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Article(id=1322927063807701685, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322926985957221241, articleNumber=null, orderNo=null, doi=10.13193/j.issn.1673-7717.2025.11.015, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=null, receivedDateStr=null, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1793339990807, onlineDateStr=2026-10-30, pubDate=1762704000000, pubDateStr=2025-11-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793339990807, onlineIssueDateStr=2026-10-30, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793339990807, creator=13701087609, updateTime=1793339990807, updator=13701087609, issue=Issue{id=1322926985957221241, tenantId=1146029695717560320, journalId=1255847919539208197, year='2025', volume='43', issue='11', pageStart='1', pageEnd='258', issueExtLink='null', onlineDate='null', pubDate='1762704000000', pubDateStr='2025-11-10', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1793339972247, creator='13701087609', updateTime=1793340743335, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1322930220201804556, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322926985957221241, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1322930220201804557, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322926985957221241, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=64, endPage=68, ext={EN=ArticleExt(id=1322927063988056758, articleId=1322927063807701685, tenantId=1146029695717560320, journalId=1255847919539208197, language=EN, title=Immunological Mechanism of Shegan Mahuang Decoction（射干麻黄汤）in Modulating T Lymphocytes to Improve Airway Hyperresponsiveness in Mice through Simultaneous Treatment of Nose and Lung, columnId=1256183361521967297, journalTitle=Chinese Archives of Traditional Chinese Medicine, columnName=Target on National Project, runingTitle=null, highlight=null, articleAbstract=
Objective

To observe the effect of Shegan Mahuang Decoction（射干麻黄汤）on T lymphocyte subsets in upper and lower airways of airway hyperresponsiveness（AHR）mice，and analyze the biological basis of simultaneous treatment of nose and lung in AHR treatment.

Methods

Gammadelta（γδ）T cells were purified from beta T receptor knockout mice. AHR model mice sensitized with ovalbumin were randomly attributed into normal control group（Ctrl），AHRgroup（AHR），γδT cells adoptive immunization group（AIT），Shegan Mahuang Decoction group（SGMHD），Shegan Mahuang Decoction and adoptive immunization group（SMA）. AIT group was injectedγδT cells via tail vein daily，SGMHTgroup was given Shegan Mahuang Decoction daily by gavage，and SMA group was given dailyγδT cell tail vein injection combined with Shegan Mahuang Decoction by gavage for 7 days. T lymphocyte subsetsmarkers in nasal mucosa and lung of mice were detected，including interferon-γ（IFN-γ），interleukin-4（IL-4），retinoic acid related orphan receptorγt（RORγt），forkhead transcription factor P3（Fox P3）andγδTcell receptor（γδTCR）. The ratios of Th1，Th2，Th17，Treg andγδT cells，and the corresponding cytokines in peripheral blood were also measured.

Results

The results showed that compared with those in the Ctrl group，the expressions of IFN-γ，IL-4，FoxP3，RORγt andγδTCR of nasal mucosa and lung were higher in AHR group，with higher proportions of Th2，Th17 andγδT cells，higher concentrations of IL-2，IL-4，IL-5 and IL-17，and lower concentrations of IL-10 and TGF-β. Compared with those in the AHR group，the expressions of IFN-γ，IL-4，FoxP3，RORγt andγδTCRof nasal mucosa were lower in the AIT，SGMHD and SMA groups，with lower levels of IL-4 andγδTCR in lung，higher ratio of Th1 and lower ratios of Th17 andγδT cells in peripheral blood. The levels of IL-2，IL-4，IL-5 and IL-17 of SGMHT and SMA groups were lower expressed，however the levels of IL-10 and TGF-βwere higher.

Conclusion

The study clarified the effect of Shegan Mahuang Decoction on AHR mice through simultaneous treatment of nose and lung. The underlying mechanism was related to the comprehensive regulation of T lymphocyte subsets. The study laid a foundation for revealing mechanisms of Shegan Mahuang Decoction in the treatment of AHR and promoting its clinical practice.

, authors=Jingyi ZHAO, Xinyu YAN, Man WANG, Suying GUO, Hanyi DONG, Junge WANG, authorsList=Jingyi ZHAO, Xinyu YAN, Man WANG, Suying GUO, Hanyi DONG, Junge WANG, authorCompany=null, correspAuthors=Junge WANG, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1322927065095353023, articleId=1322927063807701685, tenantId=1146029695717560320, journalId=1255847919539208197, language=CN, title=射干麻黄汤“鼻肺同治”调控T淋巴细胞改善小鼠气道高反应的免疫机制, columnId=1256183362172142382, journalTitle=中华中医药学刊, columnName=国家项目点击, runingTitle=null, highlight=null, articleAbstract=
目的

