痰瘀互结证哮喘大鼠模型的建立及评价  

Establishing and evaluating of asthma rat model with phlegm and blood stasis syndrome

作　　者：袁欣 王安琪 王思宇[1] 潘乐芯 汪佳颖 邹璐[1] 杨爱东[1,2] YUAN Xin;WANG Anqi;WANG Siyu;PAN Lexin;WANG Jiaying;ZOU Lu;YANG Aidong(School of Traditional Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Center for Traditional Chinese Medicine and Epidemic Disease,Shanghai Institute of InfectiousDisease and Biosecurity,Shanghai 200032,China)

机构地区：[1]上海中医药大学中医学院,上海201203 [2]上海市重大传染病和生物安全研究院中医药疫病研究中心,上海200032

出　　处：《中国实验动物学报》2024年第10期1243-1251,共9页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金(82104708);上海市市级科技重大专项(ZD2021CY001,ZXS004R4-1)。

摘　　要：目的探索痰瘀互结证哮喘大鼠模型的建立及评价方法。方法60只SD雄性大鼠随机分为5组(正常组、哮喘模型组、病证结合组、地塞米松组、咳喘六味合剂组),每组12只。通过卵清蛋白(ovalbumin,OVA)诱导建立哮喘疾病模型;通过高脂饮食联合冰水浴法建立痰瘀互结证动物模型。通过动物行为学观察、病理切片观察、炎症指标检测、气道反应性测定评估哮喘疾病模型;通过体重测定,证候观察,血糖、血脂、凝血功能、血液流变学指标检测,以及咳喘六味合剂干预后效果观察评估痰瘀互结证模型。结果(1)OVA诱导后哮喘模型组、病证结合组大鼠均出现呼吸急促,张口喘息,腹部翕动,烦躁不安,易激惹等现象;苏木素-伊红(HE)染色见肺组织支气管黏膜上皮排列紊乱,局部脱落,基底膜增厚,支气管管壁增厚,管腔狭窄,大量炎性细胞浸润,毛细血管充血;血清中血清中白细胞介素4(interleukin 4,IL-4)、白细胞介素6(interleukin 6,IL-6)、转化生长因子-β1(transformating growth factor betal,TGF-β1)较正常组均上升(P<0.05);各浓度乙酰甲胆碱(methacholine,Mch)激发后较正常组均表现出气道阻力(enhance pause,Penh)值增高(P<0.05)。(2)造模结束后,病证结合组大鼠出现寒颤、蜷缩少动、爪甲紫暗,尾部可见紫黑瘀斑、便溏,肛周不洁等证候表现;与哮喘模型组比较,病证结合组大鼠体重上升(P<0.05);血糖、甘油三酯、总胆固醇含量升高(P<0.05),凝血酶时间缩短(P<0.05),纤维蛋白原含量增多(P<0.05),低、中、高切变率下全血黏度均升高(P<0.05);咳喘六味合剂组以上各项指标较病证结合组有改善。结论通过OVA诱导及高脂饮食联合冰水浴法建立痰瘀互结证哮喘大鼠模型,通过行为学观察、指标检测、以方测证综合评价该病证结合模型具有可行性。Objective To explore method of establishing and evaluating an asthma rat model with phlegm and blood stasis syndrome.Methods 60 SD male rats were randomly divided into 5 groups,a normal group,asthma group,combination of disease and syndrome(combination)group,DM group,and KCLW group,with 12 rats in each group.Asthma models were established using ovalbumin(OVA).A syndrome model of phlegm and blood stasis was established using a high-fat diet combined with the ice water bath method.We evaluated the asthma model through animal behavior observation,pathological section observation,inflammation index detection,and airway reactivity measurements.The phlegm and blood stasis syndrome model was evaluated via measurements of rat body mass,blood glucose,blood lipids,coagulation function,and hemorheological indexes and by observing symptoms and syndrome determination by Kechuan Liuwei mixture.Results(1)After OVA induction,the rats in the asthma model group and combination group showed symptoms such as shortness of breath,open mouth breathing,abdominal movement,restlessness,and irritability.HE staining showed the disordered arrangement of the bronchial mucosa in lung tissue,local detachment,thickening of the basement membrane and the bronchial tube wall,narrowing of the lumen,extensive infiltration of inflammatory cells,and congestion of capillaries.Compared with the normal group,the asthma model group and combination group(P<0.05)had increased serum IL-4,IL-6,and TGF-β1.Penh values were increased after stimulation with various concentrations of Mch(P<0.05).(2)Rats in the combination group showed symptoms such as chills,curling up with minimal movement,purple and dark claws,purple and black bruises on the tail,loose stools,and unclean perianal area.Compared with the rats in the asthma model group,rats in the combination group had increased body mass(P<0.05)and blood glucose,triglyceride,and total cholesterol levels(P<0.05),a shortened thrombin time(P<0.05),increased fibrinogen content(P<0.05),and significantly increased whole

关 键 词：支气管哮喘 痰瘀互结证 病证结合 动物模型 以方测证 

分 类 号：Q95-33[生物学—动物学]