase Type III
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Oncogene Protein v-akt
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JNK Mitogen-Activated Protein Kinases
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CASP3 protein, human
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Caspase 3
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Caspases
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Glucose
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Wortmannin