观察射干麻黄汤对气道高反应（AHR）小鼠模型上、下气道T淋巴细胞亚群的影响，分析其“鼻肺同治”控制AHR的生物学基础。

方法

采用T细胞β受体基因敲除小鼠提纯γδT细胞；采用卵清蛋白气道致敏的AHR模型小鼠，平均随机分为正常对照组（Ctrl组）、气道高反应模型组（AHR组）、过继免疫γδT细胞组（AIT组）、射干麻黄汤干预组（SGMHD组）、射干麻黄汤＋过继免疫组（SMA组），AIT组予以每日γδT细胞尾静脉注射，SGMHD组予以每日射干麻黄汤灌胃，SMA组予以每日γδT细胞尾静脉注射联合射干麻黄汤灌胃，干预7 d。观察各组小鼠鼻黏膜和肺组织中T淋巴细胞亚群标志物干扰素-γ（IFN-γ）、白细胞介素-4（IL-4）、视黄酸相关孤儿受体γt（RORγt）、叉头状转录因子P3（FoxP3）及γδT细胞受体（γδTCR）分布情况，外周血Th1、Th2、Th17、Treg、γδT细胞及其细胞因子的表达水平。

结果

与Ctrl组相比，AHR组鼻黏膜和肺组织中IFN-γ、IL-4、FoxP3、RORγt、γδTCR表达较高，外周血Th2、Th17、γδT细胞比例较高，IFN-γ、IL-2、IL-4、IL-5及IL-17浓度均较高，IL-10、TGF-β则表达较低；与AHR组相比，AIT组、SGMHD组及SMA组鼻黏膜IFN-γ、IL-4、RORγt、γδTCR表达较低，肺组织IL-4、γδTCR较低，外周血Th1比例较高，Th17、γδT较低，SGMHD组和SMA组IFN-γ、IL-2、IL-4、IL-5及IL-17浓度较低，IL-10、TGF-β较高。

结论

明确了射干麻黄汤“鼻肺同治”改善AHR模型小鼠的作用，其机制与全面调控T淋巴细胞亚群有关。研究为揭示射干麻黄汤治疗AHR作用机制、促进临床应用奠定基础。

, authors=赵竞一, 闫新宇, 王曼, 郭苏影, 董寒毅, 王俊阁, authorsList=赵竞一, 闫新宇, 王曼, 郭苏影, 董寒毅, 王俊阁, authorCompany=null, correspAuthors=王俊阁, authorNote=

赵竞一（1989-），男，山东潍坊人，主治医师，硕士，研究方向：经方治疗气道变态反应性疾病。

, correspAuthorsNote=
王俊阁（1967-），男，河北沧州人，主任医师、教授，博士研究生导师，博士，研究方向：经方治疗气道变态反应性疾病。E-mail：。
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赵竞一（1989-），男，山东潍坊人，主治医师，硕士，研究方向：经方治疗气道变态反应性疾病。

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赵竞一（1989-），男，山东潍坊人，主治医师，硕士，研究方向：经方治疗气道变态反应性疾病。

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组别IFN-γIL-4FoxP3RORγγδTCR
Ctrl组0.126±0.0240.151±0.0360.140±0.0210.132±0.0250.101±0.022
AHR组0.308±0.053***0.323±0.048***0.208±0.033**0.294±0.030***0.232±0.035***
AIT组0.229±0.052#0.185±0.031###0.153±0.039#0.160±0.019###0.140±0.030###
SGMHT组0.157±0.058###0.172±0.025###0.213±0.0470.161±0.018###0.143±0.040###
SMA组0.173±0.072###0.216±0.045##0.207±0.0430.157±0.031###0.141±0.052###
F值14.027.06.853.014.0
P值<0.001<0.001<0.001<0.001<0.001
), ArticleFig(id=1322927069449040620, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322927063807701685, language=CN, label=表1, caption=

各组小鼠鼻黏膜T淋巴细胞亚群蛋白免疫组化平均光密度值（±s）

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组别IFN-γIL-4FoxP3RORγγδTCR
Ctrl组0.126±0.0240.151±0.0360.140±0.0210.132±0.0250.101±0.022
AHR组0.308±0.053***0.323±0.048***0.208±0.033**0.294±0.030***0.232±0.035***
AIT组0.229±0.052#0.185±0.031###0.153±0.039#0.160±0.019###0.140±0.030###
SGMHT组0.157±0.058###0.172±0.025###0.213±0.0470.161±0.018###0.143±0.040###
SMA组0.173±0.072###0.216±0.045##0.207±0.0430.157±0.031###0.141±0.052###
F值14.027.06.853.014.0
P值<0.001<0.001<0.001<0.001<0.001
), ArticleFig(id=1322927069511955181, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322927063807701685, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别IFN-γIL-4FoxP3RORγγδTCR
Ctrl组0.026±0.0050.023±0.0060.169±0.0220.119±0.0290.085±0.024
AHR组0.314±0.058***0.246±0.048***0.341±0.056***0.301±0.037***0.272±0.051***
AIT组0.232±0.049b#0.170±0.040###0.353±0.0320.281±0.0310.138±0.024###
SGMHT组0.343±0.0630.123±0.020###0.181±0.023###0.300±0.0370.165±0.035###
SMA组0.357±0.0650.125±0.022###0.175±0.023###0.291±0.0430.161±0.041###
F值54.055.062.039.028.0
P值<0.001<0.001<0.001<0.001<0.001
), ArticleFig(id=1322927069579064046, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322927063807701685, language=CN, label=表2, caption=

