Hydrogen sulfide protects against chemical hypoxia-induced cytotoxicity and inflammation in HaCaT cells through inhibition of ROS/NF-κB/COX-2 pathway

• Published in: PloS one 2011, Vol.6 (7), p.e21971
• Main Authors: Yang, Chuntao, Yang, Zhanli, Zhang, Meifen, Dong, Qi, Wang, Xiuyu, Lan, Aiping, Zeng, Fanqin, Chen, Peixi, Wang, Chuhuai, Feng, Jianqiang
• Format: Article
• Language: English
• Subjects: Biology; Blotting, Western; Cardiomyocytes; Cell injury; Cell Line, Tumor; Cell Survival - drug effects; Chemical damage; Cobalt; Cobalt - pharmacology; Cobalt chloride; COX-2 inhibitors; Cyclooxygenase 2 - metabolism; Cyclooxygenase Inhibitors - pharmacology; Cyclooxygenase-2; Cytokines; Cytotoxicity; Dermatology; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Gangrene; Homeostasis; Hospitals; Humans; Hydrogen; Hydrogen sulfide; Hydrogen Sulfide - pharmacology; Hypoxia; Hypoxia - chemically induced; Immune system; Inflammation; Inhibition; Injuries; Interleukin 6; Interleukin 8; Interleukin-1beta - metabolism; Interleukin-6 - metabolism; Interleukin-8 - metabolism; Ischemia; Keratinocytes; Kinases; Medicine; NF-kappa B - antagonists & inhibitors; NF-kappa B - metabolism; NF-κB protein; Nitrobenzenes - pharmacology; Oxidative stress; Physiology; Pressure ulcers; Proline - analogs & derivatives; Proline - pharmacology; Prostaglandin E2; Proteins; Reactive Oxygen Species - metabolism; Rodents; Secretions; Signal Transduction - drug effects; Skin; Sulfide; Sulfonamides - pharmacology; Thiocarbamates - pharmacology; Toxicity
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0021971

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.