摘要
目的探究麻黄细辛附子汤对Th2偏移、Treg细胞功能及Notch信号通路的干预作用。方法体外培养CD4+T细胞,流式细胞仪鉴定纯度,使用白细胞介素-4(IL-4)、干扰素-γ抗体(Anti-IFN-γ)诱导建立Th2细胞分化模型,DAPT及麻黄细辛附子汤干预24 h后收集上清及细胞,ELISA检测白细胞介素-5(IL-5)、干扰素-γ(IFN-γ)、转化生长因子β1(TGF-β1)含量,RT-PCR检测T细胞特异性转录因子(T-bet)、转录因子结合蛋白-3(GATA-3)、叉头翼状螺旋转录因子3(FOXP3)、Notch1 mRNA表达,Western blotting检测T-bet、GATA-3、FOXP3、Notch1蛋白表达。结果1)较之空白组,模型组IL-5含量升高,IFN-γ、TGF-β1含量降低(P<0.01);较之模型组,各给药组IL-5含量降低,IFN-γ、TGF-β1含量升高(P<0.01)。2)较之空白组,模型组Tbet、FOXP3 mRNA表达降低,GATA-3、Notch1 m RNA表达升高(P<0.01);较之模型组,各给药组T-bet、FOXP3 mRNA表达升高,GATA-3、Notch1 m RNA表达降低(P<0.01)。3)较之空白组,模型组T-bet、FOXP3蛋白表达降低,GATA-3、Notch1蛋白表达升高(P<0.01);较之模型组,各给药组T-bet、FOXP3蛋白表达升高,GATA-3、Notch1蛋白表达降低(P<0.01)。结论1)麻黄细辛附子汤能够下调IL-5分泌和GATA-3表达,促进IFN-γ分泌和T-bet表达,纠正Th2偏移。2)麻黄细辛附子汤有助于Treg细胞分化增殖,能够促进TGF-β1分泌和FOXP3表达。3)麻黄细辛附子汤能够通过抑制Notch1表达,促进Treg细胞功能发挥,纠正Th2偏移。
Objective:To explore the intervention effect of Mahuang Xixin Fuzi Decoction on Th2 shift,Treg cell function and Notch signaling pathway.Methods:The CD4+T cells were cultured in vitro,and the cell purity was identified by flow cytometry.The Th2 cell differentiation model was established by induction with IL-4 and AntiIFN-γ.The supernatant and cells were collected after DAPT and Mahuang Xixin Fuzi Decoction intervened for 24hours.ELISA was used to detect the IL-5,IFN-γand TGF-β1 content.The expressions of T-bet,GATA-3,FOXP3and Notch1 mRNA were detected by RT-PCR,and the expressions of T-bet,GATA-3 and FOXP3 proteins were detected by Western blotting.Results:1)Compared with the blank group,IL-5 content was increased and IFN-γand TGF-β1 content was decreased in the model group(P<0.01).Compared with the model group,IL-5 content was decreased and IFN-γand TGF-β1 content was increased in each administration group(P<0.01).2)Compared with the blank group,the expression of T-bet and FOXP3 m RNA was decreased and the expression of GATA-3 and Notch1 mRNA was increased in the model group(P<0.01).Compared with the model group,the expression of T-bet and FOXP3 mRNA was increased and the expression of GATA-3 and Notch1 mRNA was decreased in each administration group(P<0.01).3)Compared with the blank group,T-bet and FOXP3 protein expression was decreased and GATA-3 and Notch1 protein expression was increased in the model group(P<0.01).Compared with the model group,T-bet and FOXP3 protein expression was increased and GATA-3 and Notch1 protein expression was decreased in each administration group(P<0.01).Conclusion:1)Mahuang Xixin Fuzi Decoction can down-regulate IL-5 secretion and GATA-3 expression,and promote IFN-γsecretion and T-bet expression to correct the Th2 shift.2)Mahuang Xixin Fuzi Decoction helps the differentiation and proliferation of Treg cells,and can promote TGF-β1 secretion and FOXP3 expression.3)Mahuang Xixin Fuzi Decoction can promote Treg cell function and correct Th2 shift by inhibiting Notch1 expression.
作者
倪钰莹
孙彤彤
范淑月
吴璐蔚
高婧
崔颖
刘敏
Ni Yuying;Sun Tongtong;Fan Shuyue;Wu Luwei;Gao Jing;Cui Ying;Liu Min(Beijing University of Chinese Medicine,Beijing 100029,China)
出处
《中国中医急症》
2023年第2期200-203,216,共5页
Journal of Emergency in Traditional Chinese Medicine
基金
国家自然科学基金项目(81774375)。
作者简介
通信作者:刘敏,电子邮箱:liumin78@126.com。
