朝医鹿茸大补汤对咳嗽变异性哮喘豚鼠TLR4/WNT-5A信号通路的影响  

Effects of traditional korean medicine Lurong Dabu Decoction on TLR4/WNT-5A signaling pathway in guinea pigs with cough variant asthma

作　　者：李慧文 金丽娜 宋艺兰 李良昌 延光海[1,3] 孟庆玲 李欢[1,3] 刘凯月 郑明昱 LI Hui-wen;JIN Li-na;SONG Yi-lan;LI liang-chang;YAN Guang-hai;MENG Qing-ling;LI-Huan;LIU Kai-yue;ZHENG Ming-yu(Key Laboratory of Allergic Diseases of Jilin Provincial Science and Technology Agency,Yanji,Jilin 133002,China;Yanbian University Medical College,Yanji,Jilin 133002,China;Dept of Neonatology,Affiliated Hospital of Yanbian University,Yanji,Jilin 133002,China)

机构地区：[1]吉林省过敏性常见疾病免疫与靶向研究重点实验室,吉林延吉133002 [2]延边大学附属医院新生儿科,吉林延吉133002 [3]延边大学医学院,吉林延吉133002

出　　处：《中国药理学通报》2023年第5期978-985,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 82060911)。

摘　　要：目的探讨鹿茸大补汤对咳嗽变异性哮喘的作用机制。方法将豚鼠随机分为正常组(CON)、模型组(OVA)、鹿茸大补汤高剂量组(HIGH)、鹿茸大补汤低剂量组(LOW)、地塞米松组(DEX)。通过烟熏加注射卵清蛋白(OVA)、氢氧化铝溶液及雾化吸入激发咳嗽来创建咳嗽变异性哮喘(cough variant asthma,CVA)模型。最后一次刺激豚鼠后24 h内解剖,取其肺泡灌洗液(BALF)和肺组织。利用免疫吸附(ELISA)法测定BALF中炎症细胞种类和炎性细胞因子的含量;取左肺中叶,进行HE染色和Masson染色,观察病理变化;用免疫组织化学染色法观察肺部组织中TLR4和WNT-5A蛋白的表达分布;从左肺上叶提取蛋白做免疫蛋白印迹(Western blot),测肺组织中TLR4、WNT-5A蛋白水平;用免疫荧光法检测肺组织中TLR4、WNT-5A荧光强度;采用流式细胞术在豚鼠肺组织中检测IL-4和IFN-γ。结果鹿茸大补汤能够改善豚鼠气道炎症反应,抑制胶原纤维沉积,降低BALF中IL-4、IL-5、IL-13含量,抑制肺组织中TLR4、WNMT-5A的蛋白表达,升高肺组织中的IFN-γ水平,同时降低IL-4水平。结论鹿茸大补汤可能通过TLR4/WNT-5A信号通路抑制CVA的发生。Aim To discuss the mechanism of Lurong Dabu Decoction on cough variant asthma.Methods Guinea pigs were divided into normal group(CON),model group(OVA),Lurong Dabu Decoction high-dose group(HIGH),low-dose group(LOW),and dexamethasone group(DEX)at random.The CVA model was established by smoking plus injection of OVA,aluminum hydroxide solution and nebulized inhalation to stimulate cough.Gguinea pigs were dissected 24 hours after the last challenge to obtain alveolar lavage fluid(BALF)and lung tissues.Immunoadsorption(ELISA)method was applied to detect the types of inflammatory cells and the content of inflammatory cytokines in BALF;HE and Masson staining of the middle lobe of the left lung were used to observe the pathological changes in lung tissues;immunohistochemical staining was used to observe TLR4 and WNT-5A protein expression and distribution of lung tissues;the protein extracted from the upper lobe of the left lung was used to measure the level of TLR4 and WNT-5A protein in lung tissues by Western blot;immunofluorescence was employed to measure the fluorescence intensity of TLR4 and WNT-5A in lung tissues;flow cytometry was used to detect IL-4 and IFN-γin guinea pig lung tissues.Results Lurong Dabu Decoction could improve guinea pig airway inflammation,inhibit collagen fiber deposition,reduce the content of IL-4,IL-5,and IL-13 in BALF,and inhibit the protein expression of TLR4 and WNT-5A in lung tissues and increase IFN-γlevels in lung tissues while decreasing IL-4 levels.Conclusion Lurong Dabu Decoction may inhibit the occurrence of CVA through TLR4/WNT-5A signaling pathway.

关 键 词：朝医 鹿茸大补汤 咳嗽变异性哮喘 TLR4 WNT-5A 豚鼠 

分 类 号：R-332[医药卫生] R282.74R322.35R441.5R562.25