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Article: Role of protein kinase C β2 activation in TNF-α-induced human vascular endothelial cell apoptosis
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TitleRole of protein kinase C β2 activation in TNF-α-induced human vascular endothelial cell apoptosis
 
AuthorsWang, F1
Liu, HM1 2
Irwin, MG2
Xia, ZY1
Huang, Z2
Ouyang, J3
Xia, Z1 2
 
Issue Date2009
 
PublisherNRC Research Press. The Journal's web site is located at http://pubs.nrc-cnrc.gc.ca/cgi-bin/rp/rp2_desc_e?cjpp
 
CitationCanadian Journal Of Physiology And Pharmacology, 2009, v. 87 n. 3, p. 221-229 [How to Cite?]
DOI: http://dx.doi.org/10.1139/Y09-004
 
AbstractThe circulatory inflammatory cytokine tumor necrosis factor alpha (TNF-α) is increased in pathologic conditions that initiate or exacerbate vascular endothelial injury, such as diabetes. Protein kinase C (PKC) has been shown to play a critical role in TNF-α-induced human endothelial cell apoptosis. However, the relative roles played by specific isoforms of PKC in TNF-α-induced human endothelial cell apoptosis have not been addressed. We investigated the effects of a selective PKCβ2 inhibitor (CGP53353) on TNF-α-induced apoptosis in human vascular endothelial cells (cell line ECV304) and on the production of reactive oxygen species and nitric oxide, and compared its effects with rottlerin, a reagent that has been shown to reduce PKCδ protein levels. Cultured human vascular endothelial cells (ECV304) were treated for 24 h with one of 4 regimes: 40 ng/mL TNF-α alone (TNF-α), TNF-α with 10 μmol/L rottlerin (T+rottlerin), TNF-α with 1 μmol/L CGP53353 (T+CGP), or untreated (control). Cell viability was measured by MTT assay, and cell apoptosis was assessed by flow cytometry. TNF-a-induced endothelial cell apoptosis was associated with dramatic increases in production of intracellular hydrogen peroxide (approximately 20 times greater than control) and superoxide (approximately 16 times greater than control), as measured by dichlorofluorescein and dihydroethidium fluorescent staining, respectively. This increase was accompanied by reduced activity of superoxide dismutase and glutathione peroxidase and, subsequently, an increase in the lipid peroxidation product malondialdehyde. CGP53353, but not rottlerin, abolished or attenuated all these changes. We conclude that PKCβ 2plays a major role in TNF-α-induced human vascular endothelial cell apoptosis.
 
ISSN0008-4212
2012 Impact Factor: 1.556
2012 SCImago Journal Rankings: 0.550
 
DOIhttp://dx.doi.org/10.1139/Y09-004
 
ISI Accession Number IDWOS:000265605900007
Funding AgencyGrant Number
National Natural Science Foundation of China (NSFC)30872447
30672033
University of Hong KongURC 200801159006
Funding Information:

This study was supported in part by grants 30872447 and 30672033 from the National Natural Science Foundation of China (NSFC), and in part by the Seeding Funding Programme for Basic Research from the University of Hong Kong (grant URC 200801159006).

