Article: Role of protein kinase C β2 activation in TNF-α-induced human vascular endothelial cell apoptosis
| Title | Role of protein kinase C β2 activation in TNF-α-induced human vascular endothelial cell apoptosis | ||||||
|---|---|---|---|---|---|---|---|
| Authors | Wang, F1 Liu, HM1 2 Irwin, MG2 Xia, ZY1 Huang, Z2 Ouyang, J3 Xia, Z1 2 | ||||||
| Issue Date | 2009 | ||||||
| Publisher | NRC 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?] DOI: http://dx.doi.org/10.1139/Y09-004 | ||||||
| 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. | ||||||
| ISSN | 0008-4212 2011 Impact Factor: 1.953 2011 SCImago Journal Rankings: 0.149 | ||||||
| DOI | http://dx.doi.org/10.1139/Y09-004 | ||||||
| ISI Accession Number ID | WOS:000265605900007
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 | References in Scopus |
| dc.contributor.author | Wang, F | ||||||
|---|---|---|---|---|---|---|---|
| dc.contributor.author | Liu, HM | ||||||
| dc.contributor.author | Irwin, MG | ||||||
| dc.contributor.author | Xia, ZY | ||||||
| dc.contributor.author | Huang, Z | ||||||
| dc.contributor.author | Ouyang, J | ||||||
| dc.contributor.author | Xia, Z | ||||||
| dc.date.accessioned | 2012-05-29T06:01:07Z | ||||||
| dc.date.available | 2012-05-29T06:01:07Z | ||||||
| dc.date.issued | 2009 | ||||||
| dc.description.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. | ||||||
| dc.description.nature | Link_to_subscribed_fulltext | ||||||
| dc.identifier.citation | Canadian 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.doi | http://dx.doi.org/10.1139/Y09-004 | ||||||
| dc.identifier.epage | 229 | ||||||
| dc.identifier.hkuros | 160850 | ||||||
| dc.identifier.isi | WOS:000265605900007
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.issn | 0008-4212 2011 Impact Factor: 1.953 2011 SCImago Journal Rankings: 0.149 | ||||||
| dc.identifier.issue | 3 | ||||||
| dc.identifier.pmid | 19295663 | ||||||
| dc.identifier.scopus | eid_2-s2.0-65249102747 | ||||||
| dc.identifier.spage | 221 | ||||||
| dc.identifier.uri | http://hdl.handle.net/10722/147265 | ||||||
| dc.identifier.volume | 87 | ||||||
| dc.language | eng | ||||||
| dc.publisher | NRC 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.place | Canada | ||||||
| dc.relation.ispartof | Canadian Journal of Physiology and Pharmacology | ||||||
| dc.relation.references | References in Scopus | ||||||
| dc.subject.mesh | Apoptosis - Drug Effects | ||||||
| dc.subject.mesh | Cell Survival - Drug Effects | ||||||
| dc.subject.mesh | Cells, Cultured | ||||||
| dc.subject.mesh | Endothelial Cells - Drug Effects - Physiology | ||||||
| dc.subject.mesh | Enzyme Activation | ||||||
| dc.subject.mesh | Glutathione Peroxidase - Metabolism | ||||||
| dc.subject.mesh | Humans | ||||||
| dc.subject.mesh | Hydrogen Peroxide - Metabolism | ||||||
| dc.subject.mesh | Malondialdehyde - Analysis | ||||||
| dc.subject.mesh | Phthalimides - Pharmacology | ||||||
| dc.subject.mesh | Protein Kinase C - Physiology | ||||||
| dc.subject.mesh | Superoxide Dismutase - Metabolism | ||||||
| dc.subject.mesh | Tumor Necrosis Factor-Alpha - Pharmacology | ||||||
| dc.title | Role of protein kinase C β2 activation in TNF-α-induced human vascular endothelial cell apoptosis | ||||||
| dc.type | Article |
Author Affiliations
- Hubei General Hospital
- The University of Hong Kong
- Wuhan University