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Article(id=1322926843229250487, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322926740955341011, articleNumber=null, orderNo=null, doi=10.13193/j.issn.1673-7717.2025.10.045, 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=1793339938218, onlineDateStr=2026-10-30, pubDate=1760025600000, pubDateStr=2025-10-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793339938218, onlineIssueDateStr=2026-10-30, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793339938218, creator=13701087609, updateTime=1793339938218, updator=13701087609, issue=Issue{id=1322926740955341011, tenantId=1146029695717560320, journalId=1255847919539208197, year='2025', volume='43', issue='10', pageStart='1', pageEnd='258', issueExtLink='null', onlineDate='null', pubDate='1760025600000', pubDateStr='2025-10-10', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1793339913834, creator='13701087609', updateTime=1793340753123, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1322930261259846420, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322926740955341011, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1322930261259846421, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322926740955341011, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=237, endPage=242, ext={EN=ArticleExt(id=1322926843464131512, articleId=1322926843229250487, tenantId=1146029695717560320, journalId=1255847919539208197, language=EN, title=Basic Study on Regulation of Th2 Immune Response in Rats with Allergic Rhinitis by Ebushicao（Centipeda minima）Extract via Mediating Toll-like Receptor-Nuclear Factor Kappa B Signaling Pathway, columnId=1256183365644968140, journalTitle=Chinese Archives of Traditional Chinese Medicine, columnName=Platform of Provincial Project, runingTitle=null, highlight=null, articleAbstract=
Objective

To study the regulation of T helper cell 2（TH2）immune response in rats with allergic rhinitis（AR）by Ebushicao（Centipeda minima）extract via mediating Toll-like receptor-nuclear factor kappa B（TLR/NF-κB）signaling pathway.

Methods

ARmodels were induced with ovalbumin（OVA），and the rats were divided into model group，positive control group and Ebushicao（Centipeda minima）extract treatment groups（low，middle，high concentrations）.The rats in the positive control group were given loratadine（2 mg/kg）by gavage.The rats in the Ebushicao（Centipeda minima）extract treatment groups were given Ebushicao（Centipeda minima）extracts with the corresponding doses（2 mg/kg，4 mg/kg，8 mg/kg）by gavage，while those in the blank control and model groups were given an equal volume of saline by gavage.The treatment in all groups lasted for 10 days.Weights of nasal secretions were collected from the rats in each group and the scores for nasal symptoms were calculated.HE staining was used to observe the morphological changes of nasal mucosa，and the counts of eosinophils and neutrophils in nasal mucosa were calculated.The levels of interleukin-4（IL-4），interleukin-13（IL-13）and immunoglobulin E（IgE）in peripheral blood of rats in each group were detected by ELISA.The mRNA and protein expression levels of TLR-4 and NF-κB were detected by RT-PCR and Western-blot.In Ebushicao（Centipeda minima）extract treatment groups，the rats received natural TLR4 agonist lipopolysaccharide（LPS）and were observed for morphology of the nasal mucosa and nasal symptoms.

Results

Compared with the blank control group，the rats in the model group had a significant increase in behavioral score of nasal symptoms，the levels of serum IL-4，IL-13 and IgE，the levels of eosinophil and neutrophi in nasal mucosal tissues，and the relative mRNA and protein expressions of TLR4 and NF-κB in nasal mucosal tissues，with statistical difference（P<0.05），while the above increases were notably diminished to varying degrees in the positive control group and Ebushicao（Centipeda minima）extract treatment groups（P<0.05）.Within Ebushicao（Centipeda minima）extract treatment groups，a gradual decline was observed in the above indexes as the concentration increased，with statistical difference in paired comparison（all P<0.05）.HE staining showed that the nasal mucosa of the control rats was normal pseudostratified ciliated columnar epithelium with intact mucosal epithelial structure and there was no obvious inflammatory cell infiltration.The epithelium of the nasal mucosa of the rats in the model group was heavily detached，and a large number of eosinophils and mast cells were observed in the lamina propria of the mucosa.Various degrees of pathological changes were also found in positive control group and Ebushicao（Centipeda minima）extract treatment groups，while the changes were remarkably attenuated compared with those in the model group.The weakening of pathological changes showed a concentration dependent manner within Ebushicao（Centipeda minima）extract treatment groups.After TLR4 agonist treatment，the improved AR symptoms and pathological changes in Ebushicao（Centipeda minima）extract treatment groups were reversed.

Conclusion

The mechanism of Ebushicao（Centipeda minima）extract in the treatment of AR may be related to the inhibition of TLR/NF-κB signaling pathway and alleviation of TH2 immune response.

, authors=Jun LIU, Chengyi SONG, Jianle XU, Zhuoping LIANG, authorsList=Jun LIU, Chengyi SONG, Jianle XU, Zhuoping LIANG, authorCompany=null, correspAuthors=null, 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=1322926848124003286, articleId=1322926843229250487, tenantId=1146029695717560320, journalId=1255847919539208197, language=CN, title=鹅不食草提取物介导Toll样受体通路调节变应性鼻炎大鼠Th2型免疫反应的基础研究, columnId=1256183369033965799, journalTitle=中华中医药学刊, columnName=省级项目平台, runingTitle=null, highlight=null, articleAbstract=
目的

研究鹅不食草提取物介导Toll样受体核因子通路（Toll-like receptor/nuclearfactor-κB，TLR/NF-κB）调节变应性鼻炎（allergic rhinitis，AR）大鼠Th2型免疫反应。

