摘要
目的 探究麻黄碱对支气管哮喘小鼠气道重塑的影响及对转化生长因子β1(TGF-β1)/Smads通路的调控作用。方法 将所有小鼠随机分为对照组、模型组、地塞米松组、麻黄碱低剂量组、麻黄碱高剂量组、麻黄碱高+TGF-β1激活剂组,每组12只。除对照组,其余组小鼠腹腔注射含有40μg卵清蛋白(OVA)和2 mg 10%氢氧化铝的致敏液诱导哮喘模型。8周后,检测各组小鼠气道高反应性;对支气管肺泡灌洗液(BALF)中嗜酸性粒细胞进行计数;ELISA检测BALF中白细胞介素(IL)-4、IL-5和IL-13的水平;苏木精-伊红(HE)染色和Masson染色观察小鼠肺组织的病理学变化及胶原纤维面积;免疫组织化学检测小鼠肺组织中α-平滑肌肌动蛋白(α-SMA)表达;Western blot检测小鼠肺组织中TGF-β1/Smads通路相关蛋白的表达。结果 与对照组相比,模型组小鼠不同乙酰甲胆碱剂量下的气道高反应性、气道壁的总面积(Wat)/基底膜的周长(Pbm)比值、胶原纤维面积、嗜酸性粒细胞及IL-4、IL-5、IL-13含量、α-SMA、TGF-β1表达及p-Smad2/Smad2、p-Smad3/Smad3比值均升高(P<0.05),Smad7表达降低(P<0.05);与模型组相比,地塞米松组和麻黄碱低、高剂量组小鼠不同乙酰甲胆碱剂量下的气道高反应性、Wat/Pbm比值、胶原纤维面积、嗜酸性粒细胞及IL-4、IL-5、IL-13含量、α-SMA、TGF-β1表达及p-Smad2/Smad2、p-Smad3/Smad3比值均降低(P<0.05),Smad7表达升高(P<0.05);与麻黄碱高剂量组相比,麻黄碱高+TGF-β1激活剂组小鼠不同乙酰甲胆碱剂量下的气道高反应性、Wat/Pbm比值、胶原纤维面积、嗜酸性粒细胞及IL-4、IL-5、IL-13含量、α-SMA、TGF-β1表达及p-Smad2/Smad2、pSmad3/Smad3比值均升高(P<0.05),Smad7表达降低(P<0.05)。结论 麻黄碱可以改善支气管哮喘小鼠的气道重塑,其机制与调控TGF-β1/Smads信号通路有关。
Objective To explore the effect of ephedrine on airway remodeling and its regulation effect on transforming growth factor-β1(TGF-β1)/Smads pathway in asthmatic mice.Methods All mice were randomly separated into control group,model group,dexamethasone group,ephedrine low-dose group,ephedrine high-dose group,and ephedrine high-dose+TGF-β1 activator group,with 12 mice in each group.Except for the control group,mice in other groups were intraperitoneally injected with 40μg ovalbumin(OVA)and 2 mg 10%aluminium hydroxide sensitizer to induce asthma models.After 8 weeks,the airway hyperresponsiveness of mice in each group was detected;eosinophils in bronchoalveolar lavage fluid(BALF)were counted;ELISA was applied to detect the levels of interleukin(IL)-4,IL-5,and IL-13 in BALF;hematoxylin-eosin(HE)staining and Masson staining were applied to observe the pathological changes and collagen fiber area of mouse lung tissue;immunohistochemistry was applied to detect the expression ofα-smooth muscle actin(α-SMA)protein in mouse lung tissue;Western blot was applied to detect the expression of TGF-β1/Smads pathway related proteins in mouse lung tissue.Results Compared with the control group,the airway hyperreactivity at different doses of methacholine,total area of airway wall(Wat)/perimeter of the basement membrane(Pbm)ratio,collagen fiber area,eosinophil,the contents of IL-4,IL-5,IL-13,the expression ofα-SMA,TGF-β1,the ratios of p-Smad2/Smad2 and p-Smad3/Smad3 in the model group increased(P<0.05),the expression of Smad7 decreased(P<0.05);compared with the model group,the airway hyperreactivity at different doses of methacholine,Wat/Pbm ratio,collagen fiber area,eosinophil,the contents of IL-4,IL-5,IL-13,the expression ofα-SMA,TGF-β1,the ratios of p-Smad2/Smad2 and p-Smad3/Smad3 in the dexamethasone group,the ephedrine low-dose and high-dose groups decreased(P<0.05),the expression of Smad7 increased(P<0.05);compared with the ephedrine high-dose group,the airway hyperreactivity at different doses of methacholine,Wat/Pbm ratio,collagen fiber area,eosinophil,the contents of IL-4,IL-5,IL-13,the expression ofα-SMA,TGF-β1,the ratios of p-Smad2/Smad2 and p-Smad3/Smad3 in the ephedrine high-dose+TGF-β1 activator group increased(P<0.05),the expression of Smad7 decreased(P<0.05).Conclusion Ephedrine can improve airway remodeling in asthmatic mice,and its mechanism is related to the regulation of TGF-β1/Smads signaling pathway.
作者
范慧慧
任玉梅
田新磊
张凯
李晓丽
Fan Huihui;Ren Yumei;Tian Xinlei;Zhang Kai;Li Xiaoli(Dept of Pediatrics,Henan Hospital of Traditional Chinese Medicine,the Second Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000;College of Pediatrics,Henan University of Chinese Medicine,Zhengzhou 450000)
出处
《安徽医科大学学报》
CAS
北大核心
2024年第8期1398-1404,共7页
Acta Universitatis Medicinalis Anhui
基金
河南省中医药科学研究专项(编号:20-21ZY2057)。
