Article: Inhibition of protein kinase C β 2 prevents tumor necrosis factor-α-induced apoptosis and oxidative stress in endothelial cells: The role of NADPH oxidase subunits
| Title | Inhibition of protein kinase C β 2 prevents tumor necrosis factor-α-induced apoptosis and oxidative stress in endothelial cells: The role of NADPH oxidase subunits | ||||||
|---|---|---|---|---|---|---|---|
| Authors | Deng, B2 Xie, S2 Wang, J2 Xia, Z1 Nie, R2 | ||||||
| Keywords | Apoptosis Human Umbilical Vein Endothelial Cells Nadph Oxidase Protein Kinase C Β 2 Reactive Oxygen Species | ||||||
| Issue Date | 2012 | ||||||
| Publisher | S 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?] DOI: http://dx.doi.org/10.1159/000332337 | ||||||
| Abstract | We 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. | ||||||
| ISSN | 1018-1172 2011 Impact Factor: 2.651 2011 SCImago Journal Rankings: 0.218 | ||||||
| DOI | http://dx.doi.org/10.1159/000332337 | ||||||
| ISI Accession Number ID | WOS:000303152300006
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 | References in Scopus |
| dc.contributor.author | Deng, B | ||||||
|---|---|---|---|---|---|---|---|
| dc.contributor.author | Xie, S | ||||||
| dc.contributor.author | Wang, J | ||||||
| dc.contributor.author | Xia, Z | ||||||
| dc.contributor.author | Nie, R | ||||||
| dc.date.accessioned | 2012-05-29T06:01:15Z | ||||||
| dc.date.available | 2012-05-29T06:01:15Z | ||||||
| dc.date.issued | 2012 | ||||||
| dc.description.abstract | We 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. | ||||||
| dc.description.nature | Link_to_subscribed_fulltext | ||||||
| dc.identifier.citation | Journal Of Vascular Research, 2012, v. 49 n. 2, p. 144-159 [How to Cite?] DOI: http://dx.doi.org/10.1159/000332337 | ||||||
| dc.identifier.doi | http://dx.doi.org/10.1159/000332337 | ||||||
| dc.identifier.epage | 159 | ||||||
| dc.identifier.isi | WOS:000303152300006
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.). | ||||||
| dc.identifier.issn | 1018-1172 2011 Impact Factor: 2.651 2011 SCImago Journal Rankings: 0.218 | ||||||
| dc.identifier.issue | 2 | ||||||
| dc.identifier.pmid | 22261918 | ||||||
| dc.identifier.scopus | eid_2-s2.0-84862811283 | ||||||
| dc.identifier.spage | 144 | ||||||
| dc.identifier.uri | http://hdl.handle.net/10722/147287 | ||||||
| dc.identifier.volume | 49 | ||||||
| dc.language | eng | ||||||
| dc.publisher | S Karger AG. The Journal's web site is located at http://www.karger.com/JVR | ||||||
| dc.publisher.place | Switzerland | ||||||
| dc.relation.ispartof | Journal of Vascular Research | ||||||
| dc.relation.references | References in Scopus | ||||||
| dc.subject | Apoptosis | ||||||
| dc.subject | Human Umbilical Vein Endothelial Cells | ||||||
| dc.subject | Nadph Oxidase | ||||||
| dc.subject | Protein Kinase C Β 2 | ||||||
| dc.subject | Reactive Oxygen Species | ||||||
| dc.title | Inhibition of protein kinase C β 2 prevents tumor necrosis factor-α-induced apoptosis and oxidative stress in endothelial cells: The role of NADPH oxidase subunits | ||||||
| dc.type | Article |
Author Affiliations
- Wuhan University
- Sun Yat-Sen University