| Purpose:Detection of lung deficiency and phlegm obstruction model rats and normal rats of leukotriene B4 (LTB4), nerve growth factor (NGF), transforming growth factorβ1 (TGF-β1), granulocyte-macrophage colony-stimulating factor (GM-CSF), stem cell factor (SCF) such as the expression of inflammatory factors and to observe the lung deficiency and phlegm obstruction model of rat lungs phlegm after the treatment of Chinese herbal compound the dynamic changes of inflammatory mediators from the perspective of lung deficiency and phlegm with the formation mechanism.Methods:1. Animal groups:male SD rats weighing 180～200g, were randomly assigned to the control group 8, lung deficiency and phlegm syndrome group 14, lung deficiency and phlegm obstruction treated group 14.2. Making touch Methods:lung deficiency and phlegm obstruction model according to the literature (Wang Jiulin, Jiang Wei, Hui-Min Bian. Lung deficiency and phlegm pathological model of development [J]. Traditional Chinese Medicine, 1996,2 (4):44) and improved methods. Rats placed in the morning than the first low growth temperature under 5℃in the cold wind blowing for 10 minutes. Smoked cabinet will be placed in rats, each will be spread evenly 0.2g sulfur powder in half of the root Ai Qing, the fire smoke out animals after 1 hour, every day at a time smoked rats, continuous 42 days. Usually with the normal control group reared in normal environments, feeding normal diet.3. Treatment:modeling is complete, the treatment group Treatment of phlegm to the lungs, according to 10ml/kg/d orally,1 time/day; model group and control group were administered equal volume of normal saline, for 14 days.4. Drawn, staining and image analysis:anesthesia, killed at the groin artery bleeding, thoracotomy, ligation of the left lung bronchus, left lung obtained after washing with normal saline ice, take the middle left lung, placed in 10% formalin fixed, paraffin embedding, slicing, slice thickness 3um, so HE and immunohistochemical staining.2ml saline injection on the right tracheal lung tissue, repeatedly pumping two times, more than 80% recovery rate. The pumping to obtain lavage fluid(BALF) centrifugation(20℃,2000r/min centrifugation 10 min), supernatant collected, frozen standby home-70℃refrigerator.5. LTB4, SCF, TGFβ1 in serum:from lavage fluid by enzyme linked immunosorbent assay (ELISA) test. Specific methods to operate according to kit instructions.6. Lung tissue expression of GM-CSF and NGF detection:Using immunohistochemical method. Specific methods to operate according to kit instructions.7. Statistical analysis:SPSS 16.0 software used to analyze the data to x±s, that the number of each group were compared by F test and t test, P<0.05 as significantly different.Results:1. Animals generallyNormal rats:rats lively, responsive, Maofa Jie Bai Guangze, appetite, no cough, shortness of breath, phlegm and other abnormal signs.Model group:rats activity slow, sometimes cough, sputum, shortness of breath, hair, withered and yellow, mouth and nose secretions increase, food intake and water intake more than normal group.Treated group:Rats in model group than shiny hair, a little cough, sputum.2. Cough AnalysisCough evoked with concentrated ammonia after 3 minutes of normal group(1.30±1.059) in cough frequency of the lowest, with the model group(3.00±1.054) and treatment group(2.30±0.949) were statistically significant differences (P<0.05);model group(3.00±1.054) and treatment group(2.3±0.949) difference was not statistically significant (P>0.05).3. WeightNormal feeding rats a week after the modeling, with modeling of conduct, model group compared with normal weight gain was significantly slowed. Before modeling (first week), the normal group (235.5±6.12) and model group (231.57±9.75) body weight difference was not statistically significant(P>0.05); experiment last week (7th week), the normal group (450.13±60.30), and model group(399.11±41.29) body weight was significantly (P<0.05).4. PathologicNormal rats:trachea, bronchi, ciliated columnar epithelial cells arranged in neat rows cilia regularly arranged, and no significant infiltration of inflammatory cells.Model group:tracheal, bronchial goblet cell hyperplasia, glandular hypertrophy, hyperplasia, mucus secretion exuberant, rare cilia, the lumen filled with a large number of alveolar macrophages, neutrophils, eosinophils and mucus.Treatment group:tracheal, bronchial goblet cell hyperplasia, gland hypertrophy, hyperplasia than those of model group, model group, more than cilia in the lumen of a small amount of alveolar macrophages, neutrophils, eosinophils and mucus.5. LTB4, SCF, TGFβ1 levels of the measured results:LTB4 in BALF in rats of model group (163.944±83.478)>treatment group (156.326±56.388)>normal group (90.820±46.215); SCF model group(0.600±0.244)> treatment group(0.573±0.207)> normal group(0.368±0.116); TGFβ1 model group (391.714±250.639)> treatment group (380.898±312.992)> normal (89.428±61.380), the above indexes in control group and model group, control group and treatment group differences were statistically significant (P<0.05); model group and treatment group difference was not statistically significant(P>0.05).6.Rat lung tissue expression of GM-CSF and NGF test results:Inflammatory mediators in the lung tissue of the percentage of GM-CSF model group (65.710±10.163)> treatment group (51.820±12.505)> control group (46.250±7.440); NGF model group (75.710±13.986)> treatment group (61.820±11.677)> normal group (61.250±16.421), the model group and normal group, treatment group, the percentage of positive cells was significantly (P<0.05), staining signal strength model group(+++) and treatment group (+++) very; Although the normal group and treatment group the percentage of positive cells was no significant difference (P>0.05), but the signal intensity of the treatment group(+++)> stained normal (++).Conclusion:Model group LTB4, NGF, TGF-β1, GM-CSF, SCF level than the treatment group and normal group increases, suggesting that these inflammatory mediators are involved in the pathological process of lung deficiency and phlegm obstruction, but also indicate that lung deficiency and phlegm obstruction of the disease process is a chronic inflammatory process. Lungs phlegm may lower the expression of inflammatory mediators, through its anti-inflammatory effect on the prevention of chronic inflammation play a role. Treatment of Deficient Bufei phlegm phlegm obstruction may be one mechanism of action is through down the expression of inflammatory mediators achieved.
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