维生素D3通过激活Nrf2抑制氮芥诱导角质形成细胞铁死亡的作用研究  

Vitamin D3 inhibits nitrogen mustard-induced keratinocyte ferroptosis by activating Nrf2

作　　者：董训虎 刘浩银 葛维 陈明亮[1,2] DONG Xunhu;LIU Haoyin;GE Wei;CHEN Mingliang(Department of Chemical Defense,Faculty of Military Preventive Medicine,Army Medical University(Third Military Medical University),Chongqing,China;State Key Laboratory of Trauma and Chemical Poisoning,Faculty of Military Preventive Medicine,Army Medical University(Third Military Medical University),Chongqing,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,重庆 [2]陆军军医大学(第三军医大学)创伤与化学中毒全国重点实验室,重庆

出　　处：《陆军军医大学学报》2025年第7期674-680,共7页Journal of Army Medical University

基　　金：创伤与化学中毒全国重点实验室开放基金项目(SKL0202407)。

摘　　要：目的探索铁死亡在维生素D3(vitamin D3,VD3)改善氮芥(nitrogen mustard,NM)诱导角质形成细胞(HaCaT)毒性损伤中的作用及机制。方法采用梯度浓度NM(5、10、20和50μmol/L)单独或联合铁死亡抑制剂ferrostatin-1或liproxstatin-1(10μmol/L)、核因子-E2相关因子2(nuclear factor E2 related factor 2,Nrf2)特异性激动剂tBHQ(10μmol/L)、VD3(10 nmol/L)处理HaCaT细胞。共培养24 h后,分别检测细胞增殖活力、细胞内谷胱甘肽(glutathione,GSH)和脂质过氧化物(malondialdehyde,MDA)含量,以及Nrf2、xCT和GPX4等蛋白的表达水平。结果NM以浓度依赖性方式抑制HaCaT细胞增殖活力,当NM浓度为20μmol/L时,细胞增殖活力下降至约55%(P<0.05)。铁死亡抑制剂能够减轻NM诱导的HaCaT细胞毒性损伤(P<0.05)。此外,激活Nrf2可显著抑制NM对HaCaT细胞铁死亡的诱导效应,表现为细胞内GSH水平的恢复和MDA含量的降低(P<0.05)。同时,VD3能够靶向激活Nrf2信号通路,上调xCT和GPX4蛋白的表达,从而有效抑制HaCaT细胞的铁死亡,并缓解NM诱导的细胞毒性。结论VD3通过激活Nrf2抑制HaCaT细胞铁死亡,进而改善NM诱导的细胞毒性损伤。Objective To explore the role and mechanism of ferroptosis in vitamin D3(VD3)improving the cytotoxicity of keratinocytes(HaCaT cells)induced by nitrogen mustard(NM).Methods HaCaT cells were treated with gradient concentrations of NM(5,10,20,and 50μmol/L)alone or in combination with ferroptosis inhibitor ferrostatin-1 or liproxstatin-1(10μmol/L),Nrf2-specific agonist tBHQ(10μmol/L),and VD3(10 nmol/L),respectively.After co-culture for 24 h,cell viability was assessed,and the intracellular contents of glutathione(GSH)and malondialdehyde(MDA)were measured.Additionally,the expression levels of Nrf2,xCT,and GPX4 were detected.Results NM exerted inhibitory effect on the proliferation of HaCaT cells in a dose-dependent manner,and the cell viability was reduced to approximately 55%at an NM concentration of 20μmol/L(P<0.05).Ferroptosis inhibitors significantly mitigated the cytotoxic damage induced by NM in HaCaT cells(P<0.05),but activation of nuclear factor E2 related factor 2(Nrf2)markedly attenuated NM-induced ferroptosis,as indicated by the restoration of intracellular GSH level and the decrease in MDA content(P<0.05).VD3 specifically targeted the Nrf2 signaling pathway,upregulated the expression of xCT and GPX4 protein,thereby inhibiting ferroptosis and reducing NM-induced cytotoxicity in HaCaT cells.Conclusion VD3 mitigates NM-induced cytotoxicity in HaCaT cells by inhibiting ferroptosis via Nrf2 activation.

关 键 词：氮芥 维生素D3 核因子-E2相关因子2 角质形成细胞 铁死亡 

分 类 号：R963[医药卫生—微生物与生化药学] R977.24[医药卫生—药理学] R9964[医药卫生—药学]