Nox1-based NADPH oxidase-derived superoxide is required for VSMC activation by advanced glycation end-products

Alejandra San Martin; Rocio Foncea; Francisco R. Laurindo; Roberto Ebensperger; Kathy K. Griendling; Federico Leighton

doi:10.1016/j.freeradbiomed.2007.02.002

Nox1-based NADPH oxidase-derived superoxide is required for VSMC activation by advanced glycation end-products

Alejandra San Martin, Rocio Foncea, Francisco R. Laurindo, Roberto Ebensperger, Kathy K. Griendling, Federico Leighton

Metabolic and Systems Biology (BMBB)

Research output: Contribution to journal › Article › peer-review

89 Scopus citations

Abstract

Vascular diseases are important clinical complications of diabetes. Advanced glycation end-products (AGE) are mediators of vascular dysfunction, but their effects on vascular smooth muscle cell (VSMC) ROS production are unclear. We studied the source and downstream targets of AGE-mediated ROS and reactive nitrogen species production in these cells. Significant increases in superoxide production in AGE-treated VSMC were measured using lucigenin (7650 ± 433 vs 4485 ± 424 LU/10⁶ cells, p < 0.001) or coelenterazine (277,907 ± 71,295 vs 120,456 ± 4140 LU/10⁶ cells, p < 0.05) and confirmed by ESR spectroscopy. These signals were blocked by the flavin-containing oxidase inhibitor diphenylene iodonium (DPI). AGE-stimulated NF-κB activity was abolished by DPI and the superoxide scavenger MnTBAP. AGE differentially regulated VSMC NADPH oxidase catalytic subunits, stimulating the transcription of Nox1 (201 ± 12.7%, p < 0.0001), while having no effect on Nox4. AGE also increased 3-nitrotyrosine formation, which was inhibited by MnTBAP, DPI, or the NOS inhibitor L-NAME. Regarding the source of NO, AGE stimulated inducible nitric oxide synthase mRNA (1 vs 9.7 ± 3.0, p = 0.046), which was abolished by a NF-κB inhibitor, SOD, catalase, or siRNA against Nox1. This study establishes that AGE activate iNOS in VSMC through a ROS-sensitive, NF-κB-dependent mechanism involving ROS generation by a Nox1-based oxidase.

Original language	English (US)
Pages (from-to)	1671-1679
Number of pages	9
Journal	Free Radical Biology and Medicine
Volume	42
Issue number	11
DOIs	https://doi.org/10.1016/j.freeradbiomed.2007.02.002
State	Published - Jun 1 2007

Bibliographical note

Funding Information:
This work was supported by Projects PUC-PBMEC 2001–2003, Fondecyt 1020486, HL075209 and FAPESP 00/12154-2. The authors thank Drs. Ohara Agusto, Sergei Dikalov, and Marcelo Pedro for their technical help with EPR measurements and Dr. Victoria Velarde for her valuable discussions during the performance of these studies.

Keywords

Advanced glycated end products
Diabetes
Free radicals
Inflammation
NADPH oxidase
Nox1
ROS
Superoxide
iNOS

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Publisher link

10.1016/j.freeradbiomed.2007.02.002

Find It

Find It at U of M Libraries

Cite this

@article{7f2255ab30cf4679b547f980a0c50012,

title = "Nox1-based NADPH oxidase-derived superoxide is required for VSMC activation by advanced glycation end-products",

abstract = "Vascular diseases are important clinical complications of diabetes. Advanced glycation end-products (AGE) are mediators of vascular dysfunction, but their effects on vascular smooth muscle cell (VSMC) ROS production are unclear. We studied the source and downstream targets of AGE-mediated ROS and reactive nitrogen species production in these cells. Significant increases in superoxide production in AGE-treated VSMC were measured using lucigenin (7650 ± 433 vs 4485 ± 424 LU/106 cells, p < 0.001) or coelenterazine (277,907 ± 71,295 vs 120,456 ± 4140 LU/106 cells, p < 0.05) and confirmed by ESR spectroscopy. These signals were blocked by the flavin-containing oxidase inhibitor diphenylene iodonium (DPI). AGE-stimulated NF-κB activity was abolished by DPI and the superoxide scavenger MnTBAP. AGE differentially regulated VSMC NADPH oxidase catalytic subunits, stimulating the transcription of Nox1 (201 ± 12.7%, p < 0.0001), while having no effect on Nox4. AGE also increased 3-nitrotyrosine formation, which was inhibited by MnTBAP, DPI, or the NOS inhibitor L-NAME. Regarding the source of NO, AGE stimulated inducible nitric oxide synthase mRNA (1 vs 9.7 ± 3.0, p = 0.046), which was abolished by a NF-κB inhibitor, SOD, catalase, or siRNA against Nox1. This study establishes that AGE activate iNOS in VSMC through a ROS-sensitive, NF-κB-dependent mechanism involving ROS generation by a Nox1-based oxidase.",

keywords = "Advanced glycated end products, Diabetes, Free radicals, Inflammation, NADPH oxidase, Nox1, ROS, Superoxide, iNOS",

author = "{San Martin}, Alejandra and Rocio Foncea and Laurindo, {Francisco R.} and Roberto Ebensperger and Griendling, {Kathy K.} and Federico Leighton",

note = "Funding Information: This work was supported by Projects PUC-PBMEC 2001–2003, Fondecyt 1020486, HL075209 and FAPESP 00/12154-2. The authors thank Drs. Ohara Agusto, Sergei Dikalov, and Marcelo Pedro for their technical help with EPR measurements and Dr. Victoria Velarde for her valuable discussions during the performance of these studies.",

year = "2007",

month = jun,

day = "1",

doi = "10.1016/j.freeradbiomed.2007.02.002",

language = "English (US)",

volume = "42",

pages = "1671--1679",

journal = "Free Radical Biology and Medicine",

issn = "0891-5849",

publisher = "Elsevier Inc.",

number = "11",

}

TY - JOUR

T1 - Nox1-based NADPH oxidase-derived superoxide is required for VSMC activation by advanced glycation end-products

AU - San Martin, Alejandra

AU - Foncea, Rocio

AU - Laurindo, Francisco R.

