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 (H₂S) has been shown to protect against oxidative stress injury and inflammation in various hypoxia-induced insult models. However, it remains unknown whether H₂S protects human skin keratinocytes (HaCaT cells) against chemical hypoxia-induced damage. In the current study, HaCaT cells were treated with cobalt chloride (CoCl₂), 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 H₂S) for 30 min before exposure to CoCl₂ for 24 h significantly attenuated CoCl₂-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 CoCl₂-induced COX-2 overexpression and PGE₂ secretion as well as intranuclear NF-κB p65 subunit accumulation (the central step of NF-κB activation). Similar to the protective effect of H₂S, both NS-398 (a selective COX-2 inhibitor) and PDTC (a selective NF-κB inhibitor) depressed not only CoCl₂-induced cytotoxicity, but also the secretions of IL-1β, IL-6 and IL-8. Importantly, PDTC obviously attenuated overexpression of COX-2 induced by CoCl₂. Notably, NAC, a ROS scavenger, conferred a similar protective effect of H₂S against CoCl₂-induced insults and inflammatory responses. Taken together, the findings of the present study have demonstrated for the first time that H₂S protects HaCaT cells against CoCl₂-induced injuries and inflammatory responses through inhibition of ROS-activated NF-κB/COX-2 pathway.