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The Effect of Buqihuoxue Recipe on Chronic Bronchitis with Lung-QI Deficiency Syndrome Rats’ TH1 and TH2 Related Cytokines

Author: HuRui
Tutor: FangChaoYi
School: Hebei Medical University
Course: Diagnostics of Chinese Medicine
Keywords: Chronic bronchitis, lung deficiency IL-4 IFN-γ TH1/TH2 Cytokines
CLC: R259
Type: Master's thesis
Year: 2009
Downloads: 137
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Abstract

Purpose: chronic bronchitis (CB) is a common disease of the respiratory system, a tracheobronchial mucosa as well as one of the surrounding tissue non-specific inflammation, mainly for cough, expectoration, or accompanied by wheezing and recurrent episodes. With the population aging and atmospheric pollution, the incidence of chronic bronchitis increasingly more serious impact on people's health. Qi Deficiency Syndrome chronic bronchitis syndrome. In recent years, many scholars of modern technological means, more extensive research from many aspects of chronic bronchitis, and that chronic bronchitis analyzed from the perspective of inflammation, and can be summarized as promoting inflammatory cytokines and anti-inflammatory factor system imbalance caused bronchial inflammation. Promote secretion of inflammatory cytokines and anti-inflammatory cytokines sources about can be divided into three categories: T cells secrete, secretion of monocyte-macrophage cells, neutrophils (PMN) secretion. TH1 and TH2 cells are further differentiated by the T cells from the secretion of pro-inflammatory cytokines and anti-inflammatory cytokines, and is closely related to bronchitis. TH1 and TH2 cytokines lung deficiency chronic bronchitis incidence has not been reported. In this study, one animal models syndromes, chronic bronchitis, lung deficiency rat model to improve smoked replication by detecting the content of IL-4 and IFN-γ in serum and lung tissue, and Buqihuoxue mechanism for the law, composed Buqihuoxue party intervention, explore its therapeutic effect, which further reveals evidence of chronic bronchitis, lung deficiency climate essence, and provide experimental basis for clinical prevention and disease card. Methods: 50 Sprague Dawley (SD) rats (male and female) were placed in the experimental environment adaptability fed for one week, were randomly divided into five groups (n = 10): (1) normal control group (normal group, control group); ② CB model group (model group, model group); ③ lungs soup group (the positive control group, prednisone group); the ④ Buqihuoxue side low dose group (low-dose group, low-dose group); ⑤ complement qi and blood circulation the party high dose group (high dose group and high-dose group). According to the literature method using a modified smoked copy chronic bronchitis, lung deficiency syndrome animal model. Modeling groups of rats were placed in a special 1m × 1m × 1m smoke chamber with wood shavings, sawdust, tobacco each 50g ignited fumes, 2 times a day, every 30 minutes for 40 days. The normal control group placed in a normal non-smoking environment rearing. 5 were at the end of the modeling two days after intragastric administration of saline or drugs for 21 days. To the normal group and the model group rats were fed with normal saline (10ml/kg) once a day orally; orally to the low-dose group and high dose groups the Buqihuoxue prescription medicine decoction (12.8g/kg 25.6/kg, equivalent to of clinical adult dosage 7-fold and 14-fold), daily administration; lungs soup Chinese medicinal herbs (8.36g/kg to the lungs soup rats orally equivalent clinical The adult dosage 7 times), once a day, orally. Preparation of the samples tested were: 5 groups were intragastrically 21 days, the animals in each group after the end of treatment, 10% chloral hydrate 0.35ml/100g anesthesia, abdominal aortic blood 3500r/min centrifugal 5 min. Isolated from serum, -20 ℃ cryopreservation to be tested. Take the middle of the right side of the lung tissue 1 cm × 1cm x 1cm size, with 4% paraformaldehyde, embedded in paraffin, sliced. Preparation of the lung homogenates: lung tissue with an ice saline rinse, the filter paper after moisture, said taken wet weight 0.3g added precooled saline, rapid grinding in an ice bath, made of 10% lung homogenate . 