方法

卵清白蛋白（ovalbumin，OVA）建立AR大鼠模型，建模成功后将其分为模型组、阳性对照组及鹅不食草提取物不同浓度组（低浓度、中浓度、高浓度），其中阳性对照组给予氯雷他定（2 mg/kg）灌胃，鹅不食草提取物组大鼠分别给予2、4、8 mg/kmg鹅不食草提取物灌胃，对照组及模型组大鼠给予等体积生理盐水灌胃，连续灌胃10 d。收集各组大鼠鼻腔分泌物重量，统计其鼻部症状评分，HE染色观察各组大鼠鼻黏膜组织形态学改变、计算鼻黏膜内嗜酸性粒细胞及中性粒细胞计数，酶联免疫吸附法检测各组大鼠外周血白细胞介素-4（interleukin-4，IL-4）、白细胞介素-13（interleukin，IL-13）、免疫球蛋白E（immunoglobulin E，IgE）水平。RTPCR法及Western-Blot检测鼻腔黏膜组织TLR-4及NF-κB mRNA及蛋白表达水平。在不同浓度鹅不食草提取物组大鼠中加用TLR4激动剂LPS，观察大鼠鼻黏膜形态学、鼻部症状行为评分等。

结果

与对照组相比，AR模型组大鼠鼻腔症状行为学得分，血清IL-4、IL-13及IgE水平，鼻腔黏膜组织内嗜酸性粒细胞、中性粒细胞水平，以及鼻腔黏膜组织内TLR4及NF-κB相对mRNA及蛋白表达量均明显升高，差异具有统计学意义（P<0.05）。与模型组相比，鹅不食草提取物不同浓度组及阳性对照组大鼠以上检测指标水平均有不同程度的下降，差异均具有统计学意义（P<0.05）。且随着鹅不食草提取物浓度的上升，大鼠以上指标水平均依次下降，组间两两比较，差异均存在统计学意义（P<0.05）。HE染色可见对照组大鼠鼻腔黏膜组织为正常假复层纤毛柱状上皮，黏膜上皮结构完整，排列整齐，未见明显炎性细胞浸润，而模型组大鼠鼻腔黏膜组织上皮大量脱落，黏膜固层可见大量嗜酸性粒细胞及肥大细胞浸润，鹅不食草提取物不同浓度组及阳性对照组大鼠鼻腔黏膜组织上皮有不同程度的AR病理改变表现，但其严重程度均低于模型组，且随着鹅不食草提取物浓度的上升，AR大鼠鼻腔黏膜病变程度越低。经TLR4激动剂处理后，鹅不食草提取物不同浓度组AR大鼠已好转的AR相关症状及病理改变均向更严重的方向逆转。

结论

鹅不食草提取物可通过抑制TLR/NF-κB通路，减轻Th2型免疫反应强度，治疗变应性鼻炎。

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刘均（1985-），男，四川泸县人，主治医师，硕士，研究方向：咽喉科学。

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刘均（1985-），男，四川泸县人，主治医师，硕士，研究方向：咽喉科学。

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刘均（1985-），男，四川泸县人，主治医师，硕士，研究方向：咽喉科学。

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注：A、B、C分别为鹅不食草提取物低、中、高浓度组。*与对照组比较，P<0.05；＃与模型组比较，P<0.05；a与鹅不食草提取物低浓度组比较，P<0.05；b与鹅不食草提取物中浓度组比较，P<0.05。

, figureFileSmall=IasRG6NeQv63hKwNZHmcZQ==, figureFileBig=AV5LnGJ49P0P4NYjmu+5RA==, tableContent=null), ArticleFig(id=1322926852389610490, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=EN, label=null, caption=null, figureFileSmall=jzWX+3Q6Hj8hed3L/U9n2Q==, figureFileBig=HKeNep3WbVvOpcrGh4pdiQ==, tableContent=null), ArticleFig(id=1322926852460913659, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=CN, label=图3, caption=各组大鼠NF-κB相对mRNA表达量

注：A、B、C分别为鹅不食草提取物低、中、高浓度组。*与对照组比较，P<0.05；＃与模型组比较，P<0.05；a与鹅不食草提取物低浓度组比较，P<0.05；b与鹅不食草提取物中浓度组比较，P<0.05。

, figureFileSmall=jzWX+3Q6Hj8hed3L/U9n2Q==, figureFileBig=HKeNep3WbVvOpcrGh4pdiQ==, tableContent=null), ArticleFig(id=1322926852540605436, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=EN, label=null, caption=null, figureFileSmall=PPp7/X/7WQGT5D/xX4chEA==, figureFileBig=xa6Var453KNZ/m0WdjcFTw==, tableContent=null), ArticleFig(id=1322926852616102909, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=CN, label=图4, caption=各组大鼠鼻黏膜组织内TLR4及NF-κB蛋白表达量

注：A、B、C分别为鹅不食草提取物低、中、高浓度组。

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注：＃与未使用TLR4激动剂处理比较，P<0.05。

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注：＃与未使用TLR4激动剂处理比较，P<0.05。

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注：A、B、C分别为鹅不食草提取物低、中、高浓度组。

, figureFileSmall=Eea6Wr8sUC6Nfw3lS76FRQ==, figureFileBig=ihBUnJmBKfsS8QDoFO4NEQ==, tableContent=null), ArticleFig(id=1322926853215887363, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别n鼻腔症状行为学得分/（分）
对照组101.65±0.33
模型组109.86±1.47*
鹅不食草提取物低浓度组107.11±1.16#
鹅不食草提取物中浓度组105.89±1.21#a
鹅不食草提取物高浓度组104.61±1.19#ab
阳性对照组104.36±1.23#
), ArticleFig(id=1322926853287190532, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=CN, label=表1, caption=

