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Article: Inhibition of Protein Kinase C β2 Prevents Tumor Necrosis Factor-α-Induced Apoptosis and Oxidative Stress in Endothelial Cells: The Role of NADPH Oxidase Subunits

TitleInhibition of Protein Kinase C β2 Prevents Tumor Necrosis Factor-α-Induced Apoptosis and Oxidative Stress in Endothelial Cells: The Role of NADPH Oxidase Subunits
Authors
KeywordsApoptosis
Human Umbilical Vein Endothelial Cells
Nadph Oxidase
Protein Kinase C Β 2
Reactive Oxygen Species
Issue Date2012
PublisherS Karger AG. The Journal's web site is located at http://www.karger.com/JVR
Citation
Journal Of Vascular Research, 2012, v. 49 n. 2, p. 144-159 How to Cite?
AbstractWe investigate the cell signal transduction pathway protein kinase C (PKC) and the role of NADPH subunits in the process of TNF-α-induced endothelial apoptosis. Human umbilical vein endothelial cells (HUVEC) were treated with one of these: 1 mM PKC β 2 inhibitor CGP53353, 10 mM PKC δ inhibitor rottlerin, combination CGP53353 with rottlerin, 3 ×10 -4M NADPH oxidase inhibitor apocynin, 5 × 10 -6M NADPH oxidase peptide inhibitor gp91ds-tat. The apoptosis process was assessed by Hoechst 33342 stain, flow cytometry and Western blot analysis, while intracellular reactive oxygen species (ROS) production was detected by 2,7'-dichlorodihydrofluorescein diacetate (DCFH-DA). The NADPH oxidase subunit gene and protein expression were assessed by quantitative real-time PCR and Western blot analysis, respectively. TNF-α significantly induced HUVEC apoptosis and ROS production, accompanying with dramatic upregulation of NADPH oxidase subunits: NOX2/gp91 phox, NOX4, p47 phox and p67 phox, whereas these enhancements were abolished by the treatment with PKC inhibitors. High TNF-α level exposure induces HUVEC apoptosis, as well as a ROS generation increase via the PKC β 2-dependent activation of NADPH oxidase. Although the PKC δ pathway may enhance TNF-α-induced HUVEC apoptosis, it does not involve the ROS pathway. Upregulation of expression of NADPH subunits is important in this process, which leads to a new target in antioxidative therapy for vascular disease prevention. Copyright © 2012 S. Karger AG.
Persistent Identifierhttp://hdl.handle.net/10722/147287
ISSN
2014 Impact Factor: 2.901
2014 SCImago Journal Rankings: 1.097
ISI Accession Number ID
Funding AgencyGrant Number
National Nature Science Foundation of China30770899
Nature Science Foundation of Guangdong province of China815100890100072
Funding Information:

We are grateful to Professor Rui Zhang (Department of Maternity, Sun Yat-Sen Memorial Hospital, Sun Yet-Sen University) for her helpful assistance, and to Dr. Huimin Liu (Anesthesiology Research Laboratory, Renmin Hospital, Wuhan University) for scientific advice. This work was supported in part by grants from the National Nature Science Foundation of China (No. 30770899 to R.N.) and the Nature Science Foundation of Guangdong province of China (No. 815100890100072 to R.N.).

References

 

DC FieldValueLanguage
dc.contributor.authorDeng, Ben_US
dc.contributor.authorXie, Sen_US
dc.contributor.authorWang, Jen_US
dc.contributor.authorXia, Zen_US
dc.contributor.authorNie, Ren_US
dc.date.accessioned2012-05-29T06:01:15Z-
dc.date.available2012-05-29T06:01:15Z-
dc.date.issued2012en_US
dc.identifier.citationJournal Of Vascular Research, 2012, v. 49 n. 2, p. 144-159en_US
dc.identifier.issn1018-1172en_US
dc.identifier.urihttp://hdl.handle.net/10722/147287-
dc.description.abstractWe investigate the cell signal transduction pathway protein kinase C (PKC) and the role of NADPH subunits in the process of TNF-α-induced endothelial apoptosis. Human umbilical vein endothelial cells (HUVEC) were treated with one of these: 1 mM PKC β 2 inhibitor CGP53353, 10 mM PKC δ inhibitor rottlerin, combination CGP53353 with rottlerin, 3 ×10 -4M NADPH oxidase inhibitor apocynin, 5 × 10 -6M NADPH oxidase peptide inhibitor gp91ds-tat. The apoptosis process was assessed by Hoechst 33342 stain, flow cytometry and Western blot analysis, while intracellular reactive oxygen species (ROS) production was detected by 2,7'-dichlorodihydrofluorescein diacetate (DCFH-DA). The NADPH oxidase subunit gene and protein expression were assessed by quantitative real-time PCR and Western blot analysis, respectively. TNF-α significantly induced HUVEC apoptosis and ROS production, accompanying with dramatic upregulation of NADPH oxidase subunits: NOX2/gp91 phox, NOX4, p47 phox and p67 phox, whereas these enhancements were abolished by the treatment with PKC inhibitors. High TNF-α level exposure induces HUVEC apoptosis, as well as a ROS generation increase via the PKC β 2-dependent activation of NADPH oxidase. Although the PKC δ pathway may enhance TNF-α-induced HUVEC apoptosis, it does not involve the ROS pathway. Upregulation of expression of NADPH subunits is important in this process, which leads to a new target in antioxidative therapy for vascular disease prevention. Copyright © 2012 S. Karger AG.en_US
dc.languageengen_US
dc.publisherS Karger AG. The Journal's web site is located at http://www.karger.com/JVRen_US
dc.relation.ispartofJournal of Vascular Researchen_US
dc.subjectApoptosisen_US
dc.subjectHuman Umbilical Vein Endothelial Cellsen_US
dc.subjectNadph Oxidaseen_US
dc.subjectProtein Kinase C Β 2en_US
dc.subjectReactive Oxygen Speciesen_US
dc.titleInhibition of Protein Kinase C β2 Prevents Tumor Necrosis Factor-α-Induced Apoptosis and Oxidative Stress in Endothelial Cells: The Role of NADPH Oxidase Subunitsen_US
dc.typeArticleen_US
dc.identifier.emailXia, Z:zyxia@hkucc.hku.hken_US
dc.identifier.authorityXia, Z=rp00532en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.description.naturelink_to_OA_fulltext-
dc.identifier.doi10.1159/000332337en_US
dc.identifier.pmid22261918-
dc.identifier.scopuseid_2-s2.0-84862811283-
dc.identifier.hkuros228601-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-84855883897&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume49en_US
dc.identifier.issue2en_US
dc.identifier.spage144en_US
dc.identifier.epage159en_US
dc.identifier.isiWOS:000303152300006-
dc.publisher.placeSwitzerlanden_US

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