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Article: Role of protein kinase C β2 activation in TNF-α-induced human vascular endothelial cell apoptosis

TitleRole of protein kinase C β2 activation in TNF-α-induced human vascular endothelial cell apoptosis
Authors
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
Citation
Canadian Journal Of Physiology And Pharmacology, 2009, v. 87 n. 3, p. 221-229 How to Cite?
Abstract
The 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.
Persistent Identifierhttp://hdl.handle.net/10722/147265
ISSN
2013 Impact Factor: 1.546
ISI Accession Number ID
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).

References

 

Author Affiliations
  1. Hubei General Hospital
  2. The University of Hong Kong
  3. Wuhan University
DC FieldValueLanguage
dc.contributor.authorWang, Fen_US
dc.contributor.authorLiu, HMen_US
dc.contributor.authorIrwin, MGen_US
dc.contributor.authorXia, ZYen_US
dc.contributor.authorHuang, Zen_US
dc.contributor.authorOuyang, Jen_US
dc.contributor.authorXia, Zen_US
dc.date.accessioned2012-05-29T06:01:07Z-
dc.date.available2012-05-29T06:01:07Z-
dc.date.issued2009en_US
dc.identifier.citationCanadian Journal Of Physiology And Pharmacology, 2009, v. 87 n. 3, p. 221-229en_US
dc.identifier.issn0008-4212en_US
dc.identifier.urihttp://hdl.handle.net/10722/147265-
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.en_US
dc.languageengen_US
dc.publisherNRC Research Press. The Journal's web site is located at http://pubs.nrc-cnrc.gc.ca/cgi-bin/rp/rp2_desc_e?cjppen_US
dc.relation.ispartofCanadian Journal of Physiology and Pharmacologyen_US
dc.subject.meshApoptosis - Drug Effectsen_US
dc.subject.meshCell Survival - Drug Effectsen_US
dc.subject.meshCells, Cultureden_US
dc.subject.meshEndothelial Cells - Drug Effects - Physiologyen_US
dc.subject.meshEnzyme Activationen_US
dc.subject.meshGlutathione Peroxidase - Metabolismen_US
dc.subject.meshHumansen_US
dc.subject.meshHydrogen Peroxide - Metabolismen_US
dc.subject.meshMalondialdehyde - Analysisen_US
dc.subject.meshPhthalimides - Pharmacologyen_US
dc.subject.meshProtein Kinase C - Physiologyen_US
dc.subject.meshSuperoxide Dismutase - Metabolismen_US
dc.subject.meshTumor Necrosis Factor-Alpha - Pharmacologyen_US
dc.titleRole of protein kinase C β2 activation in TNF-α-induced human vascular endothelial cell apoptosisen_US
dc.typeArticleen_US
dc.identifier.emailIrwin, MG:mgirwin@hku.hken_US
dc.identifier.emailXia, Z:zyxia@hkucc.hku.hken_US
dc.identifier.authorityIrwin, MG=rp00390en_US
dc.identifier.authorityXia, Z=rp00532en_US
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1139/Y09-004en_US
dc.identifier.pmid19295663en_US
dc.identifier.scopuseid_2-s2.0-65249102747en_US
dc.identifier.hkuros160850-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-65249102747&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume87en_US
dc.identifier.issue3en_US
dc.identifier.spage221en_US
dc.identifier.epage229en_US
dc.identifier.isiWOS:000265605900007-
dc.publisher.placeCanadaen_US

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