实验动物与比较医学 ›› 2026, Vol. 46 ›› Issue (5): 668-676.DOI: 10.12300/j.issn.1674-5817.2026.001

• 人类疾病动物模型 • 上一篇    下一篇

一种痰湿证慢性阻塞性肺疾病大鼠模型的建立与评价

张攀攀1,2, 许金森2, 程佑民3, 吴华嵩3, 林静瑜1,2()()   

  1. 1. 福建省中医药科学院药物研究所, 福州 350003
    2. 福建省中医药科学院经络感传重点实验室, 福州 350003
    3. 福建省中医药科学院实验动物中心, 福州 350003
  • 收稿日期:2026-01-03 修回日期:2026-06-11 出版日期:2026-10-25 发布日期:2026-10-16
  • 通讯作者: 林静瑜(1973—),女,博士,副研究员,研究方向:针药结合的效应机制研究。E-mail: jingyul2012@163.com。ORCID: 0009-0001-4292-1129
  • 作者简介:
    张攀攀(1994—),女,硕士研究生,研究方向:针灸的效应机制研究。E-mail: 1397002257@qq.com
  • 基金资助:
    国家自然科学基金“基于人工智能辅助分析探讨循经感传与针刺靶向趋病的关系”(82274413); 福建省科技计划项目“卤地菊对脓毒症急性肺损伤的保护作用及其NLRP3炎症小体机制”(2022R1003006)

Establishment and Evaluation of a Rat Model of Chronic Obstructive Pulmonary Disease with Phlegm-Dampness Syndrome

ZHANG Panpan1,2, XU Jinsen2, CHENG Youmin3, WU Huasong3, LIN Jingyu1,2()()   

  1. 1. Institute of Materia Medica, Fujian Academy of Chinese Medical Sciences, Fuzhou 350003, China
    2. Key Laboratory of Propagated Sensation Along Meridian, Fujian Academy of Chinese Medical Sciences, Fuzhou 350003, China
    3. Institute of Laboratory Animal Science, Fujian Academy of Chinese Medical Sciences, Fuzhou 350003, China
  • Received:2026-01-03 Revised:2026-06-11 Published:2026-10-25 Online:2026-10-16
  • Correspondence to:
    LIN Jingyu (ORCID:0009-0001-4292-1129), E-mail:

摘要:

目的 建立一种符合中医痰湿证的慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)病证结合模型,并进行表型评价。 方法 将18只SPF级8周龄雄性SD大鼠,随机分为对照组(n=8)和模型组(n=10)。造模耗时19周,对照组无干预。造模方法:分别于第1、5、9、13、17周(滴注周)的起始日上午9:00,向大鼠行气管内滴注脂多糖(lipopolysaccharide,LPS),共5次,每次给药剂量为40 μg/只,当日给药1次。除滴注周外,每日于自制高湿环境的烟熏箱内,让大鼠接受香烟烟雾暴露(第9周开始前15支香烟/次,自第9周起20支香烟/次,每日30 min/次)。每周测量一次大鼠体重,观察其一般体征及痰湿证表征。造模结束次日麻醉大鼠,以进行肺通气功能测定。采用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测血清、支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和黏蛋白5AC(mucin 5AC,MUC5AC)的含量;对肺组织进行HE染色和阿尔辛蓝-过碘酸希夫染色(alcian blue and periodic acid-Schiff staining,AB-PAS),以观察肺组织形态结构、杯状细胞的增生情况;采用蛋白质印迹法检测肺组织中TNF-α、MUC5AC蛋白的表达量。 结果 体征与一般状态方面,与对照组比较,模型组大鼠体重自第3周起均显著减轻(P<0.01),并出现毛色发黄、活动迟缓、懒动、呼吸喘促及口鼻分泌物增多等现象。肺通气功能方面,与对照组比较,模型组大鼠肺潮气量、每分通气量均显著下降(P<0.01),动态顺应性显著降低(P<0.05),而肺弹性阻力显著增加(P<0.01)。炎症因子和黏液相关蛋白方面,与对照组比较,模型组大鼠的血清、BALF和肺组织中TNF-α、MUC5AC蛋白水平均显著增高(P<0.01)。病理形态学方面,HE染色结果显示,模型组大鼠肺泡明显扩张、破裂并部分融合成肺大疱,细支气管腔内有炎性细胞浸润;AB-PAS染色结果显示,模型组大鼠细支气管黏膜上皮杯状细胞增多,腔内形成黏液栓。 结论 本造模方法可复制出符合中医痰湿证症状且具有肺通气功能下降与肺气肿、炎症、慢性支气管炎样病理特征的COPD大鼠模型,可为后续中医药治疗痰湿证COPD研究提供可靠支撑。

