摘要
目的探讨搜风愈喘方(Soufengyuchuan Recipe,SFYCR)及其拆方干预哮喘大鼠Th1/Th2免疫失衡的作用机制,并探究“祛风”“化瘀”“化痰”3种治法干预哮喘气道炎症的疗效。方法将70只大鼠按随机数字表分为对照组、模型组、地塞米松组、消暗瘀方组、祛宿痰方组、祛伏风方组、搜风愈喘方组,共7组,每组10只。除空白对照组外其余各组大鼠予卵清白蛋白(OVA)注射和雾化复制哮喘大鼠模型。末次给药12 h后,麻醉大鼠并取材,苏木精-伊红(HE)、过典酸雪夫氏(PAS)染色法观察肺组织病理形态学变化及杯状细胞增生情况,酶联免疫吸附测定(ELISA)法检测血清及肺泡灌洗液中免疫球蛋白E(IgE)、白细胞介素-4(IL-4)、白细胞介素-13(IL-13)、干扰素γ(IFN-γ)含量,酶联免疫吸附法(PCR)检测肺组织匀浆中GATA结合蛋白3(GATA-3)、T盒子转录因子(T-bet)的基因表达。蛋白质印迹法(Western blot)检测肺组织中GATA-3、T-bet蛋白的表达。结果与对照组比较,模型组大鼠气道平滑肌增厚,支气管黏膜上皮细胞坏死、脱落,并伴有大量炎症细胞浸润,IL-13、IL-4、IgE、GATA-3表达增多(P<0.05),IFN-γ、T-bet表达下降(P<0.05)。地塞米松组及各中药组病理染色中的气道炎症程度均较模型组降低,IL-13、IL-4、IgE、GATA-3表达较模型组减少(P<0.05),IFN-γ、T-bet表达较模型组增多(P<0.05)。各中药组组间比较表明,搜风愈喘方全方及祛伏风方在改善气道炎症、降低IL-13、IL-4、IgE、GATA-3因子水平方面优于其余拆方组。结论搜风愈喘方及拆方可减少哮喘大鼠IL-13、IL-4、IgE、GATA-3的表达,升高IFN-γ、T-bet表达,改善Th1/Th2免疫失衡,“祛风”“化瘀”“化痰”3种治法相比较,“祛风”法在改善气道炎症方面疗效较优。
Objective To explore the mechanism of Soufeng yuchuan Recipe(搜风愈喘方,SFYCR)and its disassembled prescriptions on the immune imbalance of Th1/Th2 in asthmatic rats and explore the therapeutic effects of“dispelling wind”,“resolving blood stasis”and“resolving phlegm”on airway inflammation of asthma.Methods According to the random number table,70 rats were divided into 7 groups:blank control group,model group,dexamethasone group,SFYCR group,dispellling latent wind(Qufufeng,QFF)group,resolving blood stasis(Xiaoanyu,XAY)group and eliminating abiding phlegm(Qusutan,QST)group,with 10 rats in each group.Except the blank control group,the rats in other groups were sensitized by mixed injection of ovalbumin(OVA)and aluminum hydroxide and challenged by aerosol inhalation to replicate the rat model of asthma.Twelve hours after the last administration,the anesthetized rats were taken out,and the pathomorphological changes of lung tissue were observed by HE and Masson staining.The contents of immunoglobulin E(IgE),interleukin-4(IL-4),interleukin-13(IL-13)and interferon-γ(IFN-γ)in serum and bronchoalveolar lavage fluid were detected by ELISA.The gene expressions of GATA binding protein 3(GATA-3)and T box transcription factor(T-bet)in lung tissue homogenate were detected by enzyme linked immunosorbent assay(PCR).Western blotting was used to detect the expressions of GATA-3 and T-bet in lung tissue.Results Compared with those of the blank control group,the airway smooth muscle of the model group was thickened,the bronchial mucosal epithelial cells were necrotic and exfoliated,accompanied by a large number of inflammatory cell infiltration,the expressions of IL-13,IL-4,IgE and GATA-3 increased,while the expressions of IFN-γand T-bet decreased.The degree of airway inflammation in the dexamethasone group and each TCM group was lower than that in the model group,and the expressions of IL-13,IL-4,IgE and GATA-3 were lower than those in the model group,while the expressions of IFN-γand T-bet were higher than those in the model group.According to the comparison between the disassembled prescription groups,the QFF group was superior to the other groups in improving airway pathology and reducing the levels of IL-13,IL-4,IgE and GATA-3.Conclusion SFYCR and its disassembled formulae can reduce the expressions of IL-13,IL-4,IgE and GATA-3 in asthma rats and increase expressions of IFN-γand T-bet to improve Th1/Th2 immune imbalance.Among the three treatment methods of“dispelling wind”,“resolving blood stasis”and“resolving phlegm”,the“dispelling wind method”has better curative effect in improving airway inflammation.
作者
张宇婧
梁磊
郭旭冉
李牧瑶
陈晨
闫永彬
ZHANG Yujing;LIANG Lei;GUO Xuran;LI Muyao;CHEN Chen;YAN Yongbin(College of Pediatrics,Henan University of Chinese Medicine,Zhengzhou 450000,Henan,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,Henan,China)
出处
《中华中医药学刊》
CAS
北大核心
2023年第11期188-192,I0037,I0038,共7页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金面上项目(81973903,82174438)
河南省中医药科学研究专项(20-21ZY1002)。
关键词
免疫失衡
哮喘
伏风暗瘀宿痰
搜风愈喘方
immune imbalance
asthma
latent wind-blood stasis-abiding phlegm
Soufeng yuchuan Recipe(搜风愈喘方)
