北京大学学报（医学版）2023，Vol.55Issue(6)：975-981,7.DOI:10.19723/j.issn.1671-167X.2023.06.004
干扰素-α介导系统性红斑狼疮外周血CD56dim CD57+自然杀伤细胞功能的损伤
Interferon-α mediating the functional damage of CD56dimCD57+natural killer cells in peripheral blood of systemic lupus erythematosuss
摘要
Abstract
Objective:To investigate the regulatory effect of interferon-α(IFN-α)on the apoptosis and killing function of CD56dimCD57+natural killer(NK)cells in systemic lupus erythematosus(SLE)patients,and to explore the specific mechanism.Methods:A total of sixty-four newly treated SLE pa-tients and sixteen healthy controls(HC)enrolled in the Second Hospital of Dalian Medical University were selected as the research subjects.And the gene expression levels of molecules related to NK cell-killing function were detected by real-time quantitative polymerase chain reaction.CD56dim CD57+NK cells were co-cultured with the K562 cells,and the apoptotic K562 cells were labeled with Annexin-Ⅴand 7-amino-actinomycin D.Peripheral blood mononuclear cells were treated with 20,40,and 80 μmol/L hydrogen peroxide(H2 O2),and treated without H2 O2 as control,the expression level of perforin(PRF)was detected by flow cytometry.The concentration of IFN-α in serum was determined by enzyme linked immunosorbent assay.The expression levels of IFN-α receptors(IFNAR)on the surface of CD56dimCD57+NK cells were detected by flow cytometry,and were represented by mean fluorescence in-tensity(MFI).CD56dimCD57+NK cells were treated with 1 000 U/mL IFN-α for 24,48 and 72 h,and no IFN-α treatment was used as the control,the apoptosis and the expression levels of mitochondrial reac-tive oxygen species(mtROS)were measured by flow cytometry and represented by MFI.Results:Com-pared with HC(n=3),the expression levels of PRF1 gene in peripheral blood NK cells of the SLE pa-tients(n=3)were decreased(1.24±0.41 vs.0.57±0.12,P=0.05).Compared with HC(n=5),the ability of peripheral blood CD56dimCD57+NK cells in the SLE patients(n=5)to kill K562 cells was significantly decreased(58.61％±10.60％vs.36.74％±6.27％,P＜0.01).Compared with the control(n=5,97.51％±1.67％),different concentrations of H2O2 treatment significantly down-regu-lated the PRF expression levels of CD56dimCD57+NK cells in a dose-dependent manner,the 20 μmol/L H2O2 PRF was 83.23％±8.48％(n=5,P＜0.05),the 40 μmol/L H2O2 PRF was 79.53％±8.56％(n=5,P＜0.01),the 80 μmol/L H2O2 PRF was 76.67％±7.16％(n=5,P＜0.01).Compared to HC(n=16),the serum IFN-α levels were significantly increased in the SLE patients(n=45)with moderate to high systemic lupus erythematosus disease activity index(SLEDAI≥10)[(55.07±50.36)ng/L vs.(328.2±276.3)ng/L,P＜0.001].Meanwhile,compared with HC(n=6),IFNAR1 ex-pression in peripheral blood CD56dimCD57+NK cells of the SLE patients(n=6)were increased(MFI:292.7±91.9 vs.483.2±160.3,P＜0.05),and compared with HC(n=6),IFNAR2 expression in peripheral blood CD56dimCD57+NK cells of the SLE patients(n=7)were increased(MFI:643.5± 113.7 vs.919.0±246.9,P＜0.05).Compared with control(n=6),the stimulation of IFN-α(n=6)significantly promoted the apoptosis of CD56dimCD57+NK cells(20.48％±7.01％vs.37.82％± 5.84％,P＜0.05).In addition,compared with the control(n=4,MFI:1 049±174.5),stimulation of CD56dimCD57+NK cells with IFN-α at different times significantly promoted the production of mtROS in a time-dependent manner,48 h MFI was 3 437±1 472(n=4,P＜0.05),72 h MFI was 6 495± 1 089(n=4,P＜0.000 1),but there was no significant difference at 24 h of stimulation.Conclusion:High serum IFN-α level in SLE patients may induce apoptosis by promoting mtROS production and inhibit perforin expression,which can down-regulate CD56 dimCD57+NK killing function.关键词
系统性红斑狼疮/自然杀伤细胞/干扰素-α/活性氧/穿孔素Key words
Systemic lupus erythematosus/Natural killer cells/Interferon-α/Reactive oxygen spe-cies/Perforin分类
医药卫生引用本文复制引用
赵祥格,刘佳庆,黄会娜,陆智敏,白自然,李霞,祁荆荆..干扰素-α介导系统性红斑狼疮外周血CD56dim CD57+自然杀伤细胞功能的损伤[J].北京大学学报（医学版）,2023,55(6):975-981,7.基金项目
国家自然科学基金(82201990、82071834、82271839)和大连市青年科技之星项目(2022RQ070)Supported by the National Natural Science Foundation of China(82201990,82071834,82271839)and Science and Technology Talent Innovation Support Project of Dalian(2022RQ070) （82201990、82071834、82271839）