各组大鼻腔症状行为学得分比较（）

, figureFileSmall=null, figureFileBig=null, tableContent=
组别n鼻腔症状行为学得分/（分）
对照组101.65±0.33
模型组109.86±1.47*
鹅不食草提取物低浓度组107.11±1.16#
鹅不食草提取物中浓度组105.89±1.21#a
鹅不食草提取物高浓度组104.61±1.19#ab
阳性对照组104.36±1.23#
), ArticleFig(id=1322926853358493701, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别nIL-4/（pg/mL）IL-13/（ng/L）IgE/（KU/L）
对照组1031.26±4.6943.58±6.991.21±0.21
模型组1066.84±8.49*83.55±13.25*2.63±0.41*
鹅不食草提取物低浓度组1053.65±6.83#71.15±10.15#2.14±0.37#
鹅不食草提取物中浓度组1042.15±6.61#a60.12±12.74#a1.76±0.28#a
鹅不食草提取物高浓度组1036.51±5.79#ab45.69±8.97#ab1.53±0.24#ab
阳性对照组1035.15±5.34#46.11±9.73#1.41±0.31#
), ArticleFig(id=1322926853438185478, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=CN, label=表2, caption=

各组大鼠血清IL-4、IL-13、IgE水平比较（）

, figureFileSmall=null, figureFileBig=null, tableContent=
组别nIL-4/（pg/mL）IL-13/（ng/L）IgE/（KU/L）
对照组1031.26±4.6943.58±6.991.21±0.21
模型组1066.84±8.49*83.55±13.25*2.63±0.41*
鹅不食草提取物低浓度组1053.65±6.83#71.15±10.15#2.14±0.37#
鹅不食草提取物中浓度组1042.15±6.61#a60.12±12.74#a1.76±0.28#a
鹅不食草提取物高浓度组1036.51±5.79#ab45.69±8.97#ab1.53±0.24#ab
阳性对照组1035.15±5.34#46.11±9.73#1.41±0.31#
), ArticleFig(id=1322926853534654471, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别n 嗜酸性粒细胞计数中性粒细胞计数
对照组100.31±0.041.25±0.23
模型组1015.15±3.15*4.63±1.46*
鹅不食草提取物低浓度组1011.15±2.17#3.65±0.89#
鹅不食草提取物中浓度组106.33±1.13#a2.71±0.53#a
鹅不食草提取物高浓度组104.15±1.04#ab1.65±0.41#ab
阳性对照组104.04±1.22#1.74±0.27#
), ArticleFig(id=1322926853647900680, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=CN, label=表3, caption=

光学显微镜下各组大鼠鼻腔黏膜组织内嗜酸性粒细胞、中性粒细胞计数比较（）

, figureFileSmall=null, figureFileBig=null, tableContent=
组别n 嗜酸性粒细胞计数中性粒细胞计数
对照组100.31±0.041.25±0.23
模型组1015.15±3.15*4.63±1.46*
鹅不食草提取物低浓度组1011.15±2.17#3.65±0.89#
鹅不食草提取物中浓度组106.33±1.13#a2.71±0.53#a
鹅不食草提取物高浓度组104.15±1.04#ab1.65±0.41#ab
阳性对照组104.04±1.22#1.74±0.27#
), ArticleFig(id=1322926853723398153, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别nTLR4NF-κΒ
对照组100.32±0.050.21±0.06
模型组102.15±0.52*1.33±0.38*
A组100.81±0.13#0.71±0.15#
B组100.53±0.10#a0.47±0.11#a
C组100.41±0.07#ab0.38±0.09#ab
阳性对照组100.33±0.04#0.23±0.05#
), ArticleFig(id=1322926853828255754, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=CN, label=表4, caption=

Western-Blot法检测各组大鼠鼻黏膜组织内TLR4及NF-κΒ蛋白表达量（）

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组别nTLR4NF-κΒ
对照组100.32±0.050.21±0.06
模型组102.15±0.52*1.33±0.38*
A组100.81±0.13#0.71±0.15#
B组100.53±0.10#a0.47±0.11#a
C组100.41±0.07#ab0.38±0.09#ab
阳性对照组100.33±0.04#0.23±0.05#
), ArticleFig(id=1322926853907947531, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别nTLR4激动剂处理鼻腔症状行为学得分/（分）嗜酸性粒细胞计数/（109L）中性粒细胞计数/（109L）IL-4/（pg/mL）IL-13/（ng/L）IgE/（KU/L）
鹅不食草提取物低浓度组10否7.11±1.1611.15±2.173.65±0.8953.65±6.8371.15±10.152.14±0.37
是8.79±1.16#13.98±2.74#4.17±1.05#60.48±12.54#79.46±13.69#2.49±0.32#
鹅不食草提取物中浓度组10否5.89±1.216.33±1.132.71±0.5342.15±6.6160.12±12.741.76±0.28
是7.23±1.56#8.96±2.41#3.63±0.84#53.68±10.77#70.55±12.71#2.35±0.31#
鹅不食草提取物高浓度组10否4.61±1.194.15±1.041.65±0.4136.51±5.7945.69±8.971.53±0.24
是6.79±1.43#6.84±2.07#2.84±0.45#45.77±8.97#53.33±10.58#2.09±0.27#
), ArticleFig(id=1322926855577280524, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=CN, label=表5, caption=