各组小鼠肺组织T淋巴细胞亚群蛋白免疫组化平均光密度值（±s）

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组别IFN-γIL-4FoxP3RORγγδTCR
Ctrl组0.026±0.0050.023±0.0060.169±0.0220.119±0.0290.085±0.024
AHR组0.314±0.058***0.246±0.048***0.341±0.056***0.301±0.037***0.272±0.051***
AIT组0.232±0.049b#0.170±0.040###0.353±0.0320.281±0.0310.138±0.024###
SGMHT组0.343±0.0630.123±0.020###0.181±0.023###0.300±0.0370.165±0.035###
SMA组0.357±0.0650.125±0.022###0.175±0.023###0.291±0.0430.161±0.041###
F值54.055.062.039.028.0
P值<0.001<0.001<0.001<0.001<0.001
), ArticleFig(id=1322927069646172911, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322927063807701685, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别Th1Th2Th17TregγδT
Ctrl组3.21±0.870.67±0.301.35±0.4717.34±1.103.52±0.43
AHR组3.33±0.591.71±0.30***4.09±1.17***15.87±1.145.58±0.78***
AIT组8.20±1.59###1.65±0.381.55±0.30###14.04±1.27b#3.60±0.53###
SGMHT组5.27±0.84##1.62±0.251.36±0.46###14.82±1.424.24±0.94##
SMA组5.45±0.85##1.47±0.281.30±0.25###16.99±0.964.21±0.60##
F值33.016.030.011.012.0
P值<0.001<0.001<0.001<0.001<0.001
), ArticleFig(id=1322927069721670384, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322927063807701685, language=CN, label=表3, caption=

各组小鼠不同T细胞亚群细胞比例（±s）

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组别Th1Th2Th17TregγδT
Ctrl组3.21±0.870.67±0.301.35±0.4717.34±1.103.52±0.43
AHR组3.33±0.591.71±0.30***4.09±1.17***15.87±1.145.58±0.78***
AIT组8.20±1.59###1.65±0.381.55±0.30###14.04±1.27b#3.60±0.53###
SGMHT组5.27±0.84##1.62±0.251.36±0.46###14.82±1.424.24±0.94##
SMA组5.45±0.85##1.47±0.281.30±0.25###16.99±0.964.21±0.60##
F值33.016.030.011.012.0
P值<0.001<0.001<0.001<0.001<0.001
), ArticleFig(id=1322927069797167857, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322927063807701685, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别IFN-γIL-2IL-4IL-5IL-10IL-17TGF-β
Ctrl组80.91±22.5653.19±10.9756.11±10.2475.69±14.02121.70±16.7621.04±4.12225.40±45.86
AHR组120.80±23.70**165.40±36.68***169.40±38.77***195.20±30.18***38.05±11.52***49.90±7.77***33.97±6.52***
AIT组71.37±12.01###120.40±27.35##132.10±30.53#169.00±37.1823.22±7.0831.71±5.20###126.20±26.52###
SGMHT组67.01±16.63###73.78±10.10###70.98±10.65###103.20±20.40###84.38±15.95###21.53±2.38###161.20±26.17###
SMA组67.51±16.70###74.66±13.17###71.78±12.37###104.17±16.34###84.45±12.98###22.32±5.78###162.32±28.30###
F值12.033.034.032.071.042.045.0
P值<0.001<0.001<0.001<0.001<0.001<0.001<0.001
), ArticleFig(id=1322927071479083762, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322927063807701685, language=CN, label=表4, caption=

Elisa法检测各组小鼠血清不同细胞因子浓度结果（±s）

, figureFileSmall=null, figureFileBig=null, tableContent=
组别IFN-γIL-2IL-4IL-5IL-10IL-17TGF-β
Ctrl组80.91±22.5653.19±10.9756.11±10.2475.69±14.02121.70±16.7621.04±4.12225.40±45.86
AHR组120.80±23.70**165.40±36.68***169.40±38.77***195.20±30.18***38.05±11.52***49.90±7.77***33.97±6.52***
AIT组71.37±12.01###120.40±27.35##132.10±30.53#169.00±37.1823.22±7.0831.71±5.20###126.20±26.52###
SGMHT组67.01±16.63###73.78±10.10###70.98±10.65###103.20±20.40###84.38±15.95###21.53±2.38###161.20±26.17###
SMA组67.51±16.70###74.66±13.17###71.78±12.37###104.17±16.34###84.45±12.98###22.32±5.78###162.32±28.30###
F值12.033.034.032.071.042.045.0
P值<0.001<0.001<0.001<0.001<0.001<0.001<0.001
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射干麻黄汤“鼻肺同治”调控T淋巴细胞改善小鼠气道高反应的免疫机制
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赵竞一 , 闫新宇 , 王曼 , 郭苏影 , 董寒毅 , 王俊阁
中华中医药学刊 | 国家项目点击 2025,43(11): 64-68
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中华中医药学刊 |国家项目点击 2025 , 43 (11) : 64 -68
射干麻黄汤“鼻肺同治”调控T淋巴细胞改善小鼠气道高反应的免疫机制
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赵竞一（1989-），男，山东潍坊人，主治医师，硕士，研究方向：经方治疗气道变态反应性疾病。

