主 办:北 京 中 医 药 大 学
ISSN 2095-6606 CN 10-1157/R

现代中医临床 ›› 2021, Vol. 28 ›› Issue (1): 44-50.doi: 10.3969/j.issn.2095-6606.2021.01.010

• 中医临床基础研究 • 上一篇    下一篇

慢性咳嗽寒饮伏肺病证结合动物模型的建立与评价*

王丽云1, 史利卿1, 王颖2, 马建岭1, 李扭扭1, 罗敬月3, 温绍惠1, 季坤1#   

  1. 1 北京中医药大学东方医院 北京 100078;
    2 浙江省中医院;
    3 首都医科大学附属北京胸科医院
  • 收稿日期:2020-06-05 出版日期:2021-01-30 发布日期:2021-02-02
  • 通讯作者: #季坤,女,博士,副主任医师,E-mail:jikun004@126.com
  • 作者简介:王丽云,女,硕士,主治医师
  • 基金资助:
    *国家自然科学基金资助项目(No.81373588),北京市自然科学基金面上项目(No.7192115),北京市自然科学基金青年项目(No.7184224),中央高校基本科研业务费专项资金资助(No.2018-JYBZZ-JS144)

Establishment and evaluation of an animal model with chronic cough with cold fluid retained in the lung*

Wang Liyun1, Shi Liqing1, Wang Ying2, Ma Jianling1, Li Niuniu1, Luo Jingyue3, Wen Shaohui1, Ji Kun1#   

  1. 1 Dongfang Hospital, Beijing University of Chinese Medicine, Beijing 100078;
    2 Zhejiang Hospital of Traditional Chinese Medicine;
    3 Beijing Chest Hospital, Capital Medical University
  • Received:2020-06-05 Online:2021-01-30 Published:2021-02-02

摘要: 目的 探讨慢性咳嗽寒饮伏肺病证结合动物模型的构建方法,并进行可靠性评价。方法 将48只雄性SPF健康豚鼠随机分为正常对照组、咳嗽模型组、形寒饮冷组、病证模型组,每组12只。采用卵蛋白致敏,建立咳嗽敏感性增高动物模型,结合形寒饮冷刺激,建立寒饮伏肺中医证候模型,通过两者结合建立慢性咳嗽寒饮伏肺病证结合模型。观察豚鼠一般状态与行为,咳嗽次数,气道阻力,肺组织病理,肺组织瞬时受体电位锚蛋白1(TRPA1)、P物质(SP)、降钙素基因相关肽(CGRP)mRNA表达及肺组织TRPA1、P物质受体(NK-1R)、CGRP蛋白表达。结果 与正常对照组、咳嗽模型组比较,形寒饮冷组、病证模型组豚鼠的一般状态与行为均出现寒饮伏肺证的症状表现,其中病证模型组更明显。与正常对照组比较,咳嗽模型组、病证模型组咳嗽次数均增加,气道阻力均升高,肺组织TRPA1,SP,CGRP mRNA表达均升高,肺组织TRPA1,NK-1R,CGRP蛋白表达均升高(P<0.01),形寒饮冷组肺组织TRPA1蛋白表达升高(P<0.01)。与咳嗽模型组比较,形寒饮冷组咳嗽次数减少,气道阻力均降低,肺组织TRPA1,SP,CGRP mRNA表达均降低,肺组织TRPA1,NK-1R,CGRP蛋白表达均降低(P<0.01);病证模型组咳嗽次数增加,气道阻力均升高,肺组织TRPA1,SP,CGRP mRNA表达均升高,肺组织TRPA1,NK-1R,CGRP蛋白表达均升高(P<0.01或P<0.05)。与形寒饮冷组比较,病证模型组咳嗽次数增加,气道阻力均升高,肺组织TRPA1,SP,CGRP mRNA表达均升高,肺组织TRPA1,NK-1R,CGRP蛋白表达均升高(P<0.05或P<0.01)。与正常对照组比较,其余3组肺组织病理均可见肺组织结构破坏及炎症反应,病变程度由低到高依次为形寒饮冷组、咳嗽模型组、病证模型组。结论 采用中西医复合病因干预方法可成功建立慢性咳嗽寒饮伏肺病证模型。该模型既符合西医发病机制,又体现中医证候特征,可为慢性咳嗽的机制探讨及中药药效机理研究提供较为可靠的模型依据。

关键词: 慢性咳嗽, 寒饮伏肺证, 动物模型, 豚鼠

Abstract: Objective To discuss the establishment method of disease and syndrome-integrated animal model of chronic cough with cold fluid retained in the lung, and evaluate its reliability. Methods 48 male SPF guinea pigs were randomly divided into the normal control group, the cough model group, the cold body and diet group, and disease syndrome combination model group (n=12 in each group). The cough hypersensitivity animal model of was established by ovalbumin(OVA), while the traditional Chinese medicine (TCM) syndrome model of cold fluid retained in the lung was established by cold body and cold drink stimulation. Therefore, the disease and syndrome combination animal model of chronic cough with cold fluid retained in the lung was established by combining the above two methods. The general state and behaviors, cough frequency, airway resistance, lung pathology, the mRNA expression of lung transient receptor potential ankyrin A1(hereafter TRPA1), substance P(hereafter SP), calcitonin gene-related peptide(hereafter CGRP) and the protein expression of lung TRPA1, neurokinin-1 receptor (hereafter NK-1R) and CGRP were observed and measured. Results As compared with the normal control group and the cough model group, symptoms of chronic cough with cold fluid retained in the lung including the general state and behaviors of the guinea pigs were more obvious in the cold body and diet group and especially in the disease syndrome combination model group. As compared with the normal control group, the cough frequency, airway resistance, the mRNA expression of lung TRPA1, SP and CGRP, the protein expression of lung TRPA1, NK-1R and CGRP were all elevated in the cough model group and the disease syndrome combination model group (P<0.01), and the protein expression of lung TRPA1 was significantly increased in the cold body and diet group (P<0.01). As compared with the cough model group, cough frequency, airway resistance, the mRNA expression of lung TRPA1, SP and CGRP, the protein expression of lung TRPA1, NK-1R and CGRP were all significantly decreased in the cold body and diet group(P<0.01) and significantly increased in the disease syndrome combination model group(P<0.01 or P<0.05). As compared with cold body and diet group, cough frequency, airway resistance, the mRNA expression of lung TRPA1, SP and CGRP, the protein expression of lung TRPA1, NK-1R and CGRP were all significantly increased in the disease syndrome combination model group(P<0.01 or P<0.05). As compared with the normal control group, lung pathology showed apparent structural destruction and tissue inflammation in the other three groups. The pathological severity from low to high were the cold body and diet group, the cough model group and the disease syndrome model. Conclusions The disease and syndrome combination animal model of chronic cough with cold fluid retained in the lung could be successfully established by the TCM and Western interventions. This model not only conforms to the pathogenesis of Western medicine, but also reflects the characteristics of TCM syndromes. It can provide a reliable model basis for the study of the mechanism of chronic cough and the pharmacodynamics in TCM.

Key words: chronic cough, cold fluid retained in lung, animal model, guinea pig

中图分类号: 

  • R285.6