朝医麻黄定喘汤对OVA诱导的过敏性哮喘豚鼠模型HMGB1/TLR4/NF-κB信号通路的影响  

Effects of Chao medicine Mahuang-Dingchuan decoction on HMGB1/TLR4/NF-κB signaling pathway in guinea pig model of allergic asthma induced by OVA

作　　者：朴颖 王重阳 宋艺兰 孟庆玲 郑明昱 金丽娜 李良昌 延光海 PIAO Ying;WANG Chong-yang;SONG Yi-lan;MENG Qing-ling;ZHENG Ming-yu;JIN Li-na;LI Liang-chang;YAN Guang-hai(Key Laboratory of Immunization and Targeted Research on Common Allergic Diseases in Jilin Province,Yanji 133002,China;Department of Emergency,Affiliated Hospital of Yanbian University,Yanji 133000,China;Department of Anato-my,School of Medicine,Yanbian University,Yanji 133002,China;Department of Korean Medicine,School of Medicine,Yanbian University,Yanji 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]延边大学附属医院急诊科,吉林延吉133000 [3]延边大学医学院解剖学教研室,吉林延吉133002 [4]延边大学医学院朝医教研室,吉林延吉133002

出　　处：《中国病理生理杂志》2022年第5期913-919,共7页Chinese Journal of Pathophysiology

基　　金：吉林省科技厅医药健康领域重点研发项目(No.20200404096YY)。

摘　　要：目的:探讨朝医麻黄定喘汤对过敏性哮喘豚鼠气道炎症的影响,并探讨其作用机制是否与HMGB1/TLR4/NF-κB信号通路有关。方法:选取50只雄性豚鼠随机分为正常对照组、模型组、低剂量(5 g/kg)麻黄定喘汤组、高剂量(20 g/kg)麻黄定喘汤组和地塞米松(0.5 mg/kg)组,每组各10只。除正常对照组外,其余4组均采用卵清蛋白(OVA)腹腔注射致敏和OVA气溶胶雾化激发制备哮喘豚鼠模型。最后一次雾化后观察豚鼠行为学改变;末次激发24 h后处死豚鼠,采集支气管肺泡灌洗液(BALF),通过Diff-Quik染色进行炎症细胞计数;取肺组织标本,HE、PAS和Masson染色观察肺组织病理学变化;ELISA法检测BALF中IL-4、IL-5和IL-13含量;流式细胞术测定小鼠肺组织中IL-4和IFN-γ水平;Western blot检测肺组织中HMGB1、TLR4、MyD88和NF-κB蛋白表达;免疫组织化学及免疫荧光检测豚鼠肺组织中HMGB1表达。结果:麻黄定喘汤能有效减轻OVA诱导的哮喘豚鼠气道炎症,抑制杯状细胞增生和胶原纤维沉积,减少BALF中IL-4、IL-5和IL-13含量,并在降低肺组织中IL-4水平的同时升高肺组织中IFN-γ水平;下调肺组织HMGB1、TLR4、MyD88和NF-κB蛋白表达,减弱肺组织HMGB1荧光强度。结论:麻黄定喘汤能够有效减轻哮喘豚鼠气道炎症,可能与调节HMGB1/TLR4/NF-κB信号通路有关。AIM:To investigate the effect of Chao medicine Mahuang-Dingchuan decoction on airway inflammation in guinea pigs with allergic asthma and its potential mechanisms.METHODS:Fifty male guinea pigs were randomly divided into normal control group,model group,low-dose(5 g/kg)Mahuang-Dingchuan decoction group,high-dose(20 g/kg)Mahuang-Dingchuan decoction group and dexamethasone(0.5 mg/kg)group,with 10 guinea pigs in each group.The allergic asthma model was etablished by intraperitoneal injection of ovalbumin(OVA)and atomizing inhalation of OVA aerosol in the latter 4 groups.The behavioral changes of the guinea pigs were observed after atomization for the last time.The guinea pigs were sacrificed 24 h after the final treatment,and then bronchoalveolar lavage fluid(BALF)was collected.Inflammatory cells in the BALF were recorded by Diff-Quik staining.HE staining,PAS staining and Mas son staining were used to observe lung histopathological changes.The levels of IL-4,IL-5 and IL-13 in the BALF were mesured by ELISA.Western blot was used to detect the protein expression levels of HMGB1,TLR4,MyD88 and NF-κB in lung tissues.Immunohistochemistry and immunofluorescence were used to detect the expression of HMGB1 in guinea pig lung tissues.RESULTS:Mahuang-Dingchuan decoction attenuated OVA-induced airway inflammation in asthmatic guinea pigs,inhibited goblet cell proliferation and collagen fiber deposition,reduced the levels of IL-4,IL-5,and IL-13 in the BALF,and decreased the level of IL-4 in lung tissues,but increased the level of IFN-γin lung tissues.Mahuang-Dingchuan decoction reduced the protein expression of HMGB1,TLR4,MyD88 and NF-κB,and the fluorescence intensity of HMGB1 in guinea pig lung tissues.CONCLUSION:Mahuang-Dingchuan decoction reduces airway inflammation in asthmatic guinea pigs through HMGB1/TLR4/NF-κB signaling pathway.

关 键 词：麻黄定喘汤 过敏性哮喘 气道炎症 HMGB1/TLR4/NF-κB信号通路 

分 类 号：R285.5[医药卫生—中药学] R562[医药卫生—中医学]