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赵竞一, 闫新宇, 王曼, 郭苏影, 董寒毅, 王俊阁
作者信息
  • 首都医科大学附属北京中医医院，北京 100010
通讯作者:
王俊阁（1967-），男，河北沧州人，主任医师、教授，博士研究生导师，博士，研究方向：经方治疗气道变态反应性疾病。E-mail：。
作者简介:

赵竞一（1989-），男，山东潍坊人，主治医师，硕士，研究方向：经方治疗气道变态反应性疾病。

Immunological Mechanism of Shegan Mahuang Decoction（射干麻黄汤）in Modulating T Lymphocytes to Improve Airway Hyperresponsiveness in Mice through Simultaneous Treatment of Nose and Lung
Jingyi ZHAO, Xinyu YAN, Man WANG, Suying GUO, Hanyi DONG, Junge WANG
Affiliations
  • Beijing Hospital of Traditional Chinese Medicine，Beijing 100010，China
出版时间: 2025-11-10 doi: 10.13193/j.issn.1673-7717.2025.11.015
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摘要
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目的

观察射干麻黄汤对气道高反应（AHR）小鼠模型上、下气道T淋巴细胞亚群的影响，分析其“鼻肺同治”控制AHR的生物学基础。

方法

采用T细胞β受体基因敲除小鼠提纯γδT细胞；采用卵清蛋白气道致敏的AHR模型小鼠，平均随机分为正常对照组（Ctrl组）、气道高反应模型组（AHR组）、过继免疫γδT细胞组（AIT组）、射干麻黄汤干预组（SGMHD组）、射干麻黄汤＋过继免疫组（SMA组），AIT组予以每日γδT细胞尾静脉注射，SGMHD组予以每日射干麻黄汤灌胃，SMA组予以每日γδT细胞尾静脉注射联合射干麻黄汤灌胃，干预7 d。观察各组小鼠鼻黏膜和肺组织中T淋巴细胞亚群标志物干扰素-γ（IFN-γ）、白细胞介素-4（IL-4）、视黄酸相关孤儿受体γt（RORγt）、叉头状转录因子P3（FoxP3）及γδT细胞受体（γδTCR）分布情况，外周血Th1、Th2、Th17、Treg、γδT细胞及其细胞因子的表达水平。

结果

与Ctrl组相比，AHR组鼻黏膜和肺组织中IFN-γ、IL-4、FoxP3、RORγt、γδTCR表达较高，外周血Th2、Th17、γδT细胞比例较高，IFN-γ、IL-2、IL-4、IL-5及IL-17浓度均较高，IL-10、TGF-β则表达较低；与AHR组相比，AIT组、SGMHD组及SMA组鼻黏膜IFN-γ、IL-4、RORγt、γδTCR表达较低，肺组织IL-4、γδTCR较低，外周血Th1比例较高，Th17、γδT较低，SGMHD组和SMA组IFN-γ、IL-2、IL-4、IL-5及IL-17浓度较低，IL-10、TGF-β较高。

结论

明确了射干麻黄汤“鼻肺同治”改善AHR模型小鼠的作用，其机制与全面调控T淋巴细胞亚群有关。研究为揭示射干麻黄汤治疗AHR作用机制、促进临床应用奠定基础。

射干麻黄汤  /  气道高反应  /  气道炎症  /  T淋巴细胞  /  细胞因子
Objective

To observe the effect of Shegan Mahuang Decoction（射干麻黄汤）on T lymphocyte subsets in upper and lower airways of airway hyperresponsiveness（AHR）mice，and analyze the biological basis of simultaneous treatment of nose and lung in AHR treatment.

Methods

Gammadelta（γδ）T cells were purified from beta T receptor knockout mice. AHR model mice sensitized with ovalbumin were randomly attributed into normal control group（Ctrl），AHRgroup（AHR），γδT cells adoptive immunization group（AIT），Shegan Mahuang Decoction group（SGMHD），Shegan Mahuang Decoction and adoptive immunization group（SMA）. AIT group was injectedγδT cells via tail vein daily，SGMHTgroup was given Shegan Mahuang Decoction daily by gavage，and SMA group was given dailyγδT cell tail vein injection combined with Shegan Mahuang Decoction by gavage for 7 days. T lymphocyte subsetsmarkers in nasal mucosa and lung of mice were detected，including interferon-γ（IFN-γ），interleukin-4（IL-4），retinoic acid related orphan receptorγt（RORγt），forkhead transcription factor P3（Fox P3）andγδTcell receptor（γδTCR）. The ratios of Th1，Th2，Th17，Treg andγδT cells，and the corresponding cytokines in peripheral blood were also measured.

Results

The results showed that compared with those in the Ctrl group，the expressions of IFN-γ，IL-4，FoxP3，RORγt andγδTCR of nasal mucosa and lung were higher in AHR group，with higher proportions of Th2，Th17 andγδT cells，higher concentrations of IL-2，IL-4，IL-5 and IL-17，and lower concentrations of IL-10 and TGF-β. Compared with those in the AHR group，the expressions of IFN-γ，IL-4，FoxP3，RORγt andγδTCRof nasal mucosa were lower in the AIT，SGMHD and SMA groups，with lower levels of IL-4 andγδTCR in lung，higher ratio of Th1 and lower ratios of Th17 andγδT cells in peripheral blood. The levels of IL-2，IL-4，IL-5 and IL-17 of SGMHT and SMA groups were lower expressed，however the levels of IL-10 and TGF-βwere higher.

