Vitamin D3 ameliorates nitrogen mustard-induced keratinocytes inflammation by inactivating NLRP3 inflammasome activation

Abstract

Objective To investigate the effect of vitamin D3 on nitrogen mustard (NM)-induced keratinocyte inflammation and the role of NOD-like receptor protein 3 (NLRP3) inflammasome in the process. Methods HaCaT cells were treated with 20 μmol/L NM, in the presence or absence of 10 μmol/L MCC950 (a specific inhibitor of NLRP3 inflammasome) or 10 μmol/L Ac-YVAD-cmk (YVAD, a specific activator of caspase-1) or 5 nmol/L vitamin D3 for 4 h. CCK-8 assay was used to detect cell viability. The IL-1β content in the supernatant of cell lysate was determined by Human IL-1β ELISA kit, and the expression of NLRP3, caspase-1, IL-1β and COX-2 was detected by Western blotting. Results Treatment with 20 μmol/L NM had no significant effect on the viability of HaCaT cells, but remarkably up-regulated the expression of NLRP3, caspase-1 p20, IL-1β and COX-2, and enhanced the accumulation of IL-1β in the culture supernatant of HaCaT cells (P < 0.05). Moreover, MCC950 or YVAD treatment abolished the above effects of NM (P < 0.05). Simultaneously, vitamin D3 treatment notably inhibited the upregulated expression of NLRP3 and caspase-1 p20, and remarkably reduced the increased expression and release of IL-1β and COX-2 in HaCaT cells induced by NM (P < 0.05). Conclusion Vitamin D3 ameliorates NM-induced keratinocytes inflammation by reducing the synthesis of inflammatory mediators through suppressing the activation of NLRP3 inflammasome

Keywords

• nitrogen mustard
• vitamin d3
• nod-like receptor protein 3 inflammasome
• keratinocytes
• inflammatory response