4 ℃ 6000r/min centrifugal 5 min extract supernatant, -20 ℃ cryopreservation to be seized. HE staining and SP immunohistochemical staining, lung tissue morphology as well as IL-4 and IFN-γ expression was observed under an optical microscope. Meanwhile, detected by radioimmunoassay in serum and lung tissue of IL-4 and IFN-γ were changes. Results: a general state observed normal rats eating normal, responsive, and body hair smooth and shiny, the two will be normal, and no death phenomenon; model rats there are varying degrees of cough, dyspnea, accompanied by fatigue, curled up, lack of exercise, sluggish, listlessness, hair dry body lean performance;, low and high dose groups, especially with the prolonged treatment time, the spirit of becoming better, breathing stabilized hair increasingly Finishing, increased activity, increased food intake; the lungs soup rats the state also increasingly improved. Bronchus, lung tissue morphology observed 2.1 naked eye observation of normal rats lungs no abnormal appearance lobe outline a clear, smooth, soft surface, pink, good elasticity; model rats lungs was expansive, gray and black spots, volume increases lung surface slightly uneven surface dull, the edge a little spotting; dark red part of the color of the high and low dose group and lungs soup bronchial and lung tissue, occasionally spotting, compared with the model group, a significant difference. 2.2 optical microscope the reference Szapiel of methods to assess bronchial HE staining, the degree of lung tissue inflammatory. Normal group, the integrity of the mucosal epithelium of the bronchioles, cells arranged in neat rows of the bronchioles and alveolar pathomorphology (Fig.1, 2). Model group visible the bronchiolar lumen increased significantly thickened wall, some of the epithelial cell necrosis, associated with chronic inflammatory cells and a small amount of neutrophil infiltration, small venous congestion (Fig. 3, 4). Medication intervention, each treatment group rats bronchitis significantly better than the untreated group. (Fig.5-10). 2.3 immunohistochemistry morphology and semi-quantitative results: IL-4 in the cytoplasm of epithelial cells of the bronchioles of normal rats showed a strong positive expression model rats bronchiolar mucosal epithelial cells in IL-4 was weaker The positive expression (Fig.12). Positive signal showed a brownish-yellow. IL-4 in the cytoplasm of epithelial cells of the bronchioles of the the lungs soup group with the low-dose group showed a weak positive expression (Fig.13, 14); IL-4 expression of the high-dose group showed a strong positive signal. Similar to the normal group, the positive reactions were mainly seen in the bronchiolar mucosal epithelial cytoplasm (Fig.15). The semi-quantitative results show: compared with the normal group, the model of lung tissue in IL-4 significantly reduced (P lt; 0.01); compared with the model group, IL-4 was significantly higher in the lung tissue of rats in the three treatment group ( P lt; 0.01 or P lt; 0.05); comparison between the three treatment groups, the IL-4 levels in the lung tissue of rats in the high dose group and low dose group compared the the lungs decoction group was significantly higher (P lt; 0.01); high doses group compared with the low-dose group was not statistically significant (P gt; 0.05). Bronchiolar mucosal epithelial cells of normal rats showed weak positive expression of IFN-γ (Fig.16); the model rats bronchioles mucosal epithelial cells was a strong positive expression of IFN-γ (Fig .17). Positive signal showed a brownish-yellow. IFN-γ in the cytoplasm of epithelial cells of the bronchioles of the the lungs soup group with the low-dose group showed a strong positive expression (Fig.18, 19); IFN-γ expression of the high-dose group showed a weak positive signal. Similar to the normal group, the positive reactions were mainly seen in the bronchiolar mucosal epithelial cytoplasm (Fig.20). The semi-quantitative results show: compared with the normal group, and IFN-γ in the lung tissue of rats of model group was significantly higher (P lt; 0.01); compared with the model group, IFN-γ was significantly reduced in the lung tissue of three treated rats ( P lt; 0.01 or P lt; 0.05); comparison between the three treatment groups, IFN-γ in the lower dose group and lungs soup group rat lung tissue in the high dose group was