辅助性T细胞17/调节性T细胞失衡在大鼠自身免疫性肝炎模型中的研究  

Study on the imbalance between T helper cell 17 and regulatory cells in autoimmune hepatitis model of rats

作　　者：高丽萍 毛小荣 GAO Li-ping;MAO Xiao-rong(Department of Infectious Diseases,The First Hospital of Lanzhou University Medical College,Lanzhou 730000,Gansu Province,China)

机构地区：[1]兰州大学医学院第一医院感染科,甘肃兰州730000

出　　处：《中国临床药理学杂志》2020年第19期3021-3025,共5页The Chinese Journal of Clinical Pharmacology

基　　金：甘肃省自然科学基金资助项目(1606RJZA127)。

摘　　要：目的研究辅助性T细胞17(Th17)和调节性T细胞(Treg)失衡在Wistar大鼠急、慢性免疫性肝炎模型中的意义和作用。方法将30只雌性Wistar大鼠随机分为对照组(0.9%NaCl),实验组-1[α-半乳糖酰基鞘氨醇(α-galcer)40μg·kg^-1],实验组-2[刀豆蛋白A(ConA)12.5 mg·kg^-1]。给药7周后,检测肝功能生化指标;用苏木精-伊红(HE)染色法观察病理改变情况;用免疫组织化学方法检测叉头盒蛋白3(Foxp3)、维A酸相关孤儿核受体γt(RORγt)蛋白表达情况;用酶联免疫吸附实验法检测外周血白细胞介素(IL)-17 A、IL-6、IL^-10、转化生长因子-β1(TGF-β1)细胞因子的含量;用流式细胞分析术检测Th17和Treg频率变化,并计算Th17/Treg比值。结果对照组、实验组-1、实验组-2血清中谷丙转氨酶(GPT)分别为(48.44±27.00),(82.50±8.61)和(278.96±34.44)U·L^-1;谷草转氨酶(GOT)分别为(95.02±22.10),(186.08±72.36)和(293.64±27.26)U·L^-1;血清总胆红素(TBiL)分别为(0.86±0.42),(1.82±0.78)和(14.88±7.67)μmol·L^-1;白蛋白(ALB)分别为(34.64±1.90),(27.13±4.26)和(21.55±2.65)g·L^-1;IL^-17A分别为(39.84±21.04),(68.24±13.01)和(238.11±32.80)pg·mL^-1;IL-6分别为(69.23±14.24),(91.84±5.62)和(99.09±22.44)pg·mL^-1;IL^-10分别为(53.83±9.14),(81.67±15.26)和(99.38±25.98)pg·mL^-1;TGF-β1分别为(105.86±24.98),(161.86±29.89)和(219.73±59.68)pg·mL^-1;对照组、实验组-1、实验组-2外周血中Th17细胞频率分别为(1.33±0.20)%,(5.55±1.57)%和(3.06±0.56)%;Treg细胞频率分别为(2.53±0.42)%,(4.96±0.61)%和(12.05±1.00)%;Th17/Treg比率分别为0.52±0.31,1.12±0.30和0.25±0.04;对照组、实验组-1、实验组-2肝组织中Foxp3蛋白表达(IOD值)分别为(1.67±0.12),(1.36±0.11)和(1.99±0.14);对照组分别与实验组-1与实验组-2相比,差异均有统计学意义(均P<0.05)。结论Th17/Treg比值在急性免疫性肝炎模型中明显上升,在慢性自身免疫性肝炎模型中明显下降,Th17/Treg的失衡参与自身免疫性肝炎�Objective To study the significance and role of imbalance between helper T cells 17(Th17)and regulatory T cells(Treg)in acute and chronic immune hepatitis in rats.Methods Thirty female Wistar rats were randomly divided into control group(0.9%NaCl),test-1 group[40μg·kg^-1 of(α-galactosylsphingosine(α-GalCer)]and test-2 group[12.5 mg·kg^-1 of concanavalin A(ConA)].After 7 weeks of administration,the biochemical indexes of liver function were detected;the pathological changes were observed by hematoxylin-eosin staining;the expression of forkhead box p3(Foxp3)and retinoid-related orphan nuclear receptorγt(RORγt)protein were detected by immunohistochemistry;the contents of interleukin(IL)-17 A,IL-6,IL^-10 and transforming growth factor-β1(TGF-β1)cytokines in peripheral blood were detected by enzyme linked immunosorbent assay;the frequency changes of Th17 and Tregs were detected by flow cytometry,and the ratio of Th17/Treg was calculated.Results The levels of glutamyl oxaloacetic transaminase(GPT)in serum of control group,test-1 and test-2 groups were(48.44±27.00),(82.50±8.61)and(278.96±34.44)U·L^-1;glutamyl oxaloacetic transaminase(GOT)were(95.02±22.10),(186.08±72.36)and(293.64±27.26)U·L^-1;Tbil were(0.86±0.42),(1.82±0.78)and(14.88±7.67)μmol·L^-1;ALB were(34.64±1.90),(27.13±4.26)and(21.55±2.65)g·L^-1;IL^-17 A were(39.84±21.04),(68.24±13.01)and(238.11±32.80)pg·mL^-1;IL-6 were(69.23±14.24),(91.84±5.62)and(99.09±22.44)pg·mL^-1;IL^-10 were(53.83±9.14),(81.67±15.26)and(99.38±25.98)pg·mL^-1;TGF-β1 were(105.86±24.98),(161.86±29.89)and(219.73±59.68)pg·mL^-1;Th17 cell frequency in peripheral blood of control group,test-1 and test-2 groups were(1.33±0.20)%,(5.55±1.57)%and(3.06±0.56)%.Treg cell frequency were(2.53±0.42)%,(4.96±0.61)%and(12.05±1.00)%;The ratio of Th17/Treg was0.52±0.31,1.12±0.30 and 0.25±0.04;The expression of Foxp3 protein(IOD value)in liver tissue of control group,test-1 and test-2 groups were(1.67±0.12),(1.36±0.11)and(1.99±0.14),respectively;the difference

关 键 词：刀豆蛋白A α-半乳糖酰基鞘氨醇 辅助性T细胞17 调节性T细胞 自身免疫性肝炎 

分 类 号：R97[医药卫生—药品]