title: Hydrogen sulfide protects against chemical hypoxia-induced cytotoxicity and inflammation in HaCaT cells through inhibition of ROS/NF-κB/COX-2 pathway.
abstract: Hydrogen sulfide (H(2)S) has been shown to protect against oxidative stress injury and inflammation in various hypoxia-induced insult models. However, it remains unknown whether H(2)S protects human skin keratinocytes (HaCaT cells) against chemical hypoxia-induced damage. In the current study, HaCaT cells were treated with cobalt chloride (CoCl(2)), a well known hypoxia mimetic agent, to establish a chemical hypoxia-induced cell injury model. Our findings showed that pretreatment of HaCaT cells with NaHS (a donor of H(2)S) for 30 min before exposure to CoCl(2) for 24 h significantly attenuated CoCl(2)-induced injuries and inflammatory responses, evidenced by increases in cell viability and GSH level and decreases in ROS generation and secretions of IL-1β, IL-6 and IL-8. In addition, pretreatment with NaHS markedly reduced CoCl(2)-induced COX-2 overexpression and PGE(2) secretion as well as intranuclear NF-κB p65 subunit accumulation (the central step of NF-κB activation). Similar to the protective effect of H(2)S, both NS-398 (a selective COX-2 inhibitor) and PDTC (a selective NF-κB inhibitor) depressed not only CoCl(2)-induced cytotoxicity, but also the secretions of IL-1β, IL-6 and IL-8. Importantly, PDTC obviously attenuated overexpression of COX-2 induced by CoCl(2). Notably, NAC, a ROS scavenger, conferred a similar protective effect of H(2)S against CoCl(2)-induced insults and inflammatory responses. Taken together, the findings of the present study have demonstrated for the first time that H(2)S protects HaCaT cells against CoCl(2)-induced injuries and inflammatory responses through inhibition of ROS-activated NF-κB/COX-2 pathway.
authors.affiliation: Department of Physiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China.
authors.forename: Chuntao
authors.initials: C
authors.lastname: Yang
authors.forename: Zhanli
authors.initials: Z
authors.lastname: Yang
authors.forename: Meifen
authors.initials: M
authors.lastname: Zhang
authors.forename: Qi
authors.initials: Q
authors.lastname: Dong
authors.forename: Xiuyu
authors.initials: X
authors.lastname: Wang
authors.forename: Aiping
authors.initials: A
authors.lastname: Lan
authors.forename: Fanqin
authors.initials: F
authors.lastname: Zeng
authors.forename: Peixi
authors.initials: P
authors.lastname: Chen
authors.forename: Chuhuai
authors.initials: C
authors.lastname: Wang
authors.forename: Jianqiang
authors.initials: J
authors.lastname: Feng
mesh_terms: D015153:Blotting, Western; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003035:Cobalt / Q000494:pharmacology; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006862:Hydrogen Sulfide / Q000494:pharmacology*; D000860:Hypoxia / Q000139:chemically induced*; D053583:Interleukin-1beta / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009578:Nitrobenzenes / Q000494:pharmacology; D011392:Proline / Q000031:analogs & derivatives / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D013449:Sulfonamides / Q000494:pharmacology; D013859:Thiocarbamates / Q000494:pharmacology
chemical_list: D016861:Cyclooxygenase Inhibitors; D053583:Interleukin-1beta; D015850:Interleukin-6; D016209:Interleukin-8; D016328:NF-kappa B; D009578:Nitrobenzenes; D017382:Reactive Oxygen Species; D013449:Sulfonamides; D013859:Thiocarbamates; C080955:N-(2-cyclohexyloxy-4-nitrophenyl)methanesulfonamide; C066229:prolinedithiocarbamate; D003035:Cobalt; D011392:Proline; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; C018021:cobaltous chloride; D006862:Hydrogen Sulfide