Conclusion

The study clarified the effect of Shegan Mahuang Decoction on AHR mice through simultaneous treatment of nose and lung. The underlying mechanism was related to the comprehensive regulation of T lymphocyte subsets. The study laid a foundation for revealing mechanisms of Shegan Mahuang Decoction in the treatment of AHR and promoting its clinical practice.

Shegan Mahuang Decoction（射干麻黄汤）  /  airway hyperresponsiveness  /  airway inflammation  /  T lymphocytes  /  cytokines
赵竞一, 闫新宇, 王曼, 郭苏影, 董寒毅, 王俊阁. 射干麻黄汤“鼻肺同治”调控T淋巴细胞改善小鼠气道高反应的免疫机制. 中华中医药学刊, 2025 , 43 (11) : 64 -68 . DOI: 10.13193/j.issn.1673-7717.2025.11.015
Jingyi ZHAO, Xinyu YAN, Man WANG, Suying GUO, Hanyi DONG, Junge WANG. Immunological Mechanism of Shegan Mahuang Decoction（射干麻黄汤）in Modulating T Lymphocytes to Improve Airway Hyperresponsiveness in Mice through Simultaneous Treatment of Nose and Lung[J]. Chinese Archives of Traditional Chinese Medicine, 2025 , 43 (11) : 64 -68 . DOI: 10.13193/j.issn.1673-7717.2025.11.015
正文
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气道高反应（airway hyper reactivity，AHR）是变应性鼻炎、变应性哮喘、急/慢性支气管炎和新型冠状病毒感染等多种气道过敏性、感染性疾病的共同病理生理特征[1-2]，以过敏原或其他病原体刺激时出现气道过度收缩为主要表现，引发咳嗽、喘息及呼吸困难等症状，严重时危及患者生命。AHR常伴有气道重塑等不可逆性改变，可诱发阻塞性肺气肿、肺纤维化等严重呼吸系统疾病[3]，故控制AHR是治疗气道过敏性和感染性疾病的重要环节。AHR发生机制十分复杂，研究发现辅助性T细胞1型（T helper 1，Th1）/辅助性T细胞2型（T helper 2，Th2）、辅助性T细胞17型（T helper 17，Th17）/调节性T细胞（regulatory T cells，Treg）免疫失衡是AHR发病机制中的主要环节[4-5]。我们的前期研究[6-7]表明，贯通固有免疫和适应性免疫进程的γδT细胞对呼吸道黏膜具有重要的免疫监视和维护稳态作用，在呼吸道黏膜受刺激或损伤的情况下，γδT细胞激活并产生多种细胞因子如白细胞介素（interleukin，IL）、干扰素-γ（interferon-γ，IFN-γ）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）及转化生长因子-β（transforming growth factor-β，TGF-β）等，调控Th1/Th2和Treg/Th17免疫反应，促进气道损伤。
中医古籍中虽无关于“气道高反应”的记载，但从其发病特点来看，AHR宜属于“哮证”范畴，病位在肺，与脾肾相关。病因以痰为主，平素痰伏于肺，发时痰气搏结、肺失宣降、壅阻气道，加之脾失健运、继而痰浊内生，上干于肺，发为“哮证”。《金匮要略·肺痿肺痈咳嗽上气病脉证治第七》载：“咳而上气，喉中水鸡声，射干麻黄汤主之。”射干麻黄汤是临床治疗咳喘的常用方，也是治疗AHR的重要方剂[8]。现代药理学研究也表明射干麻黄汤具有解痉平喘、减轻气道炎症反应的作用[9-11]。本研究通过实验观察射干麻黄汤对AHR模型小鼠上、下气道淋巴细胞及其细胞因子的影响，分析射干麻黄汤“鼻肺同治”控制AHR的现代生物学基础。
1　实验材料
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1.1　实验动物
敲除T细胞β受体基因（T cell receptor beta knock out，TCRβ-KO）的雄性C57BL/6小鼠5只，购自上海南方模式生物科技股份有限公司，目录号NM-KO-190437。SPF级6周大小雄性健康C57BL/6小鼠（体质量18～20 g）40只，购自北京维通利华实验动物技术有限公司，试验动物许可证号SYXK（京）2022-0015。所有小鼠在试验前适应性饲养1周，给予自由进食及饮水，温度范围23～26℃，相对湿度50% ～70%，12 h光亮/黑暗循环。
