Role of NADPH oxidase 4 in lipopolysaccharide-induced proinflammatory responses by human aortic endothelial cells

Hye Sun Park; Jung Nyeo Chun; Hye Young Jung; Chulhee Choi; Yun Soo Bae

doi:10.1016/j.cardiores.2006.09.012

Role of NADPH oxidase 4 in lipopolysaccharide-induced proinflammatory responses by human aortic endothelial cells

Hye Sun Park, Jung Nyeo Chun, Hye Young Jung, Chulhee Choi, Yun Soo Bae

Department of Life Science

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

194 Scopus citations

Abstract

Objective: We investigated the role of NADPH oxidase 4 (Nox4) on lipopolysaccharide (LPS)-induced proinflammatory responses by human aortic endothelial cells (HAECs). Methods and results: Yeast two-hybrid and glutathione-S-transferase pull-down assays indicated that the cytosolic Toll/IL-1R region of Toll-like receptor 4 (TLR4) (amino acids 739-769) is the responsible domain for interaction with the COOH terminal of Nox4 (amino acids 451-530). Consistently, overexpression of the COOH-terminal region of Nox4 inhibited nuclear factor-κB activation in response to LPS. Downregulation of Nox4 by transfection of siRNA specific to Nox4 in HAECs resulted in a failure to induce reactive oxygen species (ROS) generation and subsequent expression of intercellular adhesion molecule-1 (ICAM-1) and chemokines such as IL-8 and monocyte chemoattractant protein-1 (MCP-1) in response to LPS. Furthermore, transient transfection of endothelial cells with Nox4 siRNA led to a decrease in migration and adhesion of monocytes in response to LPS by 36% and 52%, respectively. Conclusions: Nox4 plays a central role in LPS-induced proinflammatory responses by endothelial cells in an ROS-dependent manner.

Original language	English
Pages (from-to)	447-455
Number of pages	9
Journal	Cardiovascular Research
Volume	72
Issue number	3
DOIs	https://doi.org/10.1016/j.cardiores.2006.09.012
State	Published - 1 Dec 2006

Bibliographical note

Funding Information:
This work is supported by the NCRC program of MOST/ KOSEF (Grant # R15-2006-020-00000-0) through the Center for Cell Signaling & Drug Discovery Research at Ewha Womans University, by 21st century Frontier Functional Proteomics Project (FPR05C2-510) from the Ministry of Science and Technology, by a grant of the Korea Health 21 R&D Project (grant number A06-00043579), Ministry of Health and Welfare and by Seoul R&BD program (grant number 10527). We thank Dr. Jaesang Kim for critical reading.

Keywords

Atherosclerosis
Endothelial cells
Lipopolysaccharide
NADPH oxidase 4
Reactive oxygen species
Toll-like receptor 4

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10.1016/j.cardiores.2006.09.012

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title = "Role of NADPH oxidase 4 in lipopolysaccharide-induced proinflammatory responses by human aortic endothelial cells",

abstract = "Objective: We investigated the role of NADPH oxidase 4 (Nox4) on lipopolysaccharide (LPS)-induced proinflammatory responses by human aortic endothelial cells (HAECs). Methods and results: Yeast two-hybrid and glutathione-S-transferase pull-down assays indicated that the cytosolic Toll/IL-1R region of Toll-like receptor 4 (TLR4) (amino acids 739-769) is the responsible domain for interaction with the COOH terminal of Nox4 (amino acids 451-530). Consistently, overexpression of the COOH-terminal region of Nox4 inhibited nuclear factor-κB activation in response to LPS. Downregulation of Nox4 by transfection of siRNA specific to Nox4 in HAECs resulted in a failure to induce reactive oxygen species (ROS) generation and subsequent expression of intercellular adhesion molecule-1 (ICAM-1) and chemokines such as IL-8 and monocyte chemoattractant protein-1 (MCP-1) in response to LPS. Furthermore, transient transfection of endothelial cells with Nox4 siRNA led to a decrease in migration and adhesion of monocytes in response to LPS by 36% and 52%, respectively. Conclusions: Nox4 plays a central role in LPS-induced proinflammatory responses by endothelial cells in an ROS-dependent manner.",

keywords = "Atherosclerosis, Endothelial cells, Lipopolysaccharide, NADPH oxidase 4, Reactive oxygen species, Toll-like receptor 4",

