中药蓝玉簪颗粒对慢性哮喘小鼠气道重建的影响及机制

Efficacy of a Chinese herb medicine,Lanyuzan granules,on airway remo-delling in a murine model of chronic asthma and its mechanism

目的:观察中药蓝玉簪颗粒对慢性哮喘小鼠气道重建的抑制作用并探讨其机制.方法:将32只雌性BABL/c小鼠随机分成正常对照组(A组),慢性哮喘模型组(B组),蓝玉簪颗粒组(C组,1.0g生药/3.0kg体质量)和地塞米松干预组(D组,2.0mg/kg),以卵白蛋白(OVA)腹腔注射致敏并长期雾化吸入制备小鼠慢性哮喘模型.苏木素-伊红(HE)染色观察小鼠肺组织一般病理学改变;阿尔辛蓝-过碘酸-希夫氏(AB-PAS)染色观察气道上皮杯状细胞增生、黏液分泌情况;天狼猩红(SR)染色观察气道壁胶原沉积并检测肺组织羟脯氨酸(HYP)含量以评价气道上皮下纤维化,采用免疫组化半定量法测定气道壁转化生长因子β1(TGF-β1)含量.计算机图像分析软件测定气道外、内径比值(Po/Pi),气道壁厚度和面积(Awt,Aaw)以及TGF-β1免疫组化阳性染色面积与气道基底膜周径(Pbm)比值(如Awt/Pbm,TGF-β1/Pbm)等.结果:B组小鼠气管及血管周围大量炎性细胞浸润,气道黏膜上皮增生,气道黏液高分泌及气道上皮下胶原沉积增多,气道壁TGF-β1表达增强;Po/Pi,Awt/Pbm,Aaw/Pbm以及TGF-β1/Pbm,肺组织HYP含量等所有指标均显著增高.C组与B组之间比较差异均有统计学意义(P<0.01);C组与A,D组比较差异具有统计学意义(P<0.05).TGF-β1的表达与肺组织HYP含量呈显著正相关(rs=0.669,P<0.01).结论:中药蓝玉簪颗粒可以抑制慢性哮喘小鼠气道炎症及气道重建,其机制可能与抑制TGF-β1表达有关.

AIM： To evaluate the protective effect of Lanyuzan granules on airway remodelling in a murine model of chronic asth- ma and its mechanism. METHODS： Female BABL/C mice were randomly divided into 4 groups （n- 8 in each group） ： the normal group （A）, the model group （B）, the Lanyuzan granules group （ C, 1.0 g/3.0 kg body mass） and the dexamethasone group （ D, 2.0 mg/kg body mass）. The chronic asthmatic model was estab- lished with the ovalbumin sensitization and repeated inhalation of ovalbumin aerosols. Lung tissue sections were stained with hema- toxylin and eosin （HE） for general morphology, with Alician Blue-Periodic Acid-Schiff （AB-PAS） for identification of goblet cells, with Sirius Red stain for determination of collagen accumu- lation and hydroxyproline content to evaluate the airway fibrosis, and immunostained for assessment of transforming growth factor β1 （TGF-β1）. We calculated the changes of airway remodelling dimensions, such as the airway outer perimeter （Po）, the inter- nal perimeter （Pi）, airway wall thickness （Awt）, airway wall area （Awa） and the area of positive staining of TGF-β1 content in the airway wall with the computerized image analysis system （Image Pro-Plus 6.0, Media Cybernetics, USA）. Finally, all dimensions were referenced to the perimeter of the basement mem- brane （Pbm） （e.g. Aaw/Pbm, TGF-β1/Pbm）. RESULTS： Mice exposed to recurrent airway challenge with ovalbumin had characteristics of airway inflammation and airway remodeling, in-cluding subepithelial fibrosis, goblet cell hyperplasia and mucus hypersecretion, and airway wall thickening. The differences of all the airway remodelling dimensions and the others dimensions, such as the positive staining of TGF-β1/Pbm, and the hydroxyproline content between B and C groups were significant （P 〈 0.01 ）; differences were also significant when group C was compared with group A and group D （P 〈 0.05）. A close correla- tion between TGF-β1 and the hydroxyproline content was demon- strated （rs = 0. 669, P 〈 0.01 ）. CONCLUSION： Lanyuzan granules might suppress airway remodeling and airway inflamma- tion by decreasing the expression of TGF-β1.