咳嗽变异性哮喘风伏阴伤证大鼠模型建立与评价  

Establishment and Evaluation of Rat model of Cough Variant Asthma with Syndrome of Latent Wind Damaging Yin

作　　者：于明霞 兰瑞恒 余思洋 沈佳锋 张志杰 张诗瑜 姜博涵 崔红生 YU Mingxia;LAN Ruiheng;YU Siyang;SHEN Jiafeng;ZHANG Zhijie;ZHANG Shiyu;JIANG Bohan;CUI Hongsheng(The Third Affiliated Hospital of Beijing University of Chinese Medicine,Beijing 100029,China;Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学第三附属医院,北京100029 [2]北京中医药大学,北京100029

出　　处：《中国实验方剂学杂志》2024年第16期100-109,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82274497)。

摘　　要：目的:探讨咳嗽变异性哮喘(CVA)风伏阴伤证大鼠模型的建立与评价。方法:60只大鼠随机分为空白组、CVA风伏阴伤组、CVA组、布地奈德组、桑梅止咳组,每组12只。除空白组外,各组大鼠采用卵蛋白氢氧化铝腹腔注射致敏+卵蛋白氢氧化铝雾化激发致敏,其中CVA风伏阴伤组,布地奈德组和桑梅止咳组在此基础上给予“吹风”和“甲状腺素片”灌胃(0.15 g·kg^(-1)·d^(-1))制备咳嗽变异性哮喘风伏阴伤证模型。实验第1~15天为模型制备阶段,第16~29天为以方测证阶段。第16天,布地奈德组给予雾化吸入治疗(0.5 g·L^(-1)),桑梅止咳组给予颗粒剂灌胃(4.19 g·kg^(-1)),各组每天治疗1次,持续14 d。于第1、8、15天,比较空白组、CVA风伏阴伤组、CVA组大鼠的体质量、饮水量、肛温、心前区及四肢爪心的温度的变化,实验第28天,比较大鼠体质量、饮水量、肛温、心前区及四肢爪心的温度的变化,第29天,进行咳嗽敏感性测定,辣椒素(1×10^(-4) mol·L^(-1))引咳,记录大鼠2 min内咳嗽次数,第30天,测定肺功能指标,检测血中炎症因子白细胞介素(IL)-4、IL-5、IL-13、血清干扰素(IFN)-γ及免疫球蛋白E(IgE)指标,检测血清中环磷酸腺苷(cAMP)、环磷酸鸟苷(cGMP)含量;苏木素-伊红(HE)染色观察肺组织病理,实时荧光定量聚合酶链式反应(Real-time PCR)检测肺组织cAMP、IL-4、IL-5、IL-13相对表达量。结果:第15天,与空白组和CVA组比较,CVA风伏阴伤组体质量降低,饮水量明显增加,肛温明显增高,心前区及四肢爪心温度均升高(P<0.05);第28天,与CVA风伏阴伤组比较,布地奈德组、桑梅止咳组饮水量减少,肛温降低,心前区及四肢爪心温度明显降低(P<0.05),与布地奈德组比较,桑梅止咳组心前区及左下、右下肢爪心温度降低(P<0.05)。在咳嗽敏感性方面,与空白组比较,CVA风伏阴伤组和CVA组2 min内咳嗽次数明显增多(P<0.05),与CVA风伏阴伤组比较,桑梅�Objective:To investigate the establishment and evaluation of a rat model of cough variant asthma(CVA,syndrome of latent wind damaging Yin).Method:Sixty rats were randomized into Blank,CVA+latent wind damaging Yin,CVA,budesonide,and Sangmei Zhike granules,with 12 rats in each group.The rats in other groups except the blank group were sensitized by intraperitoneal injection of ovalbumin and aluminum hydroxide+atomization inhalation of ovalbumin and aluminum hydroxide.The rats in the groups of CVA+latent wind damaging Yin,budesonide,and Sangmei Zhike granules were then placed in a draught and administrated with thyroxine tablets(0.15 g·kg^(-1))by gavage to induce latent wind damaging Yin.Days 1-15 of the experiment constituted the modeling stage while days 16-29 served as the verification stage.On day 16,the rats in the budesonide group received atomization inhalation of the drug(0.5 g·L^(-1)),and those in the Sangmei Zhike granules group were administrated with the granules(4.19 g·kg^(-1))by gavage once a day for 14 days.On days 1,8,and 15,the body mass,water intake,anal temperature,and temperatures in the precordium and paw center were compared among the blank,CVA+latent wind damaging Yin,and CVA groups.On day 28,the body mass,water intake,anal temperature,and temperatures in the precordium and paw center were compared among five groups.On day 29,cough sensitivity was assessed based on the number of capsaicin(1×10^(-4) mol·L^(-1))-induced coughs within 2 min.On day 30,the pulmonary function indexes and the serum levels of inflammatory mediators[interleukin(IL)-4,IL-5,IL-13,interferon(IFN)-γ,and total immunoglobulin E(IgE)],cyclic adenosine monophosphate(cAMP),and cyclic guanosine monophosphate(cGMP)were determined.Hematoxylin-eosin staining was employed to observe the pathological damage in the lung.Real-time PCR was carried out to determine the relative expression levels of cAMP,IL-4,IL-5,and IL-13 in the lung tissue.Result:On day 15,compared with the blank and CVA groups,the CVA+latent wind damaging Yin gr

关 键 词：咳嗽变异性哮喘 风伏阴伤 动物模型 以方测证 病证结合 

分 类 号：R2-0[医药卫生—中医学] R22R242R285.5R563