益气温阳护卫汤含药血清对体外Th17/Treg细胞失衡与RORγ、Foxp3的影响  

Effects of Yiqi Wenyang Huwei Decoction medicated serum on Th17/Treg cell imbalance invitroand RORy and Foxp3

作　　者：余涛[1] 黄鑫[1] 喻强强[2] 孙朋[2] 向双娣 薛汉荣[1,2] YU Tao;HUANG Xin;YU Qiang-qiang;SUN Peng;XIANG Shuang-di;XUE Han-rong(Jiangxi University of Chinese Medicine,Nanchang 330004,China;Affliated Hospital of Jiangxi University of Chinese Medicine,Nanchang 330006,China)

机构地区：[1]江西中医药大学,南昌330004 [2]江西中医药大学附属医院,南昌330006

出　　处：《中华中医药杂志》2023年第4期1554-1558,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81960802,No.81760801);江西中医药大学博士科研启动基金项目(No.2021BSZRO27)。

摘　　要：目的:研究益气温阳护卫汤含药血清对体外辅助性T淋巴细胞17(Th17)/调节性T细胞(Treg)细胞失衡与维甲酸相关孤核受体γ(RORγ)、特异性叉头状转录因子3(Foxp3)的影响。方法:制备益气温阳护卫汤含药血清,CD4+T细胞进行原代分离、培养、鉴定。构建体外Th17/Treg细胞失衡模型:将细胞分为对照组、模型组、益气温阳护卫组、地塞米松组,除对照组外,其余组中分别加入等量转化生长因子-β(TGF-β)、白细胞介素(IL)-6、IL-23溶液,诱导CD4+T细胞向Th17细胞分化。给予相应的含药血清干预处理48 h后,采用流式细胞术检测各组Th17和Treg细胞比例。RT-PCR、Western Blot分别检测RORγ、Foxp3 mRNA和蛋白表达水平。结果:与对照组比较,模型组Th17细胞比例、Th17/Treg比例均显著上升(P<0.01),Treg细胞比例显著下降(P<0.01);与模型组比较,益气温阳护卫组及地塞米松组Th17细胞比例、Th17/Treg比例均显著降低(P<0.01,P<0.05),Treg细胞比例显著上升(P<0.01,P<0.05)。与对照组比较,模型组RORγ mRNA及蛋白表达均显著升高(P<0.05,P<0.01),Foxp3 mRNA及蛋白表达均显著降低(P<0.01,P<0.05);与模型组比较,益气温阳护卫组及地塞米松组RORγ mRNA及蛋白表达均显著降低(P<0.05),Foxp3 mRNA及蛋白表达均显著升高(P<0.05,P<0.01)。结论:益气温阳护卫汤含药血清能有效改善Th17/Treg细胞体外失衡,其机制可能与调节特异性转录因子RORγ、Foxp3的表达相关。Objective:To study the effect of Yiqi Wenyang Huwei Decoction medicated serum on Th17/Treg cell imbalance in vitro and ROR and Foxp3.Methods:The serum containing Yiqi Wenyang Huwei Decoction was prepared,and CD,T cells were isolated,cultured and identified.To construct the imbalance model of Th17/Treg cells in vitro:the cells were divided into control group,model group,Yiqi Wenyang Huwei group and dexamethasone group.In addition to the control group,the other groups were added with the same amount of TGF-β,IL-6 and IL-23 solution which induced CD&T cells to differentiate into Th17 cells.After 48 hours of treatment,the proportion of Th17 and Treg cells in each group was detected by flow cytometry.ROR,Foxp3 mRNA and protein expression levels were detected by RT-PCR and Western Blot.Results:Compared with the control group,the proportion of Th17 cells and Th17/Treg in the model group significantly increased(P<0.01),and the proportion of Treg cells significantly decreased(P<0.01).Compared with the model group,the proportion of Th17 cells and Th17/Treg in the Yiqi Wenyang Huwei group and dexamethasone group significantly decreased(P<0.01,P<0.05),and the proportion of Treg cells significantly increased(P<0.01,P<0.05).Compared with the control group,the expression of RORy mRNA and protein the of the model group significantly increased(P<0.05,P<0.01),and the expression of Foxp3 mRNA and protein significantly decreased(P<0.01,P<0.05).Compared with the model group,the expression of ROR mRNA and protein of Yiqi Wenyang Huwei group and dexamethasone group significantly decreased(P<0.05),and the expression of Foxp3 mRNA and protein significantly increased(P<0.05,P<0.01).Conclusion:Yiqi Wenyang Huwei Decoction medicated serum can effectively improve the imbalance of Th17/Treg cells in vitro,and its mechanism may be related to the regulation of specific transcription factor RORy,Foxp3 expression.

关 键 词：益气温阳护卫汤 哮喘 Th17/Treg细胞失衡 维甲酸相关孤核受体γ 特异性叉头状转录因子3 机制 

分 类 号：R285.5[医药卫生—中药学]