维生素D3通过抑制NLRP3炎性小体激活改善氮芥诱导角质形成细胞炎性损伤  

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

作　　者：董训虎 陈明亮[1] 余文珮 叶枫[1] 但国蓉[1] 邹仲敏[1] DONG Xunhu;CHEN Mingliang;YU Wenpei;YE Feng;DAN Guorong;ZOU Zhongmin(Department of Chemical Defense,Faculty of Military Preventive Medicine,Army Medical University(Third Military Medical University),Chongqing,400038,China)

机构地区：[1]陆军军医大学(第三军医大学)军事预防医学系防化医学教研室

出　　处：《第三军医大学学报》2020年第3期267-272,共6页Journal of Third Military Medical University

基　　金：国家自然科学基金青年科学基金(81703268,81803279)~~

摘　　要：目的探讨NLRP3炎性小体在维生素D3(vitamin D3,VD3)改善氮芥(nitrogen mustard, NM)诱导角质形成细胞炎性损伤中的作用和机制。方法用20μmol/L NM单独或联合用10μmol/L caspase-1特异抑制剂(Ac-YVAD-cmk, YVAD)、10μmol/L NLRP3炎性小体特异抑制剂(MCC950)、5 nmol/L VD3处理人角质形成细胞株(HaCaT细胞)4 h。应用CCK-8法检测细胞增殖活力,Western blot检测NLRP3、caspase-1 p20、IL-1β和COX-2的表达水平,ELISA检测IL-1β释放量。结果 20μmol/L NM对HaCaT细胞增殖活力无显著影响,NM处理后可显著上调HaCaT细胞NLRP3、caspase-1 p20、IL-1β和COX-2的蛋白表达水平,增加细胞IL-1β释放(P<0.05)。而加入MCC950或YVAD处理后能显著抑制NM诱导的上述效应(P<0.05);同时,VD3干预能够明显抑制NM对NLRP3和caspase-1 p20蛋白表达的上调作用,并显著减少NM诱导的IL-1β和COX-2的表达和释放(P<0.05)。结论 VD3通过抑制NLRP3炎性小体激活减少角质形成细胞炎性因子生成,改善NM诱导的细胞炎性损伤。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.

关 键 词：氮芥 维生素D3 NLRP炎性小体 角质形成细胞 炎性反应 

分 类 号：R827.174[医药卫生—临床医学] R967