Chuntao Yang" /> and Jianqiang Feng *

" /> Chuntao Yang, Hongzhong Ling, Meifen Zhang, Zhanli Yang, Xiuyu Wang, Fanqin Zeng, Chuhuai Wang, and Jianqiang Feng*

" /> Chuntao Yang, Hongzhong Ling, Meifen Zhang, Zhanli Yang, Xiuyu Wang, Fanqin Zeng, Chuhuai Wang, and Jianqiang Feng*

. Mol. Cells 2011;31:531-8. https://doi.org/10.1007/s10059-011-1025-3">
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Mol. Cells 2011; 31(6): 531-538

Published online April 20, 2011

https://doi.org/10.1007/s10059-011-1025-3

© The Korean Society for Molecular and Cellular Biology

Oxidative Stress Mediates Chemical Hypoxia-Induced Injury and Inflammation by Activating NF-ĸb-COX-2 Pathway in HaCaT Cells

Chuntao Yang, Hongzhong Ling1, Meifen Zhang2, Zhanli Yang, Xiuyu Wang, Fanqin Zeng3, Chuhuai Wang4, and Jianqiang Feng*

Department of Physiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangdong, People’s Republic of China, 1Department of Dermatology, The First Affiliated Hospital, Sun Yat-sen University, Guangdong, People’s Republic of China, 2School of Nursing, Sun Yat-sen University, Guangdong, People’s Republic of China, 3Department of Dermatology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangdong, People’s Republic of China, 4Department of Rehabilitation, The First Affiliated Hospital, Sun Yat-sen University, Guangdong, People’s Republic of China

Correspondence to : *Correspondence: fengjq-sums@163.com

Received: February 14, 2011; Revised: March 21, 2011; Accepted: March 22, 2011

Abstract

Hypoxia of skin is an important physiopathological process in many diseases, such as pressure ulcer, diabet-ic ulcer, and varicose ulcer. Although cellular injury and inflammation have been involved in hypoxia-induced dermatic injury, the underlying mechanisms remain largely unknown. This study was conducted to investigate the effects of cobalt chloride (CoCl2), a hypoxia-mimicking agent, on human skin keratinocytes (HaCaT cells) and to explore the possible molecular mechanisms. Exposure of HaCaT cells to CoCl2 reduced cell viability and caused overproduction of reactive oxygen species (ROS) and oversecretion of interleukin-6 (IL-6) and interleukin-8 (IL-8). Importantly, CoCl2 exposure elicited overexpression of cyclooxygenase-2 (COX-2) and phosphorylation of nuclear factor-kappa B (NF-ĸB) p65 subunit. Inhibition of COX-2 by NS-398, a selective inhibitor of COX-2, significantly repressed the cytotoxicity, as well as secretion of IL-6 and IL-8 induced by CoCl2. Inhibition of NF-ĸB by PDTC (a selective inhibitor of NF-ĸB) or genetic silencing of p65 by RNAi (Si-p65), attenuated not only the cytotoxicity and secretion of IL-6 and IL-8, but also overexpression of COX-2 in CoCl2-treated HaCaT cells. Neutralizing anti-IL-6 or anti-IL-8 antibody statistically alleviated CoCl2-induced cytotoxicity in HaCaT cells. N-acetyl-L-cysteine (NAC), a well characterized ROS scavenger, obviously suppressed CoCl2-induced cytotoxicity in HaCaT cells, as well as secretion of IL-6 and IL-8. Additionally, NAC also repressed overexpression of COX-2 and phosphorylation of NF-ĸB p65 subunit induced by CoCl2 in HaCaT cells. In conclusion, our results demonstrated that oxidative stress mediates chemical hypoxia-induced injury and inflammatory response through activation of NF-ĸB?COX-2 pathway in HaCaT cells.

Keywords chemical hypoxia, cyclooxygenase-2, inflammation, keratinocytes, nuclear factor-kappa B, oxidative stress

Article

Research Article

Mol. Cells 2011; 31(6): 531-538

Published online June 30, 2011 https://doi.org/10.1007/s10059-011-1025-3

Copyright © The Korean Society for Molecular and Cellular Biology.