AU - Ebensperger, Roberto

AU - Griendling, Kathy K.

AU - Leighton, Federico

N1 - Funding Information: This work was supported by Projects PUC-PBMEC 2001–2003, Fondecyt 1020486, HL075209 and FAPESP 00/12154-2. The authors thank Drs. Ohara Agusto, Sergei Dikalov, and Marcelo Pedro for their technical help with EPR measurements and Dr. Victoria Velarde for her valuable discussions during the performance of these studies.

PY - 2007/6/1

Y1 - 2007/6/1

N2 - Vascular diseases are important clinical complications of diabetes. Advanced glycation end-products (AGE) are mediators of vascular dysfunction, but their effects on vascular smooth muscle cell (VSMC) ROS production are unclear. We studied the source and downstream targets of AGE-mediated ROS and reactive nitrogen species production in these cells. Significant increases in superoxide production in AGE-treated VSMC were measured using lucigenin (7650 ± 433 vs 4485 ± 424 LU/106 cells, p < 0.001) or coelenterazine (277,907 ± 71,295 vs 120,456 ± 4140 LU/106 cells, p < 0.05) and confirmed by ESR spectroscopy. These signals were blocked by the flavin-containing oxidase inhibitor diphenylene iodonium (DPI). AGE-stimulated NF-κB activity was abolished by DPI and the superoxide scavenger MnTBAP. AGE differentially regulated VSMC NADPH oxidase catalytic subunits, stimulating the transcription of Nox1 (201 ± 12.7%, p < 0.0001), while having no effect on Nox4. AGE also increased 3-nitrotyrosine formation, which was inhibited by MnTBAP, DPI, or the NOS inhibitor L-NAME. Regarding the source of NO, AGE stimulated inducible nitric oxide synthase mRNA (1 vs 9.7 ± 3.0, p = 0.046), which was abolished by a NF-κB inhibitor, SOD, catalase, or siRNA against Nox1. This study establishes that AGE activate iNOS in VSMC through a ROS-sensitive, NF-κB-dependent mechanism involving ROS generation by a Nox1-based oxidase.

AB - Vascular diseases are important clinical complications of diabetes. Advanced glycation end-products (AGE) are mediators of vascular dysfunction, but their effects on vascular smooth muscle cell (VSMC) ROS production are unclear. We studied the source and downstream targets of AGE-mediated ROS and reactive nitrogen species production in these cells. Significant increases in superoxide production in AGE-treated VSMC were measured using lucigenin (7650 ± 433 vs 4485 ± 424 LU/106 cells, p < 0.001) or coelenterazine (277,907 ± 71,295 vs 120,456 ± 4140 LU/106 cells, p < 0.05) and confirmed by ESR spectroscopy. These signals were blocked by the flavin-containing oxidase inhibitor diphenylene iodonium (DPI). AGE-stimulated NF-κB activity was abolished by DPI and the superoxide scavenger MnTBAP. AGE differentially regulated VSMC NADPH oxidase catalytic subunits, stimulating the transcription of Nox1 (201 ± 12.7%, p < 0.0001), while having no effect on Nox4. AGE also increased 3-nitrotyrosine formation, which was inhibited by MnTBAP, DPI, or the NOS inhibitor L-NAME. Regarding the source of NO, AGE stimulated inducible nitric oxide synthase mRNA (1 vs 9.7 ± 3.0, p = 0.046), which was abolished by a NF-κB inhibitor, SOD, catalase, or siRNA against Nox1. This study establishes that AGE activate iNOS in VSMC through a ROS-sensitive, NF-κB-dependent mechanism involving ROS generation by a Nox1-based oxidase.

KW - Advanced glycated end products

KW - Diabetes

KW - Free radicals

KW - Inflammation

KW - NADPH oxidase

KW - Nox1

KW - ROS

KW - Superoxide

KW - iNOS

UR - http://www.scopus.com/inward/record.url?scp=34247209253&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34247209253&partnerID=8YFLogxK

U2 - 10.1016/j.freeradbiomed.2007.02.002

DO - 10.1016/j.freeradbiomed.2007.02.002

M3 - Article

C2 - 17462535

AN - SCOPUS:34247209253

SN - 0891-5849

VL - 42

SP - 1671

EP - 1679

JO - Free Radical Biology and Medicine

JF - Free Radical Biology and Medicine

IS - 11

ER -

Nox1-based NADPH oxidase-derived superoxide is required for VSMC activation by advanced glycation end-products

Abstract

Bibliographical note

Keywords

UN SDGs

Publisher link

Find It

Other files and links

Fingerprint

Cite this