1.2　主要仪器设备
ELx800全自动酶标仪（BioTek）；FACSCalibur流式细胞仪（BD Biosciences）；Benchmark XT全自动免疫组化仪（Roche）；Allegra X-15R离心机（Beckman）；HistoCore PEGASUS组织脱水机、HistoCore Arcadia包埋机、HistoCore BIOCUT切片机（Leica）；ECLIPSE Si正置显微镜（上海尼康精机有限公司）；DHG-9140A鼓风干燥箱（上海一恒公司）。
1.3　药物与试剂
中药饮片射干、细辛、五味子、紫菀、款冬花、姜半夏、大枣购自首都医科大学附属北京中医医院药事部。
小鼠TCRγ/δ＋T细胞分离试剂盒（批号5801254120），德国miltenyi公司；卵清蛋白（OVA，批号85907798），氢氧化铝（批号74525124），美国Sigma公司；IL-4（批号9365301）、IL-10（批号8487472）、IL-17（批号9484202）、IL-23（批号9095944）、TNF-α（批号8910411）、IFN-γ（批号8330300）、TGF-β1（批号8576699）ELISA试剂盒、鼠单抗CD4 PerCPCy5.5（批号17088756）及鼠单抗CD8 PE-Cy7（批号19752857），IL-4 APC（批号27235041）、IFN-γFITC（批号18708171）、IL-17 PE（批号32387275）、FoxP3 PE-Cy5（批号11322232），美国ThermoFisher公司；RPMI1640培养基（批号31800022），GIBCO 公司；佛波酯/莫能霉素混合物（批号212521257），中国联科生物；鼠T淋巴细胞亚群抗体Cocktail（批号7752962），美国BD公司；鼠单抗γδ-TCR抗体（批号1330048），美国ImmunoTech公司；RORγt（批号540964）、FoxP3（批号315693）、CD3 FITC（批号745694）、γδTCR（批号948037）抗体，中国Abcam公司。
2　实验方法
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2.1　射干麻黄汤水煎剂制备
射干麻黄汤药用：射干9 g，麻黄9 g，细辛6 g，五味子9 g，紫菀9 g，款冬花9 g，姜半夏9 g，大枣10 g，生姜9 g。上述剂量饮片置于烧瓶内加蒸馏水浸泡30 min，再加蒸馏水没过饮片3 cm，电热套煮沸后继续煎煮20 min，用4层无菌纱布过滤药液；加入蒸馏水至没过饮片，电热套煮沸后继续煎煮15 min，用4层无菌纱布过滤药液。合并两次过滤药液，小火浓缩至100 mL，4℃冷藏备用。使用射干麻黄汤期间每7 d制备一次。
2.2　动物分组、干预及采样
2.2.1　过继免疫γδT细胞悬液制备
TCRβ-KO小鼠以2%戊巴比妥钠麻醉后剥离脾脏，置200目滤网上，使用注射器针芯轻柔研磨、过滤，室温1500 r/min、离心半径为10 cm离心10 min后弃上清，裂解红细胞，PBS重复洗涤细胞，得到脾脏单细胞悬液。采用TCRγ/δ＋T细胞分离试剂盒分选出小鼠脾脏γδT细胞，调整细胞浓度在2×105～1×106/mL，备用。
2.2.2　动物造模及样本采集
健康小鼠按体质量区组后使用Excel软件随机分为正常对照组（Ctrl组）、气道高反应模型组（AHR组）、过继免疫γδT细胞组（AIT组）、射干麻黄汤干预组（SGMHD组）、射干麻黄汤＋过继免疫组（SMA组）5组，每组8只。除Ctrl组外，其余小鼠均在第1、7、14天给予0.2 mL致敏溶液（OVA 50μg、氢氧化铝2 mg，0.05 mg/mL）腹腔注射致敏，在第21～27天每天以1% OVA 20μL滴鼻激发，Ctrl组小鼠给予等量1×PBS代替。SGMHD组每日激发后1 h给予射干麻黄汤水煎剂灌胃（0.103 mg/g），AIT组每日激发后1 h经尾静脉注射γδT细胞悬液0.5 mL，SMA组每日激发后1 h给予射干麻黄汤水煎剂灌胃及尾静脉注射γδT细胞悬液。末次干预24 h后给予所有小鼠麻醉（2%戊巴比妥钠0.1 mL/20 g腹腔注射），腹主动脉采血0.5 mL，保存于肝素抗凝管，12 h内行流式细胞学检测；另取0.5 mL腹主动脉血，室温3000 r/min、离心半径为10 cm离心10 min后，取上清置于-80℃保存。麻醉后12 h内剥离小鼠鼻部皮肤，分离双侧鼻腔，分离双侧鼻甲及鼻中隔黏膜，制备4μm石蜡切片。沿胸腹正中线剪开腹腔和胸骨柄，取左侧完整肺组织并置于4%多聚甲醛溶液内固定，用于病理检测。
2.3　观察指标及方法
2.3.1　鼻黏膜及肺组织病理学检测
