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Journal of Chinese Integrative Medicine ›› 2008, Vol. 6 ›› Issue (1): 60-67.doi: 10.3736/jcim20080112

• Original Experimental Research • Previous Articles     Next Articles

A rat model of pulmonary fibrosis induced by infusing bleomycin quickly through tracheal intubation

Wei Zhang1, Xiang-feng Lu2, Xiao-mei Zhang3, Jian-jun Wu4, Liang-duo Jiang1()   

  1. 1. Department of Respiratory Diseases, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China
    2. Department of Pathology, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China
    3. Department of Respiratory Diseases, Dongfang Hospital, Beijing University of Chinese Medicine, Beijing 100078, China
    4. Department of Chinese Medicine, Beijing Royal Integrative Medicine Hospital, Beijing 102209, China
  • Received:2007-04-05 Online:2008-01-20 Published:2008-01-15
  • Contact: JIANG Liang-duo E-mail:liangduojiang@163.com

Objective:To study the approach for developing a rat model of pulmonary fibrosis induced by bleomycin (BLM).

Method:Different doses (7, 6, 5, 3.4, 2, 1 mg/kg) of bleomycin A5-saline were infused into the rats' lung in bleomycin-treated group through tracheal intubation, and rats in sham-operated group were infused with same volume of saline. The living state and lung pathology of the rats were observed. The author deeply studied the condition of the rats in 1 mg/kg bleomycin-treated group, and the changes of body weight and lung pathology were observed. Lung quotient, the content of transforming growth factor β1(TGF-β1)and platelet-derived growth factor (PDGF) in serum were measured on the 14th, 28th and 45th day of the experiment.

Results:The study demonstrated that infusing large doses of bleomycin A5 quickly through tracheal intubation had a high mortality, and infusing 1 mg/kg quickly could successfully develop an animal model of pulmonary fibrosis. Compared with the sham-operated group, fibrosis was appeared obviously in the rats' lung in 1 mg/kg bleomycin A5-treated group after 14 days of experiment, diffuse fibrosis was appeared after 28 days of experiment, and the fibrosis became more severe after 45 days of experiment. The body weight of the rats in bleomycin-treated group was declined after 3, 7 and 14 days of experiment as compared with the sham-operated group (P<0.01). Twenty-one days after the experiment, the body weight was declined too, but there was no significant difference between the bleomycin-treated group and the sham-operated group (P>0.05). Lung quotient was increased 14, 28 and 45 days after the experiment (P<0.01), the level of serum TGF-β1 began to increase since 28 days after the experiment (P<0.05, P<0.01), and the level of serum PDGF also increased gradually 45 days after the experiment (P<0.05). And the mortality rate of 1 mg/kg bleomycin A5-treated group was lower than those of the other doses of bleomycin A5-treated groups.

Conclusion:A rat model of pulmonary fibrosis can be duplicated successfully by infusing 1 mg/kg bleomycin A5 quickly through tracheal intubation.

Key words: pulmonary fibrosis, model, animal, bleomycin, rats

CLC Number: 

  • R521.8

Table 1

Body weight of rats at each time point [$\bar{x}$±s (n), g]"

Group Body weight of rats
Day 0 Day 3 Day 7 Day 14 Day 21 Day 28 Day 35 Day 45
Sham-operated 286±16 (15) 287±20 (15) 319±20 (15) 353±25 (15) 391±33 (10) 421±41 (10) 439±39 (5) 471±45 (5)
BLM-treated 289±15 (25) 268±17 (25)** 285±27 (25)** 316±39 (25)** 362±40 (17) 397±43 (17) 419±51 (9) 448±45 (9)
t 0.542 3.236 4.203 3.275 1.896 1.443 0.786 0.840

Table 2

Lung quotient of rats [$\bar{x}$±s (n)]"

Group Lung quotient of rats
Day 14 Day 28 Day 45
Sham-operated 0.451±0.030 (5) 0.434±0.045 (5) 0.438±0.016 (5)
BLM-treated 0.835±0.264 (8)** 0.726±0.125 (8)** 0.620±0.076 (9)**
t 3.201 5.201 6.914

Table 3

Content of serum TGF-β1 in rats ($\bar{x}$±s, ng/L)"

Group n Content of serum TGF-β1
Day 14 Day 28 Day 45
Sham-operated 5 27.46±5.17 27.62±3.60 27.96±3.69
BLM-treated 8 39.00±14.59 37.76±7.49* 50.73±16.53**
t 2.043 2.797 3.749

Table 4

Content of serum PDGF in rats ($\bar{x}$±s, ng/L)"

Group n Content of serum PDGF
Day 14 Day 28 Day 45
Sham-operated 5 19.52±4.16 18.36±2.76 19.27±8.30
BLM-treated 8 21.56±5.24 23.53±5.82 30.78±6.33*
t 0.731 1.839 2.838

Figure 1

Lung tissue pathology in sham-operated group (HE staining, ×100) The structure of lung was clear. There were no inflammatory cells in alveolar septum, and there were no inflammatory exudates in bronchial lumen and alveolar space."