significantly higher (P lt; 0.01); low-dose group and lungs soup group showed no statistical significance (P gt; 0.05). (See Table1) 3 groups in serum and lung tissue cytokines IL-4 and IFN-γ content and TH1 and TH2 ratio change 3.1 serum and lung tissue cytokine IL-4 content the normal group, the model rats serum IL-4 were significantly lower (P lt; 0.01); compared with the model group, the three treatment groups serum IL-4 levels were significantly higher (P lt; 0.01) ; comparison between the three treatment groups, the high-dose group, serum IL-4 levels were higher than the the lungs soup group and low dose group (P lt; 0.01 or P lt; 0.05); in the low-dose group, serum IL-4 levels and complement lung soup group was not statistically significant (P gt; 0.05). Compared with the normal group, the IL-4 levels in rat lung tissue in the model group decreased significantly (P lt; 0.01); compared with the model group, IL-4 levels were significantly increased in the lung tissue of three treated rats (P lt ; 0.01); comparison between the three treatment groups, IL-4 levels in the lung tissue of rats of the high dose group and low dose group compared with the lungs soup group was significantly higher (P lt; 0.01 or P lt; 0.05);, high dose group The low dose group was not statistically significant (P gt; 0.05). See Table2, 3.2 groups in serum and lung tissue cytokine IFN-γ were compared with normal group, model group, serum IFN-γ were significantly higher (P lt; 0.01); compared with model group IFN-γ in the serum of three treatment groups were significantly lower (P lt; 0.01); comparison between the three treatment groups, the high dose group and low dose group, serum IFN-γ were significantly lower than the lungs decoction group (P lt ; 0.01 or P lt; 0.05); high-dose group, serum level of IFN-γ in the lower dose group was significantly lower (P lt; 0.01). Compared with the normal group, the model of lung tissue in IFN-γ were significantly higher (P lt; 0.01); compared with the model group, IFN-γ in the lung tissue of the three treatment groups were significantly lower (P lt; 0.01); comparison between the three treatment groups, IFN-γ in the lung tissue of rats in the high dose group and low dose group was significantly lower than the lungs soup group (P lt; 0.01); IFN-γ in the lung tissue of the high-dose group and low dose group not statistically significant (P gt; 0.05). (See Table3) 3.3 serum and lung tissue of TH1 and TH2 ratio changes compared with normal group, model group, serum TH1/TH2 significantly higher (P lt; 0.01); compared with the model group, three-treatment serum TH1/TH2 significantly lower (P lt; 0.01); comparison between the three treatment group was significantly lower than in the high-dose group and the low amount of agent serum TH1/TH2 lungs soup group (P lt ; 0.01); in the high dose group and low dose group, serum TH1/TH2 no significant statistical significance (P gt; 0.05). Compared with the normal group, the model of lung tissue in TH1/TH2 significantly higher (P lt; 0.01); compared with the model group, the lung tissue of rats in the three treatment group TH1/TH2 significantly lower (P lt; 0.01 or P lt; 0.05); comparison between the three treatment groups, the high dose group rat lung tissue TH1/TH2 significantly lower than the the lungs soup group and low dose group (P lt; 0.01); low-dose group and lungs soup group lung tissue of mice TH1/TH2 not statistically significant (P gt; 0.05). (See Table4) Conclusion: chronic bronchitis, lung deficiency state, blood and lung tissue can antagonize the release of inflammatory mediators and reduce airway inflammation cytokines IL-4 levels significantly lower, as well as promote the inflammatory response of cells factor IFN-γ were significantly increased, jointly mediated by non-specific inflammatory response of chronic bronchitis, lung deficiency. Such changes, it is one of the possible pathogenesis of chronic bronchitis, lung deficiency. 2 Buqihuoxue side by elevated serum levels of IL-4 levels in lung tissue, lower serum content of IFN-γ in the lung tissue and TH1/TH2 ratio, in order to weaken or suppress non-specific chronic bronchitis, lung deficiency heterosexual inflammatory response, and prevention and control of chronic bronchitis, lung deficiency.

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