采用免疫组化法检测鼻黏膜及肺组织中细胞因子IFN-γ、IL-4、视黄酸受体相关孤儿受体γt（retinoic acid receptor-related orphan receptorγt，RORγt）、叉头状转录因子P3（forkhead box protein P3，FoxP3）及γδT细胞受体（γδT cellreceptor，γδTCR）的表达。鼻黏膜及肺组织经4μm石蜡切片，60℃烘烤2 h，二甲苯脱蜡10 min×2次、无水乙醇3 min×2次、95%乙醇2 min×2次、80%乙醇2 min、70%乙醇2 min，高温高压抗原修复，滴加血清封闭20 min后加入相应一抗于4℃孵育过夜。滴加二抗、37℃孵育1 h，二氨基联苯胺显色1 min，自来水冲洗终止反应，苏木素复染1 min，无水乙醇脱水、二甲苯透明、中性树胶封片，Image J2软件分析单位面积阳性细胞表达的平均光密度值。免疫组化染色黄色或棕黄色为阳性，细胞核染为蓝色。
2.3.2　外周血T淋巴细胞亚群检测
采用流式细胞术检测外周血T淋巴细胞亚群水平。抗凝血加入2 mL PBS及淋巴细胞分离液，3000 r/min（离心半径为10 cm）离心10 min并吸取淋巴细胞，重悬于含10%血清RPMI-1640的培养基中，台盼蓝染色确保细胞存活率大于95%。调整细胞浓度至2×105个/mL，分装于24孔培养板，加入10μL佛波酯（12.5μg/mL）及莫能霉素（0.35 mg/mL）混合物后置于37℃、5%CO2培养箱中培养4 h，间隔1 h混匀。流式管收集细胞，PBS重悬后加入5μL鼠单抗CD4 PerCP-Cy5.5及CD8 PE-Cy7抗体，4℃避光孵育30 min，细胞固定、破膜后分别加入5μL IL-4 APC、IFN-γFITC、IL-17 PE、FoxP3 PE-Cy5抗体，再次4℃避光孵育30 min，PBS洗涤并重悬细胞后上机检测。对于γδT细胞检测，在佛波酯及莫能霉素刺激后分别加入鼠单抗CD3 FITC及γδTCR抗体染色。使用FlowJo 10.8软件分析Th1、Th2、Th17、Treg及γδT细胞百分率。
2.3.3　外周血细胞因子检测
外周血上清液放至室温后6000 r/min、离心半径为10 cm离心15 min去除碎片，使用ELISA试剂盒将样品稀释10倍，在已进行酶标包被的96孔板中加入不同浓度标准品及稀释后的样品，检测IL-2、IL-4、IL-5、IL-10、IL-17、IFN-γ及TGF-β在450 nm波长下各孔吸光光度值，并计算样品浓度。操作步骤按照试剂盒说明书进行。
2.3.4　统计学方法
采用GraphPad Prism 9.0进行数据录入及统计，计量资料符合正态分布，使用one-way ANOVA检验，组间多重比较使用Tukey法。P<0.05为差异具有统计学意义。
3　结果
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3.1　鼻黏膜免疫组化染色结果
小鼠鼻黏膜免疫组化结果显示，各组小鼠鼻黏膜IFN-γ、IL-4、FoxP3、RORγt、γδTCR表达差异均存在极显著统计学意义（P<0.001）。其中，AHR组IFN-γ、IL-4、FoxP3、RORγt、γδTCR表达均高于正常对照组（P<0.01或0.001）；与AHR组相比，AIT组、SGMHD组及SMA组鼻黏膜IFN-γ、IL-4、RORγt、γδTCR表达较低（P<0.05、0.01或0.001），AIT组FoxP3表达较低（P<0.05）。见表1。
3.2　肺组织免疫组化染色结果
小鼠肺组织免疫组化结果显示，各组小鼠肺组织IFN-γ、IL-4、FoxP3、RORγt、γδTCR表达差异均存在极显著统计学意义（P<0.001）。其中，AHR组IFN-γ、IL-4、FoxP3、RORγt、γδTCR表达均高于正常对照组（P<0.001）；与AHR组相比，AIT组、SGMHD组及SMA组肺组织IL-4、γδTCR表达较低（P<0.001），AIT组IFN-γ表达较低（P<0.05），SGMHD组、SMA组FoxP3表达较低（P<0.001）。见表2。
3.3　外周血T淋巴细胞亚群检测结果
AHR组小鼠造模成功后外周血Th2、Th17、γδT细胞比例显著高于正常对照组小鼠（P<0.001）；与AHR组相比，AIT组、SGMHD组及SMA组外周血Th1细胞比例较高（P<0.01或0.001），Th17、γδT细胞比例较低（P<0.01或0.001）。AIT组Treg细胞比例低于AHR组（P<0.05）。各组间Th2细胞比例无明显统计学差异。见表3。
3.4　血清细胞因子水平检测结果
与正常对照组相比较，AHR组小鼠外周血IFN-γ、IL-2、IL-4、IL-5及IL-17浓度均较高（P<0.01或0.001），IL-10、TGF-β则显著低于正常对照组（P<0.001）。与AHR组相比，AIT组IFN-γ、IL-2、IL-4及IL-17浓度较高（P<0.05，0.01或0.001），TGF-β浓度较低（P<0.001）。SGMHD组及SMA组IFN-γ、IL-2、IL-4、IL-5及IL-17浓度均显著低于AHR组，IL-10、TGF-β浓度则较高（P<0.001）。见表4。
4　小结与讨论
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本研究通过观察射干麻黄汤对AHR模型小鼠的干预效果，分析射干麻黄汤调控上下气道、外周血淋巴细胞亚群和血清细胞因子水平的作用。实验结果显示，射干麻黄汤可调节AHR模型小鼠的鼻黏膜、肺组织和外周血Th1、Th2、Th17、Treg和γδT细胞亚群比例，纠正细胞比例失衡，并调整多种外周血细胞因子浓度，改善小鼠炎性状态。
