| [objective]Exploring the influence of tonifying lung decoction on the stable period of rats with COPD trachea remodeling, verifying whether tonifying lung decoction can regulate immune function and inhibit trachea inflammation through TLR2/NF-?B signal transduction pathway of rats with COPD, providing experimental basis for the clinical application of tonifying lung decoction, thus, demonstrating the feasibility and importance of taking "strengthening lung and benefiting kidney" method as the main treatment method of traditional Chinese medicine (TCM) for the stable period of COPD further, understanding the correlation between the syndrome type of TCM——"qi deficiency of lung and kidney" and objective indicators, such as "immunocomprosised" and "trachea inflammation" deeply, deepening the understanding of essence of the stable period of COPD——"qi deficiency of lung and kidney", promoting objectification, standardization and normalization of syndrome types of TCM, showing significance to guide basic research and clinical application of the TCM therapy of COPD.[methods]84 rats were divided into 7 groups:control group, model group, the high dose of tonifying lung decoction of TCM granules group (hereinafter referred to as granules high group), the middle dose of tonifying lung decoction of TCM granules group (hereinafter referred to as granules middle group), the low dose of tonifying lung decoction of TCM granules group (hereinafter referred to as granules low group), the tonifying lung decoction of TCM liquid group (hereinafter referred to as liquid group), positive control western medicine Montelukast group (hereinafter referred to as western medicine group), with 12 in each group randomly. All the rats except for those in control group were injected lipopolysaccharide (LPS) into tracheas at the first day and the 25th day from the beginning of the experiment, didn't accept any processing in the 2-8th days, the 23-24th days, and the 26-28th days for the rats restoring physical strength, were smudged 1 time per day in the 9-22nd days and the 29-36th days,2 times per day in the 37-44 days, and 3 times per day in the 45 to 52 days. The rats in control group were injected the same amount of normal saline (NS) into tracheas at the first day and the 25th day, and didn't have passive smoking. After the success of modeling, gavage administration began from the second day. Control group and model group were given NS (10ml/kg·d), western medicine group was given Montelukast (1.0 mg/kg·d), granules high group, granules middle group and granules low group were given respectively tonifying lung decoction of TCM granules (27.8 g/kg·d?13.9 g/kg· d?7.0 g/kg·d), and liquid group was given tonifying lung decoction of TCM liquid (13.9 g/kg·d), for 32 days in a row. All rats were incided in the femoral artery from the 33rd day, bled to death, took the right lung tissue to observe the pathological change.detect TLR2, TLR4, MyD88, and NF-?B positive expression of protein genes by immunohistochemical SP method, and detect the content of TLR2, MyD88, and NF-?B in the lung tissue by Western Blot method, took the bronchoalveolar lavage fluid (BALF), and detect the level of IL-1 ??IL-6?IL-8?NF-?B?TNF-? inflammatory factors in BALF by ELISA method.[results]1. General situation:the rats in control group recovered well in the wound, were with little scar, good nutritional status, normal mental, eloquent eyes, white and lustrous fur and smooth breathing, were active and responsive, their eating, drinking and defecation were normal, they were without symptoms such as coughing and gasping, etc, without abnormal secretions in the respiratory tract, without wheezing sound; while the rest of the groups appeared gradually during the process of modeling poor nutritional status, decreased activity, depression, dull eyes, slowing to respond, less eating, loose stool, gloomy, vertical and easy to fall off fur, secretions discharged from the corners of the mouth sometimes, and the sound of coughing and wheezing could be heard; each treatment group was improved in the above situation after the treatment, among which the most obvious group was granules high group, followed by granules middle group and western medicine group.2. Pathological examination:the rats in control group were with complete tracheal mucosa ciliated columnar epithelium cells, respiratory tract and alveolar epithelial, regular cilia, a few scattered lymphocytes infiltrating in all levels of trachea and bronchi, uniform alveolar, proper thickness of alveolar interstitial, rarely macrophages in alveolar area, hardly any secretions and inflammatory cells in the bronchial lumen. The rats in model group were with incomplete mucosa epithelial, part stripped tracheal and bronchial ciliated epithelium, significantly hyperplasia and hypertrophy of goblet cells and glands, increased and extended bronchial mucosal fold into the lumen; inflammatory cells infiltration in all levels of bronchial mucosa and submucosa, mainly lymphocytes and macrophages, in addition, plasma cells, and small amounts of neutrophils, thickening of bronchial smooth muscle, mucous plugs and numerous neutrophils in the small bronchial lumen; there were a lot of macrophages which engulfed dust particles (dust macrophages) beside small bronchi and pulmonary artery, part macrophages collapse and destroyed; far-end of terminal bronchiole was narrow, respiratory bronchioles and alveolar ducts were cystic dilatation, alveolar space irregular expand, alveolar wall became thinning, swelling, and ruptured, or fused, so pulmonary bulla formed. Compared with model group, these lesions alleviated in granules high group, granules middle group and liquid group.3. Detecting inflammatory factors by ELISA method:compared with control group, the contents of all of IL-1??IL-6?IL-8?NF-?B and TNF-a in BALF of rats in model group have significant difference. Compared with model group:the content of IL-1? in BALF of rats in granules high group, granules middle group, liquid group and western medicine group has significant difference; the content of IL-6 in BALF of rats in granules high group, western medicine group and granules middle group has significant difference; the content of IL-8 in BALF of rats in granules high group, liquid group and western medicine group has significant difference; the content of NF-?B in BALF of rats in granules high group and western medicine group has significant difference; the content of TNF-a in BALF of rats in granules high group, granules middle group, liquid group and western medicine group has significant difference.4. Detecting protein gene positive expression by immunohistochemical SP method:compared with control group, the expression of all of NF-?B,TLR2, TLR4 and MyD88 has significant differences in model group. Compared with model group:the expression of NF-?B has significant differences in liquid group, western medicine group and granules high group; the expression of TLR2 has significant differences in liquid group, western medicine group, granules high group and granules middle group; the expression of TLR4 has significant differences in liquid group, western medicine group, granules high group and granules middle group; the expression of MyD88 has significant differences in liquid group, granules high group and granules middle group.5. Detecting the contents of protein by Western Blot method:compared with control group, the contents of all of MyD88?NF-?B?TLR2 in model group have significant difference. Compared with model group:the content of MyD88 in western medicine group, liquid group, granules high group and granules middle group has significant difference; the content of NF- ?B in western medicine group, liquid group, granules high group and granules middle group has significant difference; the content of TLR2 in western medicine group, granules high group and granules middle group has significant difference.[conclusions]1. Both tonifying lung decoction of TCM granules and liquid can inhibit activity of TLR2 and TLR4, which are the two kinds of toll-like receptors in lung tissue of COPD rats at the same time, regulate downstream NF-?B signal transduction pathway by classic way of MyD88 dependence, reduce the release of inflammatory factors such as IL-1??IL-6?IL-8? NF-?B?TNF-?, etc, thus, improve trachea remodeling, but which is mainly through TLR2 pathway and probably with joint effect of TLR4 pathway.2. Tonifying lung decoction has the dose dependency to improve trachea remodeling of COPD through regulating TLR2/NF-?B signal transduction pathway. |
PDF Full Text Request