氧化应激促进MHCⅠ类分子膜转位在CD8+T细胞杀伤黑素细胞中的作用  

Membrane Translocation of MHC Class Ⅰ Promotes Melanocyte Damage by CD8^(+) T Cells Under Oxidative Stress

作　　者：毛汉潇 王婧莹 曾紫原 潘映浩 刘汝兰 熊霞 何渊民 MAO Hanxiao;WANG Jingying;ZENG Ziyuan;PAN Yinghao;LIU Rulan;XIONG Xia;HE Yuanmin(Department of Dermatology,Affiliated Hospital of Southwest Medical University,Luzhou 646000 China)

机构地区：[1]西南医科大学附属医院皮肤科,四川泸州646000

出　　处：《中国皮肤性病学杂志》2023年第11期1232-1236,1245,共6页The Chinese Journal of Dermatovenereology

基　　金：四川省科技创新创业苗子工程项目(2022JDRC0147);泸州市人民政府-西南医科大学科技战略合作项目(2019LZXNYDJ03)。

摘　　要：目的 探索氧化应激诱导下主要组织相容性复合体(MHC)Ⅰ类分子膜转位在CD8^(+)T细胞靶向杀伤黑素细胞中的作用。方法 用1.0 mmol/L的H_(2)O_(2)处理正常人原代黑素细胞,采用流式细胞术检测黑素细胞内活性氧(ROS)水平、黑素细胞和CD8^(+)T细胞共培养体系中颗粒酶、穿孔素表达水平,Western blot检测黑素细胞膜表面MHCⅠ类分子表达情况。用MHC class I siRNA干涉黑素细胞MHCⅠ类分子表达后,用流式细胞术检测黑素细胞和CD8+T细胞共培养体系中颗粒酶、穿孔素表达水平。免疫荧光检测黑素细胞膜、正常人皮肤及白癜风皮损周围MHCⅠ类分子表达情况。结果 经1.0 mmol/L H2O2处理黑素细胞24 h后,黑素细胞膜表面MHCⅠ类分子表达显著升高;同时,黑素细胞和CD8^(+)T细胞共培养体系中黑素细胞坏死比例增高,颗粒酶B、穿孔素和TNF-α水平明显升高;经1.0 mmol/L H_(2)O_(2)处理干涉MHCⅠ类分子后的黑素细胞24 h,与CD8^(+)T细胞共培养,共培养体系中颗粒酶B、穿孔素和TNF-α释放水平明显降低,黑素细胞坏死比例下降。结论 氧化应激可通过促进MHCⅠ类分子向胞膜转位,诱导CD8^(+)T细胞杀伤黑素细胞。Objective To explore the effect of MHC classⅠmolecules membrane translocation in the target killing of melanocytes by CD8^(+)T cells under oxidative stress.MethodsOxidative stress was induced in normal human primary epidermal melanocytes by treatment with 1.0 mmol/L H_(2)O_(2).ROS levels in melanocytes,granzyme and perforin in melanocyte and CD8^(+)T cellco-culture system were detected by flow cytometry.The expression of MHC classⅠmolecules on melanocyte membrane was detected by Western blot.After interfering with the expression of MHC class I in melanocytes with MHC classⅠsiRNA,the expression of granzyme and perforin in melanocytes and CD8^(+)T cellco-culture system was detected by flow cytometry.The expression of MHC classⅠmolecules in melanocyte membrane,normal human skin and vitiligo lesions was detected by immunofluorescence.Results The expression of MHC classⅠmolecules on the cell membrane increased after melanocytes were treated with 1.0 mmol/L H_(2)O_(2) for 24 h.The proportions of melanocyte necrosis and the levels of granzyme B,perforin and TNF-αwere significantly increased in the co-culture system of melanocytes and CD8^(+)T cells.After MHC classⅠmolecules knock-down in melanocytes,the levels of granzyme B,perforin and TNF-αwere significantly reduced in the co-culture system,and the proportion of melanocyte necrosis decreased.Conclusion Oxidative stress can induce the transposition of MHC classⅠmolecules to cell membrane and promote CD8^(+)T cells to kill melanocytes.

关 键 词：氧化应激 黑素细胞 MHCⅠ类分子 

分 类 号：R751[医药卫生—皮肤病学与性病学]