祛风通窍止咳方对上气道咳嗽综合征模型豚鼠咳嗽敏感性及肺组织、鼻黏膜组织TRPV4及神经源性炎症的影响  

The Effects of Qufeng Tongqiao Cough-Relieving Decoction (祛风通窍止咳方) on Cough Sensitivity,TRPV4 in Lung and Nasal Mucosal Tissues,and Neurogenic Inflammation in a Guinea Pig Model of Upper Airway Cough Syndrome

作　　者：罗景舒 马建岭 史利卿 季坤 刘松 范钰晗 李先莉 郭兆迪 LUO Jingshu;MA Jianling;SHI Liqing;JI Kun;LIU Song;FAN Yuhan;LI Xianli;GUO Zhaodi(Beijing University of Chinese Medicine,Beijing,100029;Dongfang Hospital,Beijing University of Chinese Medicine)

机构地区：[1]北京中医药大学,北京市100029 [2]北京中医药大学东方医院

出　　处：《中医杂志》2025年第5期518-525,共8页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(82474472);2023年北京中医药大学研究生自主科研课题(ZJKT2023096)。

摘　　要：目的 探讨祛风通窍止咳方治疗上气道咳嗽综合征(UACS)的可能作用机制。方法 24只豚鼠随机分为空白组、模型组、中药组、抑制剂组,每组6只。除空白组外,其余各组豚鼠采用腹腔注射卵清蛋白+氢氧化铝致敏后卵清蛋白滴鼻联合烟熏的方法构建UACS豚鼠模型。造模后中药组予祛风通窍止咳方0.9 g/(100 g·d)灌胃,抑制剂组予瞬时受体电位香草酸亚型4(TRPV4)抑制剂GSK2193874 1 mmol/L雾化,每天1次,每次5 min。空白组、模型组给予2 ml/(100 g·d)生理盐水灌胃,每日1次。给药7天后,以0.4 mol/L柠檬酸进行咳嗽激发试验;ELISA检测血清中IgE水平及血清、肺泡灌洗液、鼻腔灌洗液中炎症因子[包括白细胞介素6 (IL-6)、白细胞介素8 (IL-8)]水平;HE染色观察肺组织及鼻黏膜组织病理改变;免疫组化法检测肺组织、鼻黏膜组织TRPV4及神经源性炎症因子[包括P物质(SP)、降钙素基因相关肽(CGRP)]蛋白水平;实时荧光定量聚合酶链式反应法(Real-time PCR)检测肺组织TRPV4、SP、CGRP mRNA表达。结果 HE染色结果显示,模型组肺组织及鼻黏膜组织结构破坏明显、炎症细胞浸润,而中药组和抑制剂组肺组织、鼻黏膜组织病理损伤有所改善。与空白组比较,模型组咳嗽次数、血清IgE水平及血清、肺泡灌洗液、鼻腔灌洗液中IL-6、IL-8水平增加,肺组织、鼻黏膜组织中TRPV4、SP、CGRP蛋白水平及其mRNA表达升高(P<0.05或P<0.01);与模型组比较,中药组和抑制剂组咳嗽次数、血清IgE水平及肺组织TRPV4、SP mRNA表达降低,中药组血清、肺泡灌洗液、鼻腔灌洗液中IL-6、IL-8水平降低,肺组织、鼻黏膜组织中TRPV4、CGRP蛋白水平降低,抑制剂组血清IL-6、IL-8水平降低,肺泡灌洗液中IL-6水平降低,鼻腔灌洗液中IL-8水平降低,肺组织TRPV4、SP、CGRP蛋白水平及鼻黏膜组织SP、CGRP蛋白水平降低(P<0.05或P<0.01)。与中药组比较,抑制剂组血清IgE、IL-6、IL-8水平�Objective To investigate the potential mechanism of action of the Qufeng Tongqiao Cough-relieving Decoction(祛风通窍止咳方,QTCD)in the treatment of upper airway cough syndrome(UACS).Methods Twenty-four guinea pigs were randomly divided into blank group,model group,traditional Chinese medicine(TCM)group,and inhibitor group,with six guinea pigs in each group.Except for the blank group,guinea pigs were sensitized with ovalbumin and aluminum hydroxide via intraperitoneal injection,followed by ovalbumin nasal drops combined with smoke exposure to establish the UACS model.After modeling,the TCM group was administered QTCD 0.9 g/(100 g·d)by gavage,the inhibitor group received the transient receptor potential vanilloid receptor 4(TRPV4)inhibitor GSK21938741 mmol/L,5 min by nebulisation,and the blank group and model group were given 2 ml/(100 g·d)normal saline by gavage once daily.After 7 days of treatment,a cough provocation test was performed using 0.4 mol/L citric acid.The levels of IgE in serum and inflammatory cytokines,including interleukin-6(IL-6),interleukin-8(IL-8)in serum,bronchoalveolar lavage fluid(BALF),and nasal lavage fluid(NLF)were detected by enzyme-linked immunosorbent assay(ELISA).Histopathological changes in lung and nasal mucosal tissues were observed by hematoxylin-eosin(HE)staining.Immunohistochemistry was used to detect the protein levels of TRPV4,substance P(SP),and calcitonin gene-related peptide(CGRP)in lung and nasal mucosal tissues.Real-time polymerase chain reaction(Real-time PCR)was used to detect the mRNA expression of TRPV4,SP,and CGRP in lung tissues.Results HE staining showed significant structural damage and infiltration of inflammatory cells in the lung and nasal mucosal tissues in the model group,while the TCM group and inhibitor group showed improved pathological changes.Compared with the blank group,the model group showed increased cough frequency,serum IgE level,and IL-6 and IL-8 levels in serum,BALF,and NLF.The protein levels of TRPV4,SP,and CGRP in lung and nasal mucosal

关 键 词：上气道咳嗽综合征 咳嗽敏感性 瞬时受体电位香草酸亚型4 神经源性炎症 祛风通窍止咳方 

分 类 号：R285.5[医药卫生—中药学] R-332[医药卫生—中医学]