基于TGF-β1/Smad2/3信号通路探讨茴香胶囊抗支气管哮喘作用及机制

Anti-bronchial asthma effect of Huixiang capsules based on TGF-β1/Smad2/3 signaling pathway and related mechanism

目的探讨茴香胶囊抗支气管哮喘作用及潜在机制。方法采用环磷酰胺腹腔注射法建立小鼠免疫低下模型评价茴香胶囊免疫增强作用;采用浓氨水引咳法及酚红排泄法评价茴香胶囊止咳祛痰活性;采用卵清蛋白致敏和激发方式构建支气管哮喘小鼠模型,实验过程中观察并记录小鼠体重及行为学变化;检测小鼠肺泡灌洗液(BALF)中炎症细胞计数;ELISA法检测免疫球蛋白E(IgE)、γ-干扰素(IFN-γ)、白细胞介素-4(IL-4)、白细胞介素-17A(IL^(-1)7A)、转化生长因子-β1(TGF-β1)水平;HE和Masson染色观察肺组织病理学变化;RT-qPCR、Western blot检测TGF-β1/Smad2/3通路相关蛋白表达。结果茴香胶囊可增强免疫低下小鼠的免疫功能,延长小鼠咳嗽潜伏期,减少小鼠咳嗽次数,增加小鼠气管酚红排泌量,降低支气管哮喘小鼠BALF中白细胞等炎性细胞数量,降低IgE、IL-4、IL^(-1)7A、TGF-β1水平,增加IFN-γ水平,减轻小鼠肺组织病理学变化;下调TGF-β1/Smad2/3通路相关蛋白表达。结论茴香胶囊可能通过增强机体免疫力、止咳祛痰缓解支气管哮喘小鼠哮喘症状,调节Th1/Th2失衡、降低肺组织中炎症因子水平、下调TGF-β1/Smad2/3信号通路减少气道炎症及气道重塑。

Objective To determine the anti-bronchial asthma effect and related mechanism of Huixiang capsules based on immune enhancement and cough expectoration.Methods Cyclophosphamide intraperitoneal injection was used to establish a mouse immunocompromised model to evaluate the immune enhancing effect of Huixiang capsules.Concentrated ammonia cough inducing method and phenol red excretion method were used to evaluate the cough expectorant activity of Huixiang capsules.Ovalbumin sensitization and excitation were used to establish a mouse model of bronchial asthma.The body weight of mice and behavioral changes were observed and recorded in the experiment.The inflammatory cells were counted in the BALF of the mice.IgE,IFN-γ,IL-4,IL^(-1)7A,and TGF-β1 were detected by ELISA.Pathological changes in the lung tissues were observed by HE and Masson staining.The levels of TGF-β1,Smad2,Smad3,α-SMA,and VEGF mRNA expressions were determined by RT-qPCR and Western blot.Results Huixiang capsule enhanced the immune function in immunocompromised mice,prolonged the cough latency,reduced the cough times,and lowered the amount of phenol red excretion in the trachea.The number of inflammatory cells such as white blood cells in the BALF of mice with bronchial asthma was reduced as well.The levels of IgE,IL-4,IL^(-1)7A,and TGF-β1 were decreased,while the IFN-γlevel was increased.The pathological changes in the lungs of mice were relieved and the expression of TGF-β1/Smad2/3 pathway related proteins down-regulated.Conclusion Huixiang capsules may alleviate symptoms in mice with bronchial asthma by enhancing the immunity,relieving cough and expectorant,regulating Th1/Th2 imbalance,decreasing inflammatory factor levels in the lung tissues,and down-regulating TGF-β1/Smad2/3 signaling pathway to reduce airway inflammation and remodel the airway.