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 June 30, 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
Nov 30, 2022 Vol.45 No.11, pp. 763~867
COVER PICTURE
Naive (cyan) and axotomized (magenta) retinal ganglion cell axons in Xenopus tropicalis (Choi et al., pp. 846-854).

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