不同归经芳香药对流行性感冒小鼠气道黏膜影响及差异基因表达谱分析  

Effects of different meridian aromatic drugs on airway mucosa of influenza mice and analysis of differential gene expression profiling

作　　者：王雪飞[1] 张燕 韦丽妮 王瑞琪 鲁瑶 施长琪 李立[2] WANG Xuefei;ZHANG Yan;WEI Lini;WANG Ruiqi;LU Yao;SHI Changqi;LI Li(Acupuncture Center,Beijing Hospital of Traditional Chinese Medicine Affiliated to Capital Medical University,Beijing 100010,China;Institute of Basic Research in Clinical Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]首都医科大学附属北京中医医院针灸中心,北京100010 [2]中国中医科学院中医临床基础医学研究所,北京100700

出　　处：《中华中医药杂志》2024年第7期3403-3409,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家重点研发计划(No.2020YFE0205100)。

摘　　要：目的:研究不同归经的芳香药对流行性感冒(以下简称流感)小鼠的气道黏膜和基因表达谱的差异。方法:40只BALB/c小鼠随机分为空白组、模型组、藿香组及苍术组,每组10只。除空白组外,滴鼻接种FM1病毒制造流感模型,治疗组小鼠分别雾化吸入藿香或苍术,干预5d后处死,留取标本检测。采用HE染色观察肺及气管组织病理形态,ELISA法检测小鼠肺泡灌洗液中分泌型免疫球蛋白A(sIgA)、肿瘤坏死因子-α(TNF-α)、干扰素-γ(IFN-γ)和白介素-1β(IL-1β)表达,流式细胞术检测T淋巴细胞亚群,转录组测序分析小鼠肺组织差异基因,生物信息学技术进行可视化展示。结果:藿香组和苍术组均可减轻流感小鼠肺组织炎症病理损伤,与模型组比较,两给药组均可显著升高sIgA表达(P<0.05),显著降低TNF-α、IFN-γ和IL-1β表达(苍术组IL-1β除外)(P<0.01,P<0.05),显著升高CD4^(+)T淋巴细胞及CD4^(+)/CD8^(+)比值和降低CD8^(+)T淋巴细胞水平(P<0.01,P<0.05)。与苍术组比较,藿香组TNF-α、IFN-γ显著降低(P<0.01),气管黏膜CD4^(+)T、CD8^(+)T淋巴细胞均显著升高(P<0.01)。转录组测序得到模型组、藿香组、苍术组的差异基因分别有1103、311、315个。藿香组、苍术组与模型组共同作用的通路不同。对于白介素10通路,藿香组表现出激活作用,模型组表现出抑制作用,MHC、CREB、BMP2为其共同作用于该条通路的分子,且调节趋势相反。对巨噬细胞选择性激活信号通路,苍术组表现出一定的抑制作用,模型组表现出激活作用,IGG、CREB为其作用于该通路的共同分子,且调节趋势相反。结论:归肺经芳香药藿香较其他归经芳香药苍术对流感呼吸道黏膜免疫调节及炎症控制有一定优势,可为中药归经理论提供参考。Objective:To study the differences in airway mucosa and gene expression profiles of influenza mice by aromatic drugs with lung meridian or not.Methods:Forty BALB/c mice were randomly divided into blank group,model group,Patchouli group and Atractylodes group,10 mice in each group.Except for the blank group,FM1 virus was administered intranasally to establish an influenza model.The treatment group mice were nebulized with Patchouli or Atractylodes,and euthanized after 5 days of intervention.Samples were collected and tested.HE staining was used to observe the pathological morphology of lung and tracheal tissues,ELISA method was for detecting sIgA,TNF-α,IFN-γand IL-1βin mouse alveolar lavage fluid,flow cytometry method was for detecting T lymphocyte,transcriptome sequencing analysis was for detecting differentially expressed genes in mouse lung tissue,and visualization display by using bioinformatics technology.Results:Both Patchouli and Atractylodes could alleviate inflammatory pathological damage to lung tissues in influenza mice.Compared with the model group,both Patchouli and Atractylodes group increased sgA(P<0.05),decreased TNF-α,IFN-γ and IL-1βexpression(except IL-1βexpression in Atractylodes group)(P<0.01,P<0.05),elevated CD4^(+)T,CD4^(+)/CD8^(+)and decreased CD8^(+)T lymphocytes(P<0.01,P<0.05).Compared with Atractylodes group,the decrease of TNF-α,IFN-γ were more pronounced in Patchouli group(P<0.01),the CD4t,CD8^(+)T lymphocytes in the tracheal mucosa were significantly increased(P<0.01).There were 1103,311,and 315 differential genes in the model group,Patchouli group,and Atractylodes groups,respectively.The common pathways were different between in the three groups.For the IL-10 signaling pathway,the Patchouli group showed an activation effect,while the model group showed an inhibitory effect,MHC,CREB and BMP2 were the common molecules in the Patchouli group and model group.In the macrophage alternative activation signaling pathway,the Atractylodes group showed an inhibitory effect,while the mod

关 键 词：流行性感冒 归经 藿香 苍术 黏膜免疫 转录组测序 炎症 基因表达谱 

分 类 号：R285.5[医药卫生—中药学]