Figure 2

Lung tissue pathology in sham-operated group (Masson staining, ×100) No pulmonary fibrosis was found."

Figure 3

Lung tissue pathology in BLM-treated group at 24 h of experiment (HE staining, ×100)"

Figure 4

Lung tissue pathology in BLM-treated group at 48 h of experiment (Masson staining, ×100)"

Figure 5

Lung tissue pathology in BLM-treated group on the 7th day of experiment (HE staining, ×100) The inflammation and fibroblast focuses were formed."

Figure 6

Lung tissue pathology in BLM-treated group on the 8th day of experiment (Masson staining, ×100) Deposit of collagen fibers could be seen."

Figure 7

Lung tissue pathology in BLM-treated group on the 14th day of experiment (HE staining, ×100) A great quantity of inflammatory cells could be seen in parts of the lung, and most of them were lymphocytes and eosinophile granulocytes."

Figure 8

Lung tissue pathology in BLM-treated group on the 14th day of experiment (HE staining, ×40) Inequality of cyst-like fiber air space could be seen in parts of the lung, i.e. the formation of honeycomb lung."

Figure 9

Lung tissue pathology in BLM-treated group on the 14th day of experiment (HE staining, ×100) Unequal deposit of collagen fibers in fibrosis area."

Figure 10

Lung tissue pathology in BLM-treated group on the 14th day of experiment (HE staining, ×200) Small amount of inflammatory cells in fibrosis area, and proliferation of typeⅡ alveolar epithelium could be seen."

Figure 11

Lung tissue pathology in BLM-treated group on the 28th day of experiment (HE staining, ×100) Obvious aggregation of macrophages in alveoli."

Figure 12

Lung tissue pathology in BLM-treated group on the 28th day of experiment (HE staining, ×100) Compared with lung tissue pathology on the 14th day of experiment, there were more cyst-like fiber air spaces."

Figure 13

Lung tissue pathology in BLM-treated group on the 14th day of experiment (Masson staining, ×100) Obvious fibrosis could be seen in parts of the lung."

Figure 14

Lung tissue pathology in BLM-treated group on the 28th day of experiment (Masson staining, ×100) The pulmonary fibrosis was more serious than that on the 14th day of experiment."

Figure 15

Lung tissue pathology in BLM-treated group on the 45th day of experiment (HE staining, ×200) Less inflammatory cells in fibrosis area and quantity of fibroblasts and collagen fibers could be seen."

Figure 16

Lung tissue pathology in BLM-treated group on the 45th day of experiment (Masson staining, ×200) The fibrosis was more serious than that on the 28th day of the experiment."

[1] American Thoracic Society/European Respiratory Society International Multidisciplinary Consensus Classification of the Idiopathic Interstitial Pneumonias. This joint statement of the American Thoracic Society (ATS) , and the European Respiratory Society (ERS) was adopted by the ATS board of directors, June 2001 and by the ERS Executive Committee, June 2001. Am J Respir Crit Care Med. 2002; 165(2):277-304.
doi: 10.1164/ajrccm.165.2.ats01
[2] Yi XH. ATS/ERS classification and pathologic diagnosis of idiopathic interstitial pneumonia. Zhen Duan Bing Li Xue Za Zhi. 2005; 12(6):401-404. Chinese.
doi: 10.3969/j.issn.1007-8096.2005.06.001
易祥华 . 介绍特发性间质性肺炎的ats/ers分类及病理诊断.诊断病理学杂志. 2005; 12(6):401-404.
doi: 10.3969/j.issn.1007-8096.2005.06.001
[3] Adamson IY, Bowden DH . The pathogenesis of bleomycin-induced pulmonary fibrosis in mice. Am J Pathol. 1974; 77(2):185-197.
pmid: 4141224
[4] Thall RS, Barton RW, Damato DA , et al. Differential cellular analysis of bronchoalveolar lavage fluid obtained at various stages during the development of bleomycin- induced pulmonary fibrosis in the rat. Am Rev Respir Dis. 1982; 126(3):488-492.
[5] Ludwicka A, Ohha T, Trojanowska M , et al. Elevated levels of platelet derived growth factor and transforming growth factor-beta 1 in bronchoalveolar lavage fluid from patients with scleroderma. J Rheumatol. 1995 ; 22(10):1876-1883.
[6] Yoshida M, Sakuma J, Hayashi S , et al. A histologically distinctive interstitial pneumonia induced by overexpression of the interleukin 6, transforming growth factor beta 1,or platelet-derived growth factor B gene. Proc Natl AcadSciUSA. 1995; 92(21):9570-9574.
doi: 10.1073/pnas.92.21.9570
[7] Sime PJ, Xing Z, Graham FL , et al. Adenovector- mediated gene transfer of active transforming growth factor-beta 1 induces prolonged severe fibrosis in rat lung. J Clin Invest. 1997; 100(4):768-776.
[8] Lee CG, Cho SJ, Kang MJ , et al. Early growth response gene 1-mediated apoptosis is essential for transforming growth factor beta 1-induced pulmonary fibrosis. J Exp Med. 2004; 200(3):377-389.
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