关键词: 慢性阻塞性肺疾病, 痰湿证, 病证结合模型, 肿瘤坏死因子-α, 黏蛋白5AC, SD大鼠

Abstract:

Objective To establish and evaluate a rat model of chronic obstructive pulmonary disease (COPD) associated with the traditional Chinese medicine (TCM) phlegm-dampness syndrome. Methods Eighteen SPF-grade 8-week-old male SD rats were randomly assigned to a control group (n=8) and a model group (n=10). The modeling procedure lasted for 19 weeks. Rats in the control group received no intervention. Lipopolysaccharide (LPS; 40 μg per rat per instillation) was instilled intratracheally into rats in the model group at 09:00 on the first day of weeks 1, 5, 9, 13, and 17, for a total of five instillations. Except for the instillation weeks, rats were exposed daily to cigarette smoke in a self-made high-humidity smoke exposure chamber (15 cigarettes/session before week 9 and 20 cigarettes/session from week 9 onward; 30 min/session/day). Body weight was recorded weekly, and general signs and manifestations of TCM phlegm-dampness syndrome were observed. On the day after modeling, rats were anesthetized for pulmonary ventilatory function testing. Tumor necrosis factor-α (TNF-α) and mucin 5AC (MUC5AC) levels in serum and bronchoalveolar lavage fluid (BALF) were measured using enzyme-linked immunosorbent assay (ELISA). Lung tissues were subjected to hematoxylin and eosin (HE) and alcian blue and periodic acid-Schiff staining (AB-PAS) to assess lung tissue morphology and goblet cell hyperplasia. Western blotting was performed to determine TNF-α and MUC5AC protein expression in lung tissue. Results General signs and overall condition: Compared with the control group, body weight in the model group was significantly decreased from week 3 onward (P<0.01). Rats showed yellowish fur, sluggish activity, reduced movement, rapid/labored breathing, and increased oral/nasal secretions. Pulmonary ventilatory function: Compared with the control group, minute ventilation and tidal volume (both P<0.01) and dynamic compliance (P<0.05) were decreased, whereas elastic resistance was markedly increased (P<0.01) in the model group. Inflammatory factors and mucus-related proteins: Compared with the control group, TNF-α and MUC5AC levels in serum and BALF, as well as their protein levels in lung tissue, were significantly elevated in the model group (all P<0.01). Histopathology: HE staining showed obvious alveolar dilatation and rupture with partial coalescence into bullae, and inflammatory cell infiltration in bronchiolar lumens. AB-PAS staining revealed increased goblet cells in bronchiolar mucosal epithelium and mucus plug formation in the lumen. Conclusion This protocol can reproduce a COPD rat model with manifestations consistent with TCM phlegm-dampness syndrome, reduced pulmonary ventilation function, and pathological features of emphysema, inflammation, and chronic bronchitis, thereby providing reliable support for subsequent studies on TCM treatment of COPD with phlegm-dampness syndrome.

Key words: Chronic obstructive pulmonary disease, Phlegm-dampness syndrome, Disease-syndrome combination model, Tumor necrosis factor-α, Mucin 5AC, SD rat

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