TLR4激动剂对鹅不食草提取物处理组AR大鼠的影响（）

, figureFileSmall=null, figureFileBig=null, tableContent=
组别nTLR4激动剂处理鼻腔症状行为学得分/（分）嗜酸性粒细胞计数/（109L）中性粒细胞计数/（109L）IL-4/（pg/mL）IL-13/（ng/L）IgE/（KU/L）
鹅不食草提取物低浓度组10否7.11±1.1611.15±2.173.65±0.8953.65±6.8371.15±10.152.14±0.37
是8.79±1.16#13.98±2.74#4.17±1.05#60.48±12.54#79.46±13.69#2.49±0.32#
鹅不食草提取物中浓度组10否5.89±1.216.33±1.132.71±0.5342.15±6.6160.12±12.741.76±0.28
是7.23±1.56#8.96±2.41#3.63±0.84#53.68±10.77#70.55±12.71#2.35±0.31#
鹅不食草提取物高浓度组10否4.61±1.194.15±1.041.65±0.4136.51±5.7945.69±8.971.53±0.24
是6.79±1.43#6.84±2.07#2.84±0.45#45.77±8.97#53.33±10.58#2.09±0.27#
), ArticleFig(id=1322926855665360909, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别TLR4激动剂（LPS）处理TLR4蛋白表达NF-κΒ蛋白表达
鹅不食草提取物低浓度组否0.71±0.121.61±0.33
是1.16±0.33#2.23±0.41#
鹅不食草提取物中浓度组否0.43±0.080.77±0.15
是0.84±0.15#1.77±0.26#
鹅不食草提取物高浓度组否0.21±0.040.46±0.11
是0.59±0.08#1.38±0.32#
), ArticleFig(id=1322926855732469774, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322926843229250487, language=CN, label=表6, caption=

TLR4激动剂对鹅不食草提取物处理组AR大鼠TLR4、NF-κΒ蛋白表达的影响（）

, figureFileSmall=null, figureFileBig=null, tableContent=
组别TLR4激动剂（LPS）处理TLR4蛋白表达NF-κΒ蛋白表达
鹅不食草提取物低浓度组否0.71±0.121.61±0.33
是1.16±0.33#2.23±0.41#
鹅不食草提取物中浓度组否0.43±0.080.77±0.15
是0.84±0.15#1.77±0.26#
鹅不食草提取物高浓度组否0.21±0.040.46±0.11
是0.59±0.08#1.38±0.32#
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鹅不食草提取物介导Toll样受体通路调节变应性鼻炎大鼠Th2型免疫反应的基础研究
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刘均 , 宋程懿 , 徐建乐 , 梁灼萍
中华中医药学刊 | 省级项目平台 2025,43(10): 237-242
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中华中医药学刊 |省级项目平台 2025 , 43 (10) : 237 -242
鹅不食草提取物介导Toll样受体通路调节变应性鼻炎大鼠Th2型免疫反应的基础研究
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刘均, 宋程懿, 徐建乐, 梁灼萍
作者信息
  • 西南医科大学附属医院，四川 泸州 646000
作者简介:

刘均（1985-），男，四川泸县人，主治医师，硕士，研究方向：咽喉科学。

Basic Study on Regulation of Th2 Immune Response in Rats with Allergic Rhinitis by Ebushicao（Centipeda minima）Extract via Mediating Toll-like Receptor-Nuclear Factor Kappa B Signaling Pathway
Jun LIU, Chengyi SONG, Jianle XU, Zhuoping LIANG
Affiliations
  • The Affiliated Hospital of Southwest Medical University，Luzhou 646000，Sichuan，China
出版时间: 2025-10-10 doi: 10.13193/j.issn.1673-7717.2025.10.045
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摘要
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目的

研究鹅不食草提取物介导Toll样受体核因子通路（Toll-like receptor/nuclearfactor-κB，TLR/NF-κB）调节变应性鼻炎（allergic rhinitis，AR）大鼠Th2型免疫反应。

方法

卵清白蛋白（ovalbumin，OVA）建立AR大鼠模型，建模成功后将其分为模型组、阳性对照组及鹅不食草提取物不同浓度组（低浓度、中浓度、高浓度），其中阳性对照组给予氯雷他定（2 mg/kg）灌胃，鹅不食草提取物组大鼠分别给予2、4、8 mg/kmg鹅不食草提取物灌胃，对照组及模型组大鼠给予等体积生理盐水灌胃，连续灌胃10 d。收集各组大鼠鼻腔分泌物重量，统计其鼻部症状评分，HE染色观察各组大鼠鼻黏膜组织形态学改变、计算鼻黏膜内嗜酸性粒细胞及中性粒细胞计数，酶联免疫吸附法检测各组大鼠外周血白细胞介素-4（interleukin-4，IL-4）、白细胞介素-13（interleukin，IL-13）、免疫球蛋白E（immunoglobulin E，IgE）水平。RTPCR法及Western-Blot检测鼻腔黏膜组织TLR-4及NF-κB mRNA及蛋白表达水平。在不同浓度鹅不食草提取物组大鼠中加用TLR4激动剂LPS，观察大鼠鼻黏膜形态学、鼻部症状行为评分等。

结果

与对照组相比，AR模型组大鼠鼻腔症状行为学得分，血清IL-4、IL-13及IgE水平，鼻腔黏膜组织内嗜酸性粒细胞、中性粒细胞水平，以及鼻腔黏膜组织内TLR4及NF-κB相对mRNA及蛋白表达量均明显升高，差异具有统计学意义（P<0.05）。与模型组相比，鹅不食草提取物不同浓度组及阳性对照组大鼠以上检测指标水平均有不同程度的下降，差异均具有统计学意义（P<0.05）。且随着鹅不食草提取物浓度的上升，大鼠以上指标水平均依次下降，组间两两比较，差异均存在统计学意义（P<0.05）。HE染色可见对照组大鼠鼻腔黏膜组织为正常假复层纤毛柱状上皮，黏膜上皮结构完整，排列整齐，未见明显炎性细胞浸润，而模型组大鼠鼻腔黏膜组织上皮大量脱落，黏膜固层可见大量嗜酸性粒细胞及肥大细胞浸润，鹅不食草提取物不同浓度组及阳性对照组大鼠鼻腔黏膜组织上皮有不同程度的AR病理改变表现，但其严重程度均低于模型组，且随着鹅不食草提取物浓度的上升，AR大鼠鼻腔黏膜病变程度越低。经TLR4激动剂处理后，鹅不食草提取物不同浓度组AR大鼠已好转的AR相关症状及病理改变均向更严重的方向逆转。