Oxidative Stress Mediates Chemical Hypoxia-Induced Injury and Inflammation by Activating NF-ĸb-COX-2 Pathway in HaCaT Cells

Chuntao Yang, Hongzhong Ling1, Meifen Zhang2, Zhanli Yang, Xiuyu Wang, Fanqin Zeng3, Chuhuai Wang4, and Jianqiang Feng*

Department of Physiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangdong, People’s Republic of China, 1Department of Dermatology, The First Affiliated Hospital, Sun Yat-sen University, Guangdong, People’s Republic of China, 2School of Nursing, Sun Yat-sen University, Guangdong, People’s Republic of China, 3Department of Dermatology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangdong, People’s Republic of China, 4Department of Rehabilitation, The First Affiliated Hospital, Sun Yat-sen University, Guangdong, People’s Republic of China

Correspondence to:*Correspondence: fengjq-sums@163.com

Received: February 14, 2011; Revised: March 21, 2011; Accepted: March 22, 2011

Abstract

Hypoxia of skin is an important physiopathological process in many diseases, such as pressure ulcer, diabet-ic ulcer, and varicose ulcer. Although cellular injury and inflammation have been involved in hypoxia-induced dermatic injury, the underlying mechanisms remain largely unknown. This study was conducted to investigate the effects of cobalt chloride (CoCl2), a hypoxia-mimicking agent, on human skin keratinocytes (HaCaT cells) and to explore the possible molecular mechanisms. Exposure of HaCaT cells to CoCl2 reduced cell viability and caused overproduction of reactive oxygen species (ROS) and oversecretion of interleukin-6 (IL-6) and interleukin-8 (IL-8). Importantly, CoCl2 exposure elicited overexpression of cyclooxygenase-2 (COX-2) and phosphorylation of nuclear factor-kappa B (NF-ĸB) p65 subunit. Inhibition of COX-2 by NS-398, a selective inhibitor of COX-2, significantly repressed the cytotoxicity, as well as secretion of IL-6 and IL-8 induced by CoCl2. Inhibition of NF-ĸB by PDTC (a selective inhibitor of NF-ĸB) or genetic silencing of p65 by RNAi (Si-p65), attenuated not only the cytotoxicity and secretion of IL-6 and IL-8, but also overexpression of COX-2 in CoCl2-treated HaCaT cells. Neutralizing anti-IL-6 or anti-IL-8 antibody statistically alleviated CoCl2-induced cytotoxicity in HaCaT cells. N-acetyl-L-cysteine (NAC), a well characterized ROS scavenger, obviously suppressed CoCl2-induced cytotoxicity in HaCaT cells, as well as secretion of IL-6 and IL-8. Additionally, NAC also repressed overexpression of COX-2 and phosphorylation of NF-ĸB p65 subunit induced by CoCl2 in HaCaT cells. In conclusion, our results demonstrated that oxidative stress mediates chemical hypoxia-induced injury and inflammatory response through activation of NF-ĸB?COX-2 pathway in HaCaT cells.

Keywords: chemical hypoxia, cyclooxygenase-2, inflammation, keratinocytes, nuclear factor-kappa B, oxidative stress

Mol. Cells
Feb 28, 2023 Vol.46 No.2, pp. 69~129
COVER PICTURE
The bulk tissue is a heterogeneous mixture of various cell types, which is depicted as a skein of intertwined threads with diverse colors each of which represents a unique cell type. Single-cell omics analysis untangles efficiently the skein according to the color by providing information of molecules at individual cells and interpretation of such information based on different cell types. The molecules that can be profiled at the individual cell by single-cell omics analysis includes DNA (bottom middle), RNA (bottom right), and protein (bottom left). This special issue reviews single-cell technologies and computational methods that have been developed for the single-cell omics analysis and how they have been applied to improve our understanding of the underlying mechanisms of biological and pathological phenomena at the single-cell level.

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