author = "Park, {Hye Sun} and Chun, {Jung Nyeo} and Jung, {Hye Young} and Chulhee Choi and Bae, {Yun Soo}",

note = "Funding Information: This work is supported by the NCRC program of MOST/ KOSEF (Grant # R15-2006-020-00000-0) through the Center for Cell Signaling & Drug Discovery Research at Ewha Womans University, by 21st century Frontier Functional Proteomics Project (FPR05C2-510) from the Ministry of Science and Technology, by a grant of the Korea Health 21 R&D Project (grant number A06-00043579), Ministry of Health and Welfare and by Seoul R&BD program (grant number 10527). We thank Dr. Jaesang Kim for critical reading.",

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doi = "10.1016/j.cardiores.2006.09.012",

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AU - Park, Hye Sun

AU - Chun, Jung Nyeo

AU - Jung, Hye Young

AU - Choi, Chulhee

AU - Bae, Yun Soo

N1 - Funding Information: This work is supported by the NCRC program of MOST/ KOSEF (Grant # R15-2006-020-00000-0) through the Center for Cell Signaling & Drug Discovery Research at Ewha Womans University, by 21st century Frontier Functional Proteomics Project (FPR05C2-510) from the Ministry of Science and Technology, by a grant of the Korea Health 21 R&D Project (grant number A06-00043579), Ministry of Health and Welfare and by Seoul R&BD program (grant number 10527). We thank Dr. Jaesang Kim for critical reading.

PY - 2006/12/1

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N2 - Objective: We investigated the role of NADPH oxidase 4 (Nox4) on lipopolysaccharide (LPS)-induced proinflammatory responses by human aortic endothelial cells (HAECs). Methods and results: Yeast two-hybrid and glutathione-S-transferase pull-down assays indicated that the cytosolic Toll/IL-1R region of Toll-like receptor 4 (TLR4) (amino acids 739-769) is the responsible domain for interaction with the COOH terminal of Nox4 (amino acids 451-530). Consistently, overexpression of the COOH-terminal region of Nox4 inhibited nuclear factor-κB activation in response to LPS. Downregulation of Nox4 by transfection of siRNA specific to Nox4 in HAECs resulted in a failure to induce reactive oxygen species (ROS) generation and subsequent expression of intercellular adhesion molecule-1 (ICAM-1) and chemokines such as IL-8 and monocyte chemoattractant protein-1 (MCP-1) in response to LPS. Furthermore, transient transfection of endothelial cells with Nox4 siRNA led to a decrease in migration and adhesion of monocytes in response to LPS by 36% and 52%, respectively. Conclusions: Nox4 plays a central role in LPS-induced proinflammatory responses by endothelial cells in an ROS-dependent manner.

AB - Objective: We investigated the role of NADPH oxidase 4 (Nox4) on lipopolysaccharide (LPS)-induced proinflammatory responses by human aortic endothelial cells (HAECs). Methods and results: Yeast two-hybrid and glutathione-S-transferase pull-down assays indicated that the cytosolic Toll/IL-1R region of Toll-like receptor 4 (TLR4) (amino acids 739-769) is the responsible domain for interaction with the COOH terminal of Nox4 (amino acids 451-530). Consistently, overexpression of the COOH-terminal region of Nox4 inhibited nuclear factor-κB activation in response to LPS. Downregulation of Nox4 by transfection of siRNA specific to Nox4 in HAECs resulted in a failure to induce reactive oxygen species (ROS) generation and subsequent expression of intercellular adhesion molecule-1 (ICAM-1) and chemokines such as IL-8 and monocyte chemoattractant protein-1 (MCP-1) in response to LPS. Furthermore, transient transfection of endothelial cells with Nox4 siRNA led to a decrease in migration and adhesion of monocytes in response to LPS by 36% and 52%, respectively. Conclusions: Nox4 plays a central role in LPS-induced proinflammatory responses by endothelial cells in an ROS-dependent manner.

KW - Atherosclerosis

KW - Endothelial cells

KW - Lipopolysaccharide

KW - NADPH oxidase 4

KW - Reactive oxygen species

KW - Toll-like receptor 4

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EP - 455

JO - Cardiovascular Research

JF - Cardiovascular Research

IS - 3

ER -

Role of NADPH oxidase 4 in lipopolysaccharide-induced proinflammatory responses by human aortic endothelial cells

Abstract

Bibliographical note

Keywords

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