疏肝平喘方通过转化生长因子-β_(1)/Smad信号通路对哮喘小鼠免疫平衡的影响  

Effects of Shugan Pingchuan Formula on Immune Balance in Asthmatic Mice Through TGF-β_(1)/Smad Signaling Pathway

作　　者：顾静雯 朴香 傅伟 程克文 董晨洁 沈毅韵[3] GU Jingwen;PU Xiang;FU Wei;CHENG Kewen;DONG Chenjie;SHEN Yiyun(Baoshan Renhe Hospital,Shanghai 200431,China;Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Baoshan Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201999,China)

机构地区：[1]上海市宝山区仁和医院,上海200431 [2]上海中医药大学,上海201203 [3]上海中医药大学附属宝山医院,上海201999

出　　处：《世界中医药》2023年第14期1974-1978,1985,共6页World Chinese Medicine

基　　金：国家自然科学基金青年科学基金项目(821049251);上海市宝山区仁和医院优秀中青年拔尖人才培养计划项目(BSRHYX-2021-01)。

摘　　要：目的:阐明疏肝平喘方对哮喘小鼠维甲酸相关核孤儿受体γt(RORγt)/叉头框蛋白P3(Foxp3)以及辅助性T细胞17(Th17)/调节性T细胞(Treg)平衡的干预作用,对转化生长因子-β_(1)(TGF-β_(1))/Smad信号通路的影响。方法:建立空白对照组、哮喘模型组、疏肝平喘组及地塞米松组实验动物模型。酶联免疫吸附试验检测肺组织匀浆中白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、白细胞介素-17A(IL-17A)和TGF-β_(1)水平,流式细胞仪检测肺组织中Th17/Treg细胞量,蛋白质印迹法检测肺组织中RORγt、Foxp3的蛋白表达,以及逆转录-聚合酶链反应(RT-PCR)检测肺组织TGF-β_(1)、Smad2、Smad3、Smad7的mRNA表达水平。结果:疏肝平喘方能够降低哮喘小鼠肺组织IL-6、TGF-β_(1),与地塞米松比较,差异均有统计学意义(均P<0.05),调节IL-10、IL-17A,与地塞米松组相当(均P>0.05);Th17/Treg细胞量,RORγt、Foxp3的蛋白表达量,与地塞米松相当(均P>0.05)。与哮喘模型组比较,疏肝平喘方组、地塞米松组的TGF-β_(1)、Smad2、Smad3的mRNA表达水平下降,而Smad7的mRNA表达水平升高,差异均有统计学意义(均P<0.05)。结论:疏肝平喘方在改善气道炎症方面,和地塞米松疗效相当,是通过TGF-β_(1)/Smad信号通路,从而调节哮喘小鼠的Th17/Treg免疫平衡。Objective:To elucidate the intervention effects of Shugan Pingchuan Formula(SGPC)on the balance of retinoic acid-related orphan receptor gamma t(RORγt)/forkhead box protein 3(Foxp3)and helper T cell 17(Th17)/regulatory T cell(Treg)in asthmatic mice and its impact on the transforming growth factor-β_(1)(TGF-β_(1))/Smad signaling pathway.Methods:An experimental animal model was established and mice were divided into a blank control group,an asthma model group,an SGPC group,and a dexamethasone group.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of interleukin-6(IL-6),interleukin-10(IL-10),interleukin-17A(IL-17A),and TGF-β_(1) in lung tissue homogenates.Flow cytometry was used to measure Th17/Treg cell counts in lung tissues.Western blot was performed to determine the protein expression of RORγt and Foxp3 in lung tissues.Reverse transcription-polymerase chain reaction(RT-PCR)was used to measure the mRNA expression levels of TGF-β_(1),Smad2,Smad3,and Smad7 in lung tissues.Results:SGPC could reduce the levels of IL-6 and TGF-β_(1) in lung tissues of asthmatic mice,showing statistically significant differences compared with the dexamethasone group(both P<0.05).In terms of the regulation of IL-10 and IL-17A,the SGPC group was equivalent to the dexamethasone group(both P>0.05).In terms of Th17/Treg cell counts and the protein expression of RORγt and Foxp3,the SGPC group was comparable to the dexamethasone group(all P>0.05).Compared with the asthma model group,the SGPC group and the dexamethasone group showed decreased mRNA expression levels of TGF-β_(1),Smad2,and Smad3 and increased mRNA expression level of Smad7 in lung tissues(all P<0.05).Conclusion:SGPC exhibits comparable efficacy to dexamethasone in improving airway inflammation and achieves this effect by regulating the Th17/Treg immune balance in asthmatic mice through the TGF-β_(1)/Smad signaling pathway.

关 键 词：哮喘 转化生长因子-β_(1)/Smad信号通路 辅助性T细胞17/调节性T细胞免疫平衡 疏肝平喘方 维甲酸相关核孤儿受体γt 叉头框蛋白P3 气道炎症 

分 类 号：R284[医药卫生—中药学] R289.5[医药卫生—中医学]