青蒿素调节Tregs中NRP1/PTEN/Foxp3信号通路减轻系统性红斑狼疮小鼠炎性反应的研究  

Artesunate attenuates the inflammatory response in systemic lupus erythematosus mice by regulation loop of the NRP1/PTEN/Foxp3 signaling pathway in regulatory T cells

作　　者：洪学志 朱浈臻 王亚慧 莫海露 刘雷 莫汉有[1] HONG Xuezhi;ZHU Zhenzhen;WANG Yahui;MO Hailu;LIU Lei;MO Hanyou(Department of Clinical Immunology and Rheumatology,The Affiliated Hospital of Guilin Medical College,Guilin,Guangxi 541004,China)

机构地区：[1]桂林医学院附属医院风湿免疫科,广西桂林541004

出　　处：《重庆医学》2023年第12期1776-1782,共7页Chongqing medicine

基　　金：国家自然科学基金项目(81760298);广西壮族自治区自然科学基金项目(2020GXNSFBA297149);广西壮族自治区研究生教育创新计划项目(YCSW2022375)。

摘　　要：目的分析青蒿素(ART)对调节性T淋巴细胞(Tregs)功能的影响及机制,并评估其在系统性红斑狼疮(SLE)小鼠模型中的治疗潜力。方法(1)使用生物信息学方法分析叉状头/翅膀状螺旋转录因子3(Foxp3)基因缺失的Tregs与Foxp3+Tregs中差异性表达基因(DEGs)并行DEGs富集通路分析。利用String数据库进一步明确神经纤毛蛋白-1(NRP1)、人第10号染色体缺失的磷酸酶及张力蛋白同源蛋白(PTEN)、Foxp3之间的关系。同时使用ZDOCK进行PTEN与NRP1物理对接。(2)体内实验中将降植烷诱导的SLE小鼠分别使用0.4%甲基纤维素(对照组)、PTEN抑制剂(SF1670)和ART腹腔注射28 d。比较各组外周血清双链DNA(dsDNA)、抗核抗体(ANA)、促炎因子[白细胞介素(IL)-6、IL-1、IL-23、IL-17]及抗炎因子(IL-10)的表达水平及脾脏淋巴细胞中Tregs占比。另将纯化的外周血Tregs予0.1%二甲基亚砜、SF1670和ART分别干预24 h,比较不同组细胞中磷酸化PTEN(pPTEN)表达水平。结果通过生物信息学分析发现,Foxp3基因表达与NRP1和PTEN有关,且这3个蛋白之间存在相互作用,且NRP1与PTEN具有物理接触。体外实验结果显示:与对照组相比,ART组的NRP1(P>0.05)和pPTEN(P<0.05)表达上调;ART明显降低了SLE小鼠体内促炎症因子、ANA和dsDNA表达水平,并明显提高了Tregs占比,降低了效应性T淋巴细胞占比,而使用PTEN抑制剂(SF1670)则出现与ART相反的结果。结论ART对Tregs的诱导作用与NRP1、PTEN活性升高有关,ART可能通过调节NRP1/PTEN/Foxp3信号通路减轻SLE小鼠的炎性反应。Objective To investigate the effects and mechanisms of artemisinin(ART)on regulatory T lymphocyte(Tregs)function and its therapeutic potential in a mouse model of systemic lupus erythematosus(SLE).Methods(1)Differential expression genes(DEGs)analysis and pathway enrichment analysis were conducted on Tregs with forkhead/winged helix familytranscriptionfactor 3(Foxp3)gene deletion compared to Foxp3+Tregs using bioinformatics.The relationship between NRP1,PTEN,and Foxp3 proteins was further examined using the String database,and physical docking of PTEN protein to NRP1 protein was performed by using ZDOCK.(2)In vivo experiments,the SLE mice induced by hypoglycan were intraperitoneally injected with 0.4%methylcellulose(the control group),PTEN inhibitor(SF1670),and ART for 28 days,respectively.The levels of peripheral serum double-stranded DNA(dsDNA),antinuclear antibody(ANA),pro-inflammatory factors,including interleukin(IL)-6,IL-1,IL-23 and IL-17,anti-inflammatory factor(IL-10),and the percentage of Tregs in spleen lymphocytes were compared between the different treatment groups.Furthermore,the expression levels of phosphorylated PTEN(pPTEN)in peripheral blood Tregs were compared among purified cells treated with 0.1%dimethyl sulfoxide,SF1670,and ART for 24 hours in an in vitro assay.Results The bioinformatics analysis revealed an association between Foxp3 gene expression and NRP1 and PTEN genes,and a physical interaction between NRP1 and PTEN proteins was observed.In the in vitro experiments,the expression of NRP1(P>0.05)and pPTEN(P<0.05)was increased in the ART group compared to the control group.The in vivo experiments demonstrated that ART significantly decreased the expression levels of pro-inflammatory factors,ANA,and dsDNA in SLE mice.Additionally,ART significantly increased the percentage of Tregs and decreased the percentage of effector T cells.Conversely,the use of the PTEN inhibitor(SF1670)yielded opposite results compared to ART.Conclusion The induction of Tregs by ART was associated with increased NRP1

关 键 词：系统性红斑狼疮 调节性T细胞 青蒿素 NRP1 PTEN 

分 类 号：R285.5[医药卫生—中药学]