树突状细胞芳香烃受体诱导辅助性T细胞17分化在原发性胆汁胆管炎中的作用研究  

Study on the role of Aryl hydrocarbon receptor induced Th17 differentiation in dendritic cells in primary biliary cholangitis

作　　者：佘春晖 王静[1] 刘斌[1] She Chunhui;Wang Jing;Liu Bin(Department of Rheumatology,the Affiliated Hospital of Qingdao University,Shandong 266000,China)

机构地区：[1]青岛大学附属医院风湿免疫科,266000

出　　处：《中华风湿病学杂志》2020年第7期433-436,共4页Chinese Journal of Rheumatology

基　　金：国家自然科学基金(81671600);山东省自然科学基金(ZR2016HM13)。

摘　　要：目的观察芳香烃受体(AhR)激活后的树突状细胞(DC)对原发性胆汁性胆管炎(PBC)患者辅助性T细胞17(Th17)分化的影响。方法收集10例PBC患者和10名健康者的外周血标本,分选出CD14+单核细胞,诱导其成为DCs,被2,3,7,8-四氯二苯并对二英激活后与初始CD4+T细胞共培养。然后采用CCK-8法检测T细胞的增殖活性,实时荧光定量PCR(qRT-PCR)与ELISA检测Th17分化相关因子变化。采用t检验,单因素方差分析进行统计学分析。结果获得大量DCs,显微镜下观察可见细胞体积增大,形态不规则,表面大量突起,CD11和CD14双阳性的细胞占96.48%。共培养后,PBC患者组初始CD4+T细胞的增殖能力高于健康对照组,CCK-8法72 h测得的吸光度(A)值(1.02±0.04),较24 h(0.69±0.04)和48 h(0.82±0.04)时的A值增高,差异具有统计学意义(LSD-t1=10.2,P<0.05;LSD-t2=6.3,P<0.05);PBC患者组初始CD4+T细胞IL-17的相对表达量(1.24±0.40)高于健康对照组(0.61±0.18),差异具有统计学意义(t=3.90,P<0.05);IL-22表达量(0.96±0.46)同样高于健康对照组(0.42±0.43),差异具有统计学意义(t=2.76,P<0.05);PBC患者组培养上清液中IL-22的表达量为(77.4±2.9)ng/ml,高于健康对照组(49.6±4.0)ng/ml,差异具有统计学意义(t=5.58,P<0.01);IL-17表达量为(170.9±2.7)ng/ml,同样高于健康对照组(130.6±7.1)ng/ml,差异具有统计学意义(t=9.10,P<0.05)。结论AhR激活后的DCs可以影响Th17的增殖和分化,并诱导其分泌Th17细胞相关的炎性因子,因此AhR激活后的DCs在PBC的发病中可能促进作用。Objective To explore the effect of Aryl hydrocarbon receptor(AhR)activated dendritic cell(DC)on T helper cell 17(Th17)differentiation in primary biliary cholangitis(PBC)patients.Methods Per-ipheral blood samples of 10 patients with PBC and 10 healthy people were collected.CD14+mononuclear cells were isolated and induced to differentiate to DCs.After stimulated by 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD),these DCs were co-cultured with the naïve CD4+T cells.Then,the proliferation activity of T cells was detected by Cell Counting Kit-8(CCK-8)and cytokines related to Th17 differentiation were detected by quantitative real-time-polymerase chain reaction(qRT-PCR)and enzyme-linked immuno sorbent assay(ELISA).Results A large number of DCs were obtained,and the microscopic observation showed that the cells were enlarged in size,irregular in shape,and numerous protrusions on the surface.The CD11 and CD14 double positive cells accounted for 96.48%.After co-culture,the proliferation capacity of naïve CD4+T cells in the PBC group was significantly higher than that in the healthy control group.Absorbance(A)value measured by CCK-8 method at 72 hours(1.02±0.04)was significantly higher than that at 24 hours(0.69±0.04)and 48 hours(0.82±0.04),and the difference was statistically significant(LSD-t1=10.2,P<0.05;LSD-t2=6.3,P<0.05).The expression level of naïve CD4+T cells IL-17 in the PBC patients group was(1.24±0.40)higher than that in the healthy control group(0.61±0.18),and the difference was statistically significant(t=3.90,P<0.05).The expression level of IL-22(0.96±0.46)was also higher than that of the healthy control group(0.42±0.43),and the difference was statistically significant(t=2.76,P<0.05).The expression of IL-22 in the culture supernatant of the PBC patients group was(77.4±2.9)ng/ml,significantly higher than that of the healthy control group(49.6±4.0)ng/ml,the difference was statistically significant(t=5.58,P<0.01).The expression level of IL-17(170.9±2.7)ng/ml was also higher than that of the healthy contr

关 键 词：胆汁性胆管炎 原发性 芳香烃受体 树突状细胞 T淋巴细胞 

分 类 号：R575.7[医药卫生—消化系统]