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
Chuntao Yang, Hongzhong Ling1, Meifen Zhang2, Zhanli Yang, Xiuyu Wang, Fanqin Zeng3, Chuhuai Wang4, and Jianqiang Feng*
Correspondence to : *Correspondence: fengjq-sums@163.com
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 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.
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
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
Samar Basu*
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