结论

鹅不食草提取物可通过抑制TLR/NF-κB通路，减轻Th2型免疫反应强度，治疗变应性鼻炎。

鹅不食草提取物  /  Toll样受体核因子通路  /  变应性鼻炎大鼠  /  Th2型免疫反应
Objective

To study the regulation of T helper cell 2（TH2）immune response in rats with allergic rhinitis（AR）by Ebushicao（Centipeda minima）extract via mediating Toll-like receptor-nuclear factor kappa B（TLR/NF-κB）signaling pathway.

Methods

ARmodels were induced with ovalbumin（OVA），and the rats were divided into model group，positive control group and Ebushicao（Centipeda minima）extract treatment groups（low，middle，high concentrations）.The rats in the positive control group were given loratadine（2 mg/kg）by gavage.The rats in the Ebushicao（Centipeda minima）extract treatment groups were given Ebushicao（Centipeda minima）extracts with the corresponding doses（2 mg/kg，4 mg/kg，8 mg/kg）by gavage，while those in the blank control and model groups were given an equal volume of saline by gavage.The treatment in all groups lasted for 10 days.Weights of nasal secretions were collected from the rats in each group and the scores for nasal symptoms were calculated.HE staining was used to observe the morphological changes of nasal mucosa，and the counts of eosinophils and neutrophils in nasal mucosa were calculated.The levels of interleukin-4（IL-4），interleukin-13（IL-13）and immunoglobulin E（IgE）in peripheral blood of rats in each group were detected by ELISA.The mRNA and protein expression levels of TLR-4 and NF-κB were detected by RT-PCR and Western-blot.In Ebushicao（Centipeda minima）extract treatment groups，the rats received natural TLR4 agonist lipopolysaccharide（LPS）and were observed for morphology of the nasal mucosa and nasal symptoms.

Results

Compared with the blank control group，the rats in the model group had a significant increase in behavioral score of nasal symptoms，the levels of serum IL-4，IL-13 and IgE，the levels of eosinophil and neutrophi in nasal mucosal tissues，and the relative mRNA and protein expressions of TLR4 and NF-κB in nasal mucosal tissues，with statistical difference（P<0.05），while the above increases were notably diminished to varying degrees in the positive control group and Ebushicao（Centipeda minima）extract treatment groups（P<0.05）.Within Ebushicao（Centipeda minima）extract treatment groups，a gradual decline was observed in the above indexes as the concentration increased，with statistical difference in paired comparison（all P<0.05）.HE staining showed that the nasal mucosa of the control rats was normal pseudostratified ciliated columnar epithelium with intact mucosal epithelial structure and there was no obvious inflammatory cell infiltration.The epithelium of the nasal mucosa of the rats in the model group was heavily detached，and a large number of eosinophils and mast cells were observed in the lamina propria of the mucosa.Various degrees of pathological changes were also found in positive control group and Ebushicao（Centipeda minima）extract treatment groups，while the changes were remarkably attenuated compared with those in the model group.The weakening of pathological changes showed a concentration dependent manner within Ebushicao（Centipeda minima）extract treatment groups.After TLR4 agonist treatment，the improved AR symptoms and pathological changes in Ebushicao（Centipeda minima）extract treatment groups were reversed.

Conclusion

The mechanism of Ebushicao（Centipeda minima）extract in the treatment of AR may be related to the inhibition of TLR/NF-κB signaling pathway and alleviation of TH2 immune response.

