慢性阻塞性肺疾病肺脾气虚证大鼠模型的建立及评价  

Establishment and Evaluation of Rat Model for Chronic Obstructive Pulmonary Disease with Lung-spleen Qi Deficiency

作　　者：丁焕章 吴迪[1] 杨勤军 许浩燃 慈慧敏 吴凡[1] 童佳兵[1,2,3,4] 高雅婷 朱洁 李泽庚[1,2,3,4,5] DING Huanzhang;WU Di;YANG Qinjun;XU Haoran;CI Huimin;WU Fan;TONG Jiabing;GAO Yating;ZHU Jie;LI Zegeng(Anhui University of Chinese Medicine,Hefei 230038,China;The First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,China;Institute of Respiratory Disease Prevention and Treatment of Chinese Medicine,Anhui Academy of Chinese Medicine,Hefei 230031,China;Anhui Province Key Laboratory of Application and Transformation of Traditional Chinese Medicine in Prevention and Treatment of Major Pulmonary Diseases,Hefei 230031,China;Key Laboratory of Xin'an Medicine,Ministry of Education,Hefei 230038,China)

机构地区：[1]安徽中医药大学,合肥230038 [2]安徽中医药大学第一附属医院,合肥230031 [3]安徽省中医药科学院中医呼吸病防治研究所,合肥230031 [4]中医药防治肺系重大疾病应用转化安徽省重点实验室,合肥230031 [5]新安医学教育部重点实验室,合肥230038

出　　处：《中国实验方剂学杂志》2023年第23期47-55,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金联合重点项目(U20A20398);全国名中医肺系疾病学术思想传承及特色方药的临床开发研究项目(202204295107020045);安徽省科技重大专项(201903a07020015);安徽高校自然科学研究项目(KJ2021A0572)。

摘　　要：目的:建立并评价慢性阻塞性肺疾病(COPD)肺脾气虚病证结合模型。方法:采用“香烟烟熏法+脂多糖(LPS)气管滴注联合番泻叶浸液灌胃”构建COPD肺脾气虚证大鼠模型。将40只SPF级SD雄性大鼠随机分成空白组、模型组、番泻叶低、中、高剂量病证结合模型组,共5组。除空白组外各组均每日烟熏,并于第1天、第14天气管内滴注LPS。番泻叶低、中、高剂量病证结合模型组在造模第28天予4℃不同剂量番泻叶浸液(5、10、20 g·kg^(-1))灌胃3周,造模共49 d。采集宏观指标:一般情况(体质量、摄食量、粪便含水量、肛温)及行为学指标(抓力测试、鼠尾悬挂实验)。测定肺功能,肺组织病理,血清D-木糖、淀粉酶、胃泌素含量,肺泡灌洗液白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)水平,外周血T淋巴细胞亚群(CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+))水平,计算胸腺指数、脾指数。结果:一般情况方面,番泻叶高剂量组大鼠死亡2只。与空白组比较,番泻叶中、高剂量组体质量、摄食量均显著降低(P<0.01),粪便含水量显著升高(P<0.01),番泻叶高剂量组肛温较空白组显著降低(P<0.01)。行为学指标方面,与空白组比较,各造模组大鼠抓力均显著降低(P<0.01);番泻叶中、高剂量组静止不动时间明显增加(P<0.05,P<0.01)。肺功能方面,与空白组比较,各造模组大鼠第0.3秒用力呼气容积(FEV0.3)、FEV0.3/用力肺活量(FVC)均显著降低(P<0.01)。肺组织苏木素-伊红(HE)染色可见各造模组大鼠支气管壁增厚,杯状细胞增生,部分肺泡融合呈肺气肿改变。胃肠功能指标方面,与空白组比较,番泻叶中、高剂量组D-木糖、胃泌素、α-淀粉酶水平均显著降低(P<0.01)。免疫炎症指标方面,与空白组比较,番泻叶中、高剂量组脾、胸腺指数均显著减小(P<0.01),各造模组大鼠外周血CD4^(+)水平均显著降低(P<0.01),CD4^(+)/CD8^(+)明显下降(P<0.05,P<0.01),肺泡灌洗液IL-1β�Objective:To establish and evaluate a chronic obstructive pulmonary disease(COPD)model with lung-spleen qi deficiency.Method:A rat model mimicking COPD with lung-spleen qi deficiency was established by the combination of cigarette smoking and intratracheal instillation of lipopolysaccharide(LPS)along with gavage of Sennae Folium infusion.Forty male SPF-grade SD rats were randomly assigned to blank,model,and low-(L-FXY),medium-(M-FXY),and high-dose(H-FXY)Sennae Folium infusion groups.Other groups except the blank group were exposed to daily cigarette smoke,with LPS administrated via intratracheal instillation on the 1st and 14th days.On the 28th day of modeling,the L-FXY,M-FXY,and HFXY groups were administrated with Sennae Folium infusion at 5,10,and 20 g·kg^(-1),respectively,and at 4℃for three weeks.The modeling lasted for 49 days.The general conditions(body mass,food intake,fecal water content,and anal temperature)and behaviors(grip strength test and tail suspension test)of rats in different groups were examined.The lung function,lung histopathology,D-xylose,amylase,and gastrin levels in the serum,interleukin(IL)-1βand IL-6 levels in the alveolar lavage fluid,levels of T-lymphocyte subsets(CD4^(+),CD8^(+),and CD4^(+)/CD8^(+))in the peripheral blood,and thymus and spleen indices were measured.Result:Two rats died in the H-FXY group.Compared with the blank group,both the M-FXY and H-FXY groups exhibited reduced body mass and food intake(P<0.01)and increased fecal water content(P<0.01).The anal temperature in the H-FXY group was lower than that in the blank group(P<0.01).The grip strength decreased in the modeling groups compared with the blank group(P<0.01),and the duration of immobility in the tail suspension test increased in the M-FXY and H-FXY groups(P<0.05,P<0.01).Compared with the blank group,the modeling groups showed reduced 0.3 second forced expiratory volume(FEV0.3),FEV0.3/forced vital capacity(FVC)(P<0.01),thickening of bronchial walls,proliferation of goblet cells,and the presence of emphysematous

关 键 词：慢性阻塞性肺疾病 肺脾气虚证 动物模型 病证结合模型 模型评价 

分 类 号：R2-0[医药卫生—中医学] R33R289R318.13