Th17、Treg细胞失衡与哮喘-慢阻肺重叠综合征的相关性及金匮肾气丸干预后的影响研究  

Study on the correlation between ACOS and Th17/Treg cells' imbalance and on the influence of Jinkui Shenqi pills' intervention

作　　者：肖钦文[1] 李海梅 曾珠[1] 岳国龙 陈献 张诗竹 涂翔[3] 李晓康[3] 何成诗[3] Xiao Qinwen;Li Haimei;Zeng Zhu;Yue Guolong;Chen Xian;Zhang Shizhu;Tu Xiang;Li Xiaokang;He Chengshi(Chengdu University of TCM, Chengdu 610075;Affiliated Hospital of Armed Police Logistics College,Tianjin 300162;Affiliated Hospital of Chengdu University of TCM, Chengdu 610075)

机构地区：[1]成都中医药大学,成都610075 [2]武警后勤学院附属医院,天津300162 [3]成都中医药大学附属医院,成都610075

出　　处：《中药药理与临床》2018年第2期5-9,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：四川省科技计划项目基金(2016JY0008)

摘　　要：目的:明确支气管哮喘-慢性阻塞性肺疾病重叠综合征(ashma-COPD overlap syndrome,ACOS)患者与健康体检者Th17、Treg细胞之间的差异性;明确Th17、Treg细胞与ACOS的关系;明确金匮肾气丸是否对ACOS患者Th17、Treg细胞失衡产生作用。方法:ACOS患者和健康体检者血液标本离心收集血清,Ficoll密度梯度离心法分离外周血单个核细胞(peripheral blood mononuclear cell,PBMC),PBMC体外培养,加入相应抗体后,Th17、Treg细胞表面染色,以CD4+IL-17A+、CD4+CD25+Foxp3+荧光阳性细胞分别标示Th17和Treg细胞,流式细胞仪检测ACOS组和阴性对照组的细胞因子表达,将分离的人PBMC细胞接种于包被CD3抗体的培养板,并用2ug/ml的可溶性CD28抗体刺激其活化扩增,根据实验分组在含药血清组分别加入大鼠给药组的5%、10%、15%、20%金匮肾气丸含药血清,空白对照组加入大鼠空白组的血清,刺激培养72h后收集样本进行后续检测。结果:ACOS组CD25+Foxp3标记的细胞比例低于阴性对照组,而CD4+IL-17A+标记的细胞比例明显高于阴性对照组,使用金匮肾气丸干预后,金匮肾气丸含药血清组CD4+IL-17A+标记的细胞比例显著低于空白对照组;CD25+Foxp3标记的细胞比例显著高于空白对照组,与含药血清浓度相关性较大。结论:ACOS组中Th17细胞的表达水平明显高于阴性对照组,Treg细胞的表达水平显著低于阴性对照组,使用金匮肾气丸含药血清干预后,含药血清组Th17细胞因子的表达水平显著低于空白对照组,Treg细胞因子的表达水平显著高于空白对照组,说明金匮肾气丸可纠正Th17、Treg细胞水平失衡。Objective： To clarify the difference between Th17/Treg cells in the ACOS group and the control group,and the relationship between the Th17/Treg cells and ACOS. Methods： Blood samples from ACOS and control groups were centrifuged to collect serum,PBMC was separated,PBMC was cultured in vitro,then added to antibodies,surfaces of Th17/Treg cells were stained. Th17 and Treg cells were expressed by labeled fluorescent positive cells respectively,and then detected by FCM. PBMC cells were separated in microtiter plates were coated with CD3 antibody,activated by CD28 antibody. 5%,10%,15% and 20% serum containing drug were added to experimental groups,samples were collected after 72 hours’ cultivation and then tested. Results： In group ACOS,levels of Th17 and Treg were lower than those in the control group,medicated serum group,the percentage of IL-17 A ＋ positive cells was higher than that in the control group,Foxp3 ＋ single labeled cells and the proportion of CD25＋Foxp3 labeled cells were higher than those in the control group. They were highly correlated with the concentration of serum containing drugs,the difference was statistically significant,indicating that serum levels of Th17 and Treg in the serum containing drugs groups were higher than those in the control group,and the concentration of serum containing drug was significantly correlated with levels of Th17 and Treg. Conclusion： Th17 and Treg cells are negatively correlated with the level of ACOS,and Jinkui Shenqi pills can correct the imbalance of Th17 and Treg cells.

关 键 词：重叠综合征 辅助性T细胞17 金匮肾气丸 调节性T细胞 

分 类 号：R259[医药卫生—中西医结合]