Ebushicao（Centipeda minima）extract  /  Toll-like receptor-nuclear factor kappa B signaling pathway  /  rats with allergic rhinitis  /  Th2 immune response
刘均, 宋程懿, 徐建乐, 梁灼萍. 鹅不食草提取物介导Toll样受体通路调节变应性鼻炎大鼠Th2型免疫反应的基础研究. 中华中医药学刊, 2025 , 43 (10) : 237 -242 . DOI: 10.13193/j.issn.1673-7717.2025.10.045
Jun LIU, Chengyi SONG, Jianle XU, Zhuoping LIANG. Basic Study on Regulation of Th2 Immune Response in Rats with Allergic Rhinitis by Ebushicao（Centipeda minima）Extract via Mediating Toll-like Receptor-Nuclear Factor Kappa B Signaling Pathway[J]. Chinese Archives of Traditional Chinese Medicine, 2025 , 43 (10) : 237 -242 . DOI: 10.13193/j.issn.1673-7717.2025.10.045
正文
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变应性鼻炎（allergic rhinitis，AR）是最常见的鼻部疾病，随着我国近年来工业事业的不断发展，AR发病率呈逐年上升趋势，AR与许多其他呼吸道疾病密切相关，对患者生活及学习均存在一定的不良影响[1]。AR以免疫球蛋白E（immunoglobulin E，IgE）介导的嗜酸性粒细胞反应为主，具有明显的Th2型免疫反应特征[2-3]。Toll样受体（Toll-like receptors，TLRs）家族参与了病原体识别与结合，可介导体液免疫，诱导炎症反应，核因子（nuclear factor-κB，NF-κB）位于TLR下游信号通路枢纽位置，TLR/NF-κB信号途径参与了细胞免疫应答、炎症反应，是调节Th2型免疫反应的重要信号通路[4-5]。
鹅不食草是我国传统中药材，具有辛温解表，通窍止咳之效，常用于治疗鼻炎[6]。鹅不食草内可提取出蒲公英甾醇醋酸酯、单萜、多萜、百里酚衍生物等多种成分，有实验表明，鹅不食草提取物可有效阻断组胺释放，在被动皮肤过敏实验中具有较强抗敏活性[7-8]。但关于鹅不食草提取物在治疗AR中的效果及其作用原理依旧不明确，为研究鹅不食草提取物是否可通过Toll样受体核因子通路调节Th2型免疫反应，减轻AR相关症状，我们开展如下研究。
1　材料与方法
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1.1　材料
1.1.1　实验动物
健康Wistar大鼠60只，雌雄不限，体质量250～300 g，购自成都菱祐生物科技有限公司。实验前先适应性喂养1周。
1.1.2　实验试剂及仪器
卵清白蛋白（ovalbumin，OVA）购自美国Sigma-Aldrich公司，鹅不食草购自华东医药股份有限公司，产地浙江，经相关机构鉴定符合《中华人民共和国药典》中要求，为合格品。氯雷他定购自浙江东亚药业股份有限公司。TLR4激动剂LPS购自美国AnaSpec公司。
苏木精-伊红试剂盒购自北京索莱宝生物有限公司，大鼠IL-4、IL-13、IgE ELISA试剂盒购自上海依科赛公司。TRIzol试剂盒购自美国Invitrogen公司，PrimeScript RT试剂盒、SYBR Prime Script RT-PCT试剂盒均购自Takara公司，TLR-4抗体及NF-κB抗体购自英国Abcam公司。实时定量荧光聚合酶链式反应仪（Real-time PCR）购自美国ABI公司，显微镜购自日本Olympus公司。
1.1.3　鹅不食草提取物制备[9]
称取鹅不食草1 kg，适当粉碎，使用80%乙醇浸泡48 h，以药材10倍量的80%乙醇渗漉提取，提取液减压浓缩，回收乙醇后浓缩液，加入乙酸乙酯，醇提后药渣以水煎提取，得到相应提取物，浓缩至浸膏，放置于减压干燥箱内，60℃干燥至无溶剂残留。临时取定量提取物，将其溶解于二甲基亚砜中，全部溶解后，加入RPMⅠ-1640培养液，分别将提取物浓度配置为2、4、8 mg/mL。
1.2　方法
1.2.1　建模方法[10]
将60只Wistar大鼠随机均分为6组，即对照组、模型组、鹅不食草不同浓度提取物组（低浓度、中浓度、高浓度），阳性对照组，每组均10只。适应性喂养1周，除对照组大鼠外，其余大鼠分别于第1、3、5、7、9、11、13天，使用0.3 mg AVO+Al（OH）3 30 mg混合于1 mL生理盐水，腹腔注射致敏，第15天开始鼻腔激发，向试验大鼠两侧鼻腔内各滴加50μL OVA（100μg/100μL）强化致敏，连续滴加7 d。对照组大鼠以同体积生理盐水行腹腔注射与滴鼻。
末次造模结束后，观察各组大鼠30 min内喷嚏、抓鼻及流涕等行为情况，根据赵秀杰等[7]制定的量表进行定量，其中喷嚏1～3次、4～10次、≥11次分别计1、2、3分，抓鼻1～5次、6～15次、≥16次分别计1、2、3分，流涕至鼻前孔、流涕出鼻前孔、流涕满面分别得1、2、3分。各项得分相加为总得分，总得分>5为AR建模成功。
1.2.2　各组大鼠处理方法
阳性对照组给予氯雷他定（2 mg/kg）灌胃，鹅不食草提取物组大鼠分别给予2、4、8 mg/kg鹅不食草提取物灌胃，对照组及模型组大鼠给予等体积生理盐水灌胃，连续灌胃10 d。
1.2.3　TLR4激动剂对鹅不食草提取物处理组AR大鼠的影响
连续给药灌胃处理后，分别在不同浓度鹅不食草提取物组中取5只大鼠，对其腹腔注射TLR4激动剂LPS（0.12 mg/g），观察鹅不食草提取物各浓度组大鼠鼻腔症状行为学、鼻黏膜组织等改变。
