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

doi:10.12300/j.issn.1674-5817.2026.001

Abstract

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

CLC Number:

R-332
R563.9

ZHANG Panpan,XU Jinsen,CHENG Youmin,et al. Establishment and Evaluation of a Rat Model of Chronic Obstructive Pulmonary Disease with Phlegm-Dampness Syndrome[J]. Laboratory Animal and Comparative Medicine, 2026, 46(5): 668-676. DOI: 10.12300/j.issn.1674-5817.2026.001.

Figures/Tables 6

Table 1

Traditional Chinese medicine syndrome scoring scale for rats with phlegm-dampness syndrome in chronic obstructive pulmonary disease models

评分

Score

大鼠行为

Rat behavior

精神状态

Mental state

毛发、耳、尾

Hair, ear, tail

舌苔

Tongue coating

饮食、体重

Diet and body weight

Figure 1 Comparison of body weight gain changes between the two groups and smoke exposure status in rats

Note： A， Body weight gain line chart of rats in the two groups （ $x ¯$ ±s，n=8）. Compared with the control group， ^* P<0.05， ^** P<0.01. B， Gross appearance comparison between the two groups of rats （Photographs of rats at week 19 of the experiment）. C， Status of rats in the model group during smoke exposure.

Figure 1 Comparison of body weight gain changes between the two groups and smoke exposure status in rats

Figure 2 Comparison of pulmonary dynamic compliance， elastic resistance， minute ventilation， and tidal volume between the two groups of rats

Note： A， Pulmonary dynamic compliance in rats； B， Lung elastic resistance in rats； C， Minute ventilation volume in rats； D， Tidal volume in rats. n=6 rats per group. Compared with the control group， ^* P<0.05， ^** P<0.01. n=6.

Figure 3 Comparison of TNF-<strong>α</strong> and MUC5AC levels in serum and BALF between the two groups of rats

Note： A， Comparison of tumor necrosis factor-α （TNF-α） levels in rat serum； B， Comparison of mucin 5AC （MUC5AC） levels in rat serum； C， Comparison of TNF-α levels in rat bronchoalveolar lavage fluid （BALF）； D， Comparison of MUC5AC levels in rat BALF. Model group （n=8）. Compared with the control group， ^** P<0.01.

Figure 4 Comparison of the morphological structure of lung tissue between two groups of rats

Note： A， Hematoxylin and eosin （HE） staining of lung tissue （scale bars from left to right are 250 μm， 100 μm， 100 μm， and 50 μm）； B， Alcian blue and periodic acid-Schiff staining （AB-PAS） of lung tissue （scale bars from left to right are 100 μm， 50 μm， 100 μm， and 50 μm）.

Figure 5 Comparison of TNF-<strong>α</strong> and MUC5AC protein expres-sion in lung tissue between the two groups of rats

Note： A， Representative Western blot bands； B， Relative TNF-α protein expression in lung tissue； C， Relative mucin 5AC （MUC5AC） protein expression in lung tissue. n=8 rats per group. Compared with the control group， ^** P<0.01.

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