Structural Changes In The Lung Tissues Of COPD With Lung-qi Deficiency Syndrome In Rats And The Intervention Study Of Ailuo Kechuanning Decoction

ObjectiveTo study the effect of Ailuo Kechuanning Decoction on lung function, inflammation, tissue remodeling in rats with lung qi deficiency syndrome model of Chronic Obstructive Pulmonary Disease (COPD). To clarify the therapeutic effect and mechanism of this prescription on COPD, and provide experimental basis for clinical application and development of new drug.MethodsFifty SD rats were randomly divided into 5 groups, each group of 10 animals, including the normal group, model group, Ailuo Kechuanning Decoction of low dose (7.76g·kg-1·d-1) group and middle dose (15.52g·kg-1·d-1) group, high dose (31.04g·kg-1·d-1) group. Preparation for COPD in rats with lung qi deficiency syndrome models by using cigarette smoke and dripping lipopolysaccharide in the trachea. After making the model successfull, treated by intragastric administration observing and recording the general situation of rats in each group, and detectting the lung function [forced vital capacity(FVC), forced expiratory volume in one second (FEV1) and FEV1/FVC]. To detect interleukin-6 (IL-6) and soluble intercellular cell adhesion molecule-1 (sICAM-1) in lung tissue homogenate of rats and bronchoalveolar lavage fluid(BALF) by Enzyme-Linked ImmunoSorbent Assay(ELISA) and observation of rat lung tissue under light microscope. Using immunohistochemistry (immunohistochemistry, IHC) method to detect the expression of transforming growth factorl(TGF-betal,) and it's receptor (TGF-beta RI)protein, Smad3, Smad4,Smad6 and Smad7, matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), type I, type III collagen protein that all relating to the deposition of extracellular matrix(ECM) in the lung tissue. To observe the expression of Smad3, Smad4, Smad6 and Smad7 mRNA by quantitative real time PCR (RT-PCR) method.ResultsCompared with the normal group:the FVC, FEV1 and FEV1/FVC of the model group were significantly decreased(P<0.01), the content of IL6 and sICAM-1 in BALF and homogenate of lung tissue were all significantly increased (T<0.01); the expression of mRNA Smad3 and mRNA Smad4 in model group was increased (P<0.05), the expression of Smad6 mRNA and Smad7 mRNA were significantly decreased (P<0.05), the expression of Smad4 and Smad3 protein was significantly increased (P<0.01), the expression of Smad7 and Smad6 protein was significantly decreased (P<0.01); in model group, the expression of TGF-1 in lung tissue was significantly increased (P<0.01), the expression of TIMP-1 and MMP-9 was significantly increased (P<0.01), the expression of type III and type I collagen in lung tissue was significantly increased (P<0.01).Compared with model group:Ailuo Kechuanning Decoction low, middle and high dose groups of FVC, FEV1 and FEV1/FVC index were increased (P<0.01), lung tissue homogenate and BALF of IL-6 and sICAM-1 levels were lower, and the Ailuo Kechuanning Decoction middle dose group decreased significantly (P< 0.01); Ailuo Kechuanning Decoction high middle and low dose group of Smad3 mRNA and Smad4 mRNA expression was significantly decreased (P< 0.05), the expression of Smad6 mRNA and Smad7 mRNA display increased (P< 0.05), Smad3, Smad4 protein expression was significantly decreased (P<0.01), Smad6, Smad7 protein expression was significantly increased (P<0.01); Ailuo Kechuanning Decoction of low, middle and high dose group TGF-betal and TGF-beta RI protein expression decreased significantly (P<0.01), MMP-9, TIMP-1 expression was significantly decreased (P<0.01), type ?, type ? collagen expression was significantly decreased (P<0.01).Under the light microscope, the model group had sputum embolism, mucosal epithelial degeneration and necrosis.Submucosal gland hyperplasia, hypertrophy, glandular cavity mucus filled with irregular expansion, the wall around the wall and interstitial thickening of the lung, a large number of inflammatory cell infiltration; Interstitial thickening of the peripheral pulmonary alveolar, alveolar interstitial. Compared with the model group, Ailuo Kechuanning Decoction agent group fine bronchial mucosa with different degree of repair, more regular arrangement of epithelial, submucosal gland volume became small number of reduce, inflammatory cells disappeared; Bronchiole alveolus between qualitative thin, locally infiltrating lymphocytes; Ailuo Kechuanning Decoction high dose group and low dose group of deviation to the recovery of the middle dose group.Conclusions1. Ailuo Kechuanning Decoction can significantly improve lung function in rats with lung qi deficiency syndrome COPD.2. Ailuo Kechuanning Decoction can be effective in the treatment of COPD lung qi deficiency syndrome in rats inflammatory reaction and protect lung tissue and alleviate lung deficiency cough, sputum and other symptoms. The mechanism may be related to reducing the secretion of cytokines IL-6 and sICAM-1, inhibiting the inflammatory reaction and preventing the progression of the disease.3. Ailuo Kechuanning Decoction can effectively inhibit bronchial remodeling, improve pulmonary ventilation, has antiasthmatic effects. The mechanism may be by reducing the Smad3 gene expression and enhance the expression of Smad6 and Smad-7 gene, coordination of Smad4 gene expression, inhibit the expression of TGF-1 and its receptor and inhibit the expression of MMP-9 and and coordination to inhibit the expression of TIMP-1 protein, alleviate the bronchioles and pulmonary tissue remodeling.