1.2.4　各组大鼠鼻腔症状行为学观察[11]
给药干预后，统计各组大鼠30 min内喷嚏、抓鼻及流涕等行为得分。
1.2.5　血清IL-4、IL-13、IgE水平检测
处死大鼠前，抽取其尾部静脉血2 mL，3000 r/min离心15 min，分离上层血清，ELISA法检测血清L-4、IL-13、IgE水平，严格按照试剂盒相关标准进行操作。
1.2.6　鼻黏膜组织HE染色光学显微镜观察[12]
腹腔注射10%水合氯醛（100μL/g）麻醉，开胸暴露心脏，向左心室灌注生理盐水，鼻背正中缝裂开鼻腔，剥离下鼻黏膜组织，10%多聚甲醛固定，连续石蜡切片，行HE染色，光学显微镜下观察大鼠鼻黏膜标本组织结构改变，在40倍物镜下行嗜酸性粒细胞计数、中性粒细胞计数。
1.2.7　RT-PCR检测鼻黏膜组织内TLR4及NF-κB相对mRNA表达[13]
取各组大鼠鼻腔黏膜组织，加入300μL TRNzol裂解组织，逆转录为mRNA，逆转录反应总体系是20μL，但各项总合不够20μL：纯水5μL，MgCl2 4μL，10×Buffer 2μL，dNTP 2μL，逆转录酶0.5μL，核苷酸抑制酶0.5μL，加入RevertAid TM M-MulV Reverse Transciptase（200 U/μL）1μL，42℃发育60 min，70℃，10 min后终止反应，cDNA存放在-80℃冰箱待用。RT-PCR反应总体系20μL：PCR-mix 10μL，纯水8μL，上下引物各0.5μL，模板CDNA 1μL。引物序列由上海博尚Biosune生物有限公司合成，TLR4上游引物：5’-GGGACTCTGATCATGGCATT-3’，下游引物：5’-GTCTCCACAGCCACCAGATT-3’，扩增长度380 bp。NF-κB上游引物：5’-GCTCGGCTGAATGAATCTACCA-3’，下游引物：5’-GTCTCCACGTATTTCCGCAACT-3’，扩增长度168 bp，β-Actin上游引物：5’-GGCATCCTGACCCTGAAGTA-3’，下游引物：5’-AGGAAGGAAGGCTGGAAGAG-3’，扩增长度614 bp。反应条件：50℃，2 min，95℃，2 min，95℃，30 s，60℃，30 s，35个循环。以β-Actin为内参，计算目的基因相对mRNA表达量。
1.2.8　Western-Blot法检测鼻黏膜组织内TLR4及NF-κB蛋白表达量
取各组大鼠鼻腔黏膜组织，加入蛋白裂解液，按照试剂盒相关标准提取鼻腔黏膜组织总蛋白，取等质量总蛋白电泳，转移至PVDF膜，5%脱脂奶粉室温封闭2 h，分别加入兔源抗TLR4（1∶200）、NF-κB（1∶400）、β-Actin抗体（1∶4000），4℃孵育过夜，加入羊抗兔二抗，室温孵育1 h，增强化学发光法显色，凝胶成像系统拍照，Quantity One软件分析各组蛋白灰度值，目的蛋白相对表达量＝目的蛋白灰度值/内参蛋白灰度值。
1.3　统计学方法
采用SPSS 19.0统计软件处理数据。计量资料以表示，多组间比较使用单方差分析，检验有意义者，两两比较采用LSD-t检验，以P<0.05为差异有统计学意义。
2　结果
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2.1　各组大鼠鼻腔症状行为学得分比较
与对照组相比，AR模型组大鼠鼻腔症状行为学得分明显升高，差异具有统计学意义（P<0.05）。与模型组相比，鹅不食草提取物不同浓度组及阳性对照组大鼠鼻腔症状行为得分均有不同程度的下降，差异均具有统计学意义（P<0.05）。且随着鹅不食草提取物浓度的上升，大鼠鼻腔症状行为学得分依次下降，组间两两比较，差异均有统计学意义（P<0.05）。见表1。
2.2　各组大鼠血清IL-4、IL-13、IgE水平比较
与对照组相比，AR模型组大鼠血清IL-4、IL-13及IgE水平明显升高，差异具有统计学意义（P<0.05）。与模型组相比，鹅不食草提取物不同浓度组及阳性对照组大鼠血清IL-4、IL-13及IgE水平均有不同程度的下降，差异均具有统计学意义（P<0.05）。且随着鹅不食草提取物浓度的上升，大鼠血清IL-4、IL-13及IgE水平依次下降，组间两两比较，差异均存在统计学意义（P<0.05）。见表2。
2.3　各组大鼠鼻腔黏膜组织HE染色结果比较
对照组大鼠鼻腔黏膜组织为正常假复层纤毛柱状上皮，黏膜上皮结构完整，排列整齐，未见明显炎性细胞浸润。见插页ⅩⅩⅩⅣ图1A。
模型组大鼠鼻腔黏膜组织上皮有脱落现象，不完整，黏膜固层可见大量嗜酸性粒细胞及肥大细胞浸润。见插页ⅩⅩⅩⅣ图1B。
鹅不食草提取物不同浓度组大鼠及阳性对照组大鼠鼻腔黏膜组织上皮有不同程度脱落现象，黏膜固层可见嗜酸性粒细胞及肥大细胞浸润，但其浸润严重程度均低于模型组，且随着鹅不食草提取物浓度的上升，AR大鼠鼻腔黏膜病理改变程度越低。见插页ⅩⅩⅩⅣ图1C-图1F。
2.4　光学显微镜下各组大鼠鼻腔黏膜组织内嗜酸性粒细胞、中性粒细胞计数比较
与对照组相比，AR模型组大鼠鼻腔黏膜组织内嗜酸性粒细胞及中性粒细胞水平明显升高，差异具有统计学意义（P<0.05）。与模型组相比，鹅不食草提取物不同浓度组及阳性对照组大鼠嗜酸性粒细胞及中性粒细胞水平均有不同程度的下降，差异均具有统计学意义（P<0.05）。且随着鹅不食草提取物浓度的上升，大鼠嗜酸性粒细胞及中性粒细胞水平依次下降，组间两两比较，差异均有统计学意义（P<0.05）。见表3。
2.5　鼻黏膜组织内TLR4及NF-κB相对mRNA及蛋白表达
与对照组相比，AR模型组大鼠鼻腔黏膜组织内TLR4及NF-κB相对mRNA表达量均明显升高，差异具有统计学意义（P<0.05）。与模型组相比，鹅不食草提取物不同浓度组及阳性对照组大鼠TLR4及NF-κB相对mRNA表达量均有不同程度的下降，差异均具有统计学意义（P<0.05）。且随着鹅不食草提取物浓度的上升，大鼠TLR4及NF-κB相对mRNA表达量依次下降，组间两两比较，差异均有统计学意义（P<0.05）。见图2、图3。
2.6　Western-Blot法检测鼻黏膜组织内TLR4及NF-κB蛋白表达量
