二甲双胍调节系统性效应T细胞/调节性T细胞平衡对博来霉素诱导系统性硬化病小鼠模型的影响  

Effect of metformin on bleomycin-induced systemic sclerosis mice by regulating the balance of Teff/regulatory T cells

作　　者：梁朝 王艳林 赵向聪[1] 范春雪 罗静[1] Liang Zhaojun;Wang Yanlin;Zhao Xiangcong;Fan Chunxue;Luo Jing(Department of Rheumatology,The Second Hospital of Shanxi Medical University,Taiyuan 030001,China)

机构地区：[1]山西医科大学第二医院风湿免疫科,太原030001

出　　处：《中华风湿病学杂志》2021年第7期461-466,I0003,共7页Chinese Journal of Rheumatology

基　　金：山西省重点研发计划(国际合作领域)项目(201803D421067);山西省应用基础研究计划项目(201901D111377);山西省卫生健康委科研课题(2019044);山西省回国留学人员科研教研项目(2020-191)。

摘　　要：目的研究不同浓度二甲双胍对博来霉素(BLM)诱导SSc小鼠模型的影响及机制。方法C57BL/6小鼠按照随机分配原则分为正常组、模型组、高、中、低剂量二甲双胍治疗组,经博来霉素、二甲双胍干预4周末处死小鼠。局部皮肤采用组织病理染色法测量真皮和胶原厚度,免疫组织化学法和实时荧光定量PCR(RT-qPCR)检测IL-17、叉头框蛋白P3(Foxp3)、α平滑肌肌动蛋白(α-SMA)的表达情况和mRNA水平。采用流式细胞术检测脾单个核细胞中系统性效应T细胞(Teff)和调节性T细胞(Treg)相对数目。对真皮胶原厚度、α-SMA、IL-17、Foxp3、Teff和Treg水平等数据均采用单因素方差分析进行统计分析,两两比较采用LSD-t检验或Kruskal-wallis检验。结果与正常组相比,模型组小鼠出现明显的皮肤病变,Th1、Th2、Th17、Tfh细胞水平明显升高,同时伴有Treg水平明显下降。经高剂量二甲双胍治疗后,小鼠真皮厚度[(131±25)μm]、胶原厚度[(119±18)μm]、α-SMA[(3.0±0.5)个/高倍视野]明显减少(F=14.390,P<0.01;F=40.245,P<0.01;F=44.626,P<0.01),Th1[(27.00±6.68)%]、Th17[(0.56±0.20)%]、Tfh[(6.4±1.6)%]细胞水平明显降低(F=32.390,P<0.01;F=16.083,P<0.01;F=16.546,P<0.01),Treg[(11.23±1.52)%]水平明显升高(F=10.171,P<0.01)。结论二甲双胍可以有效逆转博来霉素诱导SSc小鼠模型的皮肤局部症状,并通过恢复Teff/Treg平衡达到免疫调节和抗纤维化作用。Objective To explore the effect and mechanism of different concentrations of metformin on bleomycin(BLM)-induced systemic sclerosis(SSc)mice model.Methods C57BL/6 mice were divided into the normal group,the model group,the high,the medium and the low metformin(MET)treatment groups randomly.All mice were sacrificed after BLM and metformin treatment for 4 weeks.Local skin was exminedby histopathological staining method to measure the thickness of dermis and collagen,and immunohistochemistry and real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)were used to detect the protein and mRNA levels of Interleukin(IL)-17,forkhead box P3(Foxp3)andα-smooth muscle actin(α-SMA).Flow cytometry was used to detect the percentage of effector T cell(Teff)and regulatory cells(Treg)in splenic mononuclear cells.The data such as dermal collagen thickness,α-SMA,IL-17,Foxp3,Teff and Treg levels were statistically analyzed by one-way analysis of variance.The data such as dermal collagen thickness,α-SMA,IL-17,Foxp3,Teff and Treg levels were analyzed by one-way analysis of variance,and least significant difference(LSD)-t or Kruskal-Wallis test was used for comparison between groups.Results Compared with the normal group,remarkable fibrotic lesions appeared in the skin of mice in the model group,and the levels of T-helper cells(Th)1,Th2,Th17,and T follicular helper cells(Tfh)cells were increased,accompanied by a significant decrease in the level of Treg cells.After high-dose metformin treatment,the dermal thickness[(131±25)μm],collagen thickness[(119±18)μm],andα-SMA[(3.0±0.5)/HPH]were significantly reduced(F=14.390,P<0.01;F=40.245,P<0.01;F=44.626,P<0.01).Th1[(27.00±6.68)%],Th17[(0.56±0.20)%],Tfh[(6.4±1.6)%]cells ware significantlyreduced(F=32.390,P<0.01;F=16.083,P<0.01;F=16.546,P<0.01),and Treg[(11.23±1.52)%]cells were significantly increased(F=10.171,P<0.01).Conclusion Metformin can effectively reverse the local skin changesin BLM-induced SSc mouse model,and show immune regulation and anti-fibrosis effects b

关 键 词：二甲双胍 T淋巴细胞 调节性 免疫调节 硬化病 系统性 效应T细胞 

分 类 号：R965[医药卫生—药理学]