 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorWang, F
 
dc.contributor.authorLiu, HM
 
dc.contributor.authorIrwin, MG
 
dc.contributor.authorXia, ZY
 
dc.contributor.authorHuang, Z
 
dc.contributor.authorOuyang, J
 
dc.contributor.authorXia, Z
 
dc.date.accessioned2012-05-29T06:01:07Z
 
dc.date.available2012-05-29T06:01:07Z
 
dc.date.issued2009
 
dc.description.abstractThe circulatory inflammatory cytokine tumor necrosis factor alpha (TNF-α) is increased in pathologic conditions that initiate or exacerbate vascular endothelial injury, such as diabetes. Protein kinase C (PKC) has been shown to play a critical role in TNF-α-induced human endothelial cell apoptosis. However, the relative roles played by specific isoforms of PKC in TNF-α-induced human endothelial cell apoptosis have not been addressed. We investigated the effects of a selective PKCβ2 inhibitor (CGP53353) on TNF-α-induced apoptosis in human vascular endothelial cells (cell line ECV304) and on the production of reactive oxygen species and nitric oxide, and compared its effects with rottlerin, a reagent that has been shown to reduce PKCδ protein levels. Cultured human vascular endothelial cells (ECV304) were treated for 24 h with one of 4 regimes: 40 ng/mL TNF-α alone (TNF-α), TNF-α with 10 μmol/L rottlerin (T+rottlerin), TNF-α with 1 μmol/L CGP53353 (T+CGP), or untreated (control). Cell viability was measured by MTT assay, and cell apoptosis was assessed by flow cytometry. TNF-a-induced endothelial cell apoptosis was associated with dramatic increases in production of intracellular hydrogen peroxide (approximately 20 times greater than control) and superoxide (approximately 16 times greater than control), as measured by dichlorofluorescein and dihydroethidium fluorescent staining, respectively. This increase was accompanied by reduced activity of superoxide dismutase and glutathione peroxidase and, subsequently, an increase in the lipid peroxidation product malondialdehyde. CGP53353, but not rottlerin, abolished or attenuated all these changes. We conclude that PKCβ 2plays a major role in TNF-α-induced human vascular endothelial cell apoptosis.
 
dc.description.natureLink_to_subscribed_fulltext
 
dc.identifier.citationCanadian Journal Of Physiology And Pharmacology, 2009, v. 87 n. 3, p. 221-229 [How to Cite?]
DOI: http://dx.doi.org/10.1139/Y09-004
 
dc.identifier.doihttp://dx.doi.org/10.1139/Y09-004
 
dc.identifier.epage229
 
dc.identifier.hkuros160850
 
dc.identifier.isiWOS:000265605900007
Funding AgencyGrant Number
National Natural Science Foundation of China (NSFC)30872447
30672033
University of Hong KongURC 200801159006
Funding Information:

This study was supported in part by grants 30872447 and 30672033 from the National Natural Science Foundation of China (NSFC), and in part by the Seeding Funding Programme for Basic Research from the University of Hong Kong (grant URC 200801159006).

 
dc.identifier.issn0008-4212
2012 Impact Factor: 1.556
2012 SCImago Journal Rankings: 0.550
 
dc.identifier.issue3
 
dc.identifier.pmid19295663
 
dc.identifier.scopuseid_2-s2.0-65249102747
 
dc.identifier.spage221
 
dc.identifier.urihttp://hdl.handle.net/10722/147265
 
dc.identifier.volume87
 
dc.languageeng
 
dc.publisherNRC Research Press. The Journal's web site is located at http://pubs.nrc-cnrc.gc.ca/cgi-bin/rp/rp2_desc_e?cjpp
 
dc.publisher.placeCanada
 
dc.relation.ispartofCanadian Journal of Physiology and Pharmacology
 
dc.relation.referencesReferences in Scopus
 
dc.subject.meshApoptosis - Drug Effects
 
dc.subject.meshCell Survival - Drug Effects
 
dc.subject.meshCells, Cultured
 
dc.subject.meshEndothelial Cells - Drug Effects - Physiology
 
dc.subject.meshEnzyme Activation
 
dc.subject.meshGlutathione Peroxidase - Metabolism
 
dc.subject.meshHumans
 
dc.subject.meshHydrogen Peroxide - Metabolism
 
dc.subject.meshMalondialdehyde - Analysis
 
dc.subject.meshPhthalimides - Pharmacology
 
dc.subject.meshProtein Kinase C - Physiology
 
dc.subject.meshSuperoxide Dismutase - Metabolism
 
dc.subject.meshTumor Necrosis Factor-Alpha - Pharmacology
 
dc.titleRole of protein kinase C β2 activation in TNF-α-induced human vascular endothelial cell apoptosis
 
dc.typeArticle
 
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<contributor.author>Xia, ZY</contributor.author>
<contributor.author>Huang, Z</contributor.author>
<contributor.author>Ouyang, J</contributor.author>
<contributor.author>Xia, Z</contributor.author>
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<subject.mesh>Enzyme Activation</subject.mesh>
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<subject.mesh>Hydrogen Peroxide - Metabolism</subject.mesh>
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Author Affiliations
  1. Hubei General Hospital
  2. The University of Hong Kong
  3. Wuhan University