目前普遍认为，气道炎症是AHR的首要机制[12]，主要病理表现为气道内嗜酸性粒细胞（eosinophil，EOS）、T淋巴细胞等多种炎症细胞浸润和聚集[13]。2型免疫应答是气道炎症尤其是过敏性气道疾病的发生途径，由Th2及2型固有淋巴细胞产生的IL-4、IL-5等细胞因子诱导EOS和肥大细胞（mast cell，MC）活化，并促使B细胞产生免疫球蛋白E（immunoglobulin E，IgE）。除此之外，Th17细胞及其分泌的IL-17在气道炎症中同样发挥了重要作用，包括诱导中性粒细胞（neutrophil，NEU）和EOS聚集、刺激黏蛋白基因表达从而形成气道黏液高分泌、促进气道周围胶原沉积[14]。而Treg细胞可以通过IL-10、TGF-β依赖方式参与抗炎过程并降低AHR[15]。γδT细胞是T细胞的一种特殊亚群，呈现出气道黏膜优势性增殖[16]，在免疫应答早期γδT细胞作为固有免疫和适应性免疫的桥梁诱导树突状细胞成熟，同时分泌Th1、Th2、Th17相关细胞因子，促使2型炎症反应进展[17]，对AHR的发生具有重要推动作用。
射干麻黄汤出自《金匮要略》，是治疗寒饮咳喘的重要方剂。方中射干、紫菀、款冬、半夏苦泄降气，麻黄、细辛、生姜辛散发表，五味子酸收敛肺，半夏、细辛消痰化饮，大枣调和、安中，诸药配伍，使散中有收，祛邪而不伤正，共奏宣肺祛痰、下气止咳之效。临床上射干麻黄汤常用于治疗典型哮喘、咳嗽变异型哮喘等伴有气道高反应的慢性气道炎症性疾病[18-19]，研究表明射干麻黄汤在治疗气道炎症方面具有多成分、多靶点、多通路的特点。单味药有效成分研究显示，射干麻黄汤中包含野鸢尾黄素、麻黄碱、伪麻黄碱、射干苷、β-谷甾醇、半夏多糖、细辛脂素、紫菀酮、款冬花总倍半萜等[20-22]，其中麻黄碱可通过IL-2、IL-4等细胞因子促进EOS、MC及NEU凋亡，抑制炎症介质释放，减轻气道组织肿胀[23]；β-谷甾醇及半夏多糖可抑制Th2细胞激活并下调其释放的炎症因子，具有显著的抗炎作用[21-22]；细辛脂素可减轻气道嗜酸性粒细胞浸润，降低IgE水平，从而减轻气道高反应性[24-25]。本研究发现射干麻黄汤显著降低AHR模型小鼠鼻黏膜和肺组织IL-4、γδTCR表达水平，提高外周血Th1细胞比例、降低Th17及γδT细胞比例，降低外周血IFN-γ、IL-2、IL-4、IL-5及IL-17水平，并提高IL-10、TGF-β水平。
值得注意的是，本研究显示无论γδT细胞过继免疫、射干麻黄汤治疗还是二者联合干预，与AHR组小鼠相比，仅有鼻黏膜和肺组织中IL-4表达显著降低，各干预组外周血中Th2细胞比例均未出现明显减少，提示射干麻黄汤对AHR的2型免疫反应调节作用可能集中在气道局部。过敏性鼻炎和哮喘是以AHR为主的两种气道慢性炎症，根据“同一气道、同一疾病”概念[26]，近年来提出了过敏性鼻炎哮喘综合征的诊断名称，指同时发生的临床或亚临床上、下呼吸道过敏性症状[27]。《素问·阴阳应象大论篇》曰：“肺主鼻，在窍为鼻”；《诸病源候论》云：“若肺脏调和，则鼻气通利，而知臭香”。鼻在上在外，为肺气出入的通道，肺藏于里，鼻与肺上下相应，相关一体，相互影响。本研究结果提示射干麻黄汤对AHR模型小鼠鼻黏膜和肺组织T淋巴细胞亚群具有同向调节作用，可抑制γδT细胞、Th2细胞介导的免疫应答，为其“鼻肺同治”的现代科学内涵提供了理论基础。
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2025年第43卷第11期
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doi: 10.13193/j.issn.1673-7717.2025.11.015
  • 首发时间：2026-10-30
  • 出版时间：2025-11-10
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国家自然科学基金项目(82174448; 82374529)
北京市自然科学基金面上项目(7222269)
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    首都医科大学附属北京中医医院，北京 100010

通讯作者:

王俊阁（1967-），男，河北沧州人，主任医师、教授，博士研究生导师，博士，研究方向：经方治疗气道变态反应性疾病。E-mail：。
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2种不同金属材料的力学参数
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Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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