与对照组相比，AR模型组大鼠鼻腔黏膜组织内TLR4及NF-κB相对蛋白表达量均明显升高，差异具有统计学意义（P<0.05）。与模型组相比，鹅不食草提取物不同浓度组及阳性对照组大鼠TLR4、NF-κB相对蛋白表达量均有不同程度的下降，差异均具有统计学意义（P<0.05）。且随着鹅不食草提取物浓度的上升，大鼠TLR4及NF-κB相对蛋白表达量依次下降，组间两两比较，差异均有统计学意义（P<0.05）。见表4、图4。
2.7　TLR4激动剂对鹅不食草提取物处理组AR大鼠的影响
经TLR4激动剂处理后，鹅不食草提取物不同浓度组AR大鼠鼻腔组织内TLR4、NF-κB相对mRNA表达量均上升，大鼠鼻部症状行为得分为提高，鼻腔黏膜上皮脱落现象加重，黏膜固层中嗜酸性粒细胞及肥大细胞浸润情况加重，鹅不食草提取物组大鼠已好转的AR相关症状及病理改变均向更严重的方向逆转。见表5、图5、图6。
2.8　TLR4激动剂对鹅不食草提取物处理组AR大鼠TLR4、NF-κB蛋白表达的影响
经TLR4激动剂处理后，鹅不食草提取物不同浓度组AR大鼠鼻腔组织内TLR4、NF-κB蛋白及表达量均上升。见表6、图7。
3　讨论
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AR又被称为过敏性鼻炎，主要由特异性致敏原刺激特异性个体所导致的慢性鼻黏膜炎性疾病[14-15]。AR虽不会影响生命安全，但其具有久治难愈、易反复发作的特点，将降低患者生活质量。目前，糖皮质激素是临床上治疗AR最常用的药物，糖皮质激素可直接作用于鼻黏膜，有效改善流涕、鼻痒、打喷嚏等临床症状，但并不能完全根治疾病，且长期使用还将引起鼻部充血、咽部溃疡等不良反应[16-17]。相比而言，中药制剂安全性更高，不会产生用药依赖性，具有较高的临床应用潜能。
鹅不食草在我国分布广泛，主产于江浙、湖北、广西一带，喜欢生长在草地阴凉湿润处，具有辛温解表、止咳解毒、发散风寒的功效，可用于治疗鼻塞、感染、鼻炎等[18]。随着研究技术的不断提高的进展，鹅不食草提取物具有抗炎、抗过敏、抑癌等多种功效。薛鹏辉等[19]研究表明，鹅不食草提取物三萜类化学成分具有良好的抗炎活性。此外，鹅不食草提取物能有效抑制人前列腺癌细胞株体外增殖和侵袭，并可诱导癌细胞凋亡[20]。
本研究采用OVA制作AR模型，并应用不同浓度的鹅不食草提取物处理，结果发现，经鹅不食草提取物处理后，AR大鼠喷嚏、抓鼻及流涕等鼻腔行为得分均明显下降，HE染色病理检查结果显示大鼠鼻腔黏膜组织上皮脱落，以及黏膜固层嗜酸性粒细胞及肥大细胞浸润现象均明显减轻，提示鹅不食草提取物在改善AR大鼠相关临床症状及鼻黏膜病变中具有一定的效果。
Th2型免疫反应是诱导AR的主要原因，包括T细胞、B细胞、嗜酸细胞、嗜碱细胞等多种细胞因子均参与Th2型免疫，共同构成一个复杂的免疫反应网络[21-22]。Th2淋巴细胞被激活后，将促进IgE合成，并增加IL-4、IL-13等炎症因子释放量，IL-4及IL-13是淋巴抗原呈递功能的主要细胞因子，此外，IL-13还参与气道高反应性、嗜酸性粒细胞浸润等病理改变[23-24]。本研究发现，经鹅不食草提取物干预后，AR模型大鼠血清IL-4、IL-13以及IgE水平均较模型组降低，病理组织检查发现，大鼠鼻腔黏膜组织内嗜酸性粒细胞及中性粒细胞水平均降低，提示鹅不食草提取物可通过降低AR大鼠Th2免疫反应，发挥AR治疗功效。
TLR/NF-κB信号通路是重要的细胞免疫通路，其参与免疫应答、炎症反应、细胞凋亡调控等[25]。本研究发现，AR大鼠鼻黏膜组织内TLR4及NF-κB相对mRNA及蛋白表达量均较其对照组上升，提示AR大鼠TLR/NF-κB信号通路被激活。而在使用鹅不食草提取物干预后，AR大鼠鼻黏膜内TLR4及NF-κB相对mRNA及蛋白表达量均下降，且随着鹅不食草提取物浓度的增加，大鼠鼻黏膜内TLR4及NF-κB相对mRNA及蛋白表达量呈依次下降趋势，提示鹅不食草提取物可能通过降低TLR/NF-κB信号通路活性参与AR的治疗。
为验证以上猜想，本研究采用TLR4激动剂LPS处理不同浓度鹅不食草提取物组大鼠，结果显示，经TLR4激动剂LPS处理后，原本经鹅不食草提取物干预后，鼻症状、鼻黏膜病变均减轻的AR大鼠鼻症状、鼻黏膜病变及血清IL-4、IL-13、IgE水平均向严重化方向逆转，证实鹅不食草提取物可通过抑制TLR/NF-κB信号通路，降低Th2型免疫反应，治疗AR。
综上所述，鹅不食草提取物可通过抑制TLR/NF-κB通路，减轻Th2型免疫反应强度，治疗变应性鼻炎。
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2025年第43卷第10期
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doi: 10.13193/j.issn.1673-7717.2025.10.045
  • 首发时间：2026-10-30
  • 出版时间：2025-10-10
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四川省科学技术厅面上项目(2022YFS0629-B4)
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    西南医科大学附属医院，四川 泸州 646000
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Number of
species		 占总种数比例
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total species (%)		 属
Genus		 种数
Number of
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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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