| Research Purpose:On the basis of good efficacy in the clinical application of the strengths of treating antiasthmatic decoction to prevent chronic obstructive pulmonary disease with pulmonary fibrosis, do further study about influence of this medicament on chronic obstructive pulmonary disease with pulmonary fibrosis in animal models of extracellular matrix procollagen type III (PC III), and transparent quality acid (HA) and laminin (LN) content, explore the mechanism action of treating antiasthmatic decoction to treatment of pulmonary fibrosis, and provide a theoretical basis for clinical application.Research Approach:1. Animals and grouping:Total 60 rats all with weight 180-200g—both males and females are 30. Randomly divide them into normal control group, model group, western medicine group, treating antiasthmatic decoction low-does group, high-dose group. There are 12 rats in each group. 2. Animal modeling:The 12 rats are placed in a special fume box (60×55×70cm3), each nine cigarettes phased, one end fixed on the smoke box top and bottom lighting, after complete combustion change another 12 rats, daily 48 rats, weekly 7d, and act 4 weeks continually. In the 5th week, injecting of anesthesia for all rats except in normal group at 10% chloral hydrate (0.36ml/100g) intraperitoneal, shearing conventional iodophor disinfection after the anterior, Line neck incision, blunt separation of the trachea exposed, a one-time injection of bleomycin-side saline solution 0.2ml (bleomycin 4mg/kg body weight) via tracheal cartilage ring gap to the heart puncture. Then let animals upright immediately and rotate mouse plate so that the liquid uniformly distributed in the lungs, skin suture, and routine disinfection. After the animals regained consciousness,put them in cages to breed them conventionally. After modeling from the second day starting medication, the normal group and model group are given normal saline 1ml/150g gavage,2times/day, the western medicine group are given intraperitoneal injection of dexamethasone 0.3mg/100g,1 times/day, treating antiasthmatic decoction low-does group, high-dose group are given free fried granules 4ml, 1ml gavage,2 times/day. All above groups are administered for 28 days.3. Detection methods and observation standard:①generally observe activity, the sensitivity to external reactions, shiny fur, food, water, weight, and deaths of rats.②HA, LN, PCⅢdetected,in the 8th week,after intraperitoneal anesthesia by a 10% chloral hydrate 0.36ml/100g, extract blood strictly according to kit instructions using enzyme-linked immunosorbent assay in rat plasma HA, LN, the level of PCⅢ.③Pathological morphology, kill rats in batches, promptly remove the right middle lobe and fix in 10% neutral formalin. Gradient ethanol dehydration, xylene and transparent specimens, dipping wax, embedding, slicing, and treated by HE, Masson staining to observe the extent of alveolitis and pulmonary fibrosis and count bronchial mucosa fibroblasts, lymphocytes and macrophages under thress high magnifications.4. All results using mean±standard deviation (x±s) to denote, using SPSS10.0 software mean one-way ANOVA to analyze. P<0.05 is considered to be with statistically significant, P<0.01 means significant differences.Research Result:1. The normal control group:smooth fur, able-bodied, diet, activity, body in normal growth. The model control group:dry sparse fur, lack of energy, the reaction retardation, eating, and activities reduced. Wheezing symptoms after strenuous exercise, weight increasing slightly compared with pre-made model, individual ones losing weight. Western Group: adequate at the beginning, dry fur in later period, activities and diet reducing. Treating antiasthmatic decoction group:adequate fur, strong body, normal activity and diet, and the weight increased significantly.2. Rats in normal group:normal structure of lungs, unobvious infiltration around the bronchial inflammatory cell, no thickening of vessel wall muscle, no edema, inflammation, fibrosism and no clear alveolar exudates in alveolar septum. For the model group:acute exudative inflammation, with serous exudation of material full of alveolar septum, the part of the alveolar structural damaged, alveolar fusion, fibroblasts, lymphocytes and collagen fibers gathering in large numbers. Traditional Chinese medicine, the large and low-dose groups were with the same pathologic process as the model group, but the degree and extent of fibrosis were lower than the model group (see the attached 1 to 5).3. In the 8th week PCⅢ, HA and LN level in peripheral blood of model group are higher than the same perioa in the normal group, western medicine and Chinese medicine large and small dose group, all P<0.01. In the 8th week PCⅢ, HA and LN level in peripheral blood of western medicine, Chinese medicine large and small dose group is higher than the normal group in the same period, P<0.05. Western medicine group is no difference from Chinese medicine large and small dose group, P>0.05.4. Counts of fibroblasts, lymphocytes and macrophages in bronchial mucosa of the model group was higher than the normal group, P<0.01. Counts of fibroblasts, lymphocytes and macrophages in bronchial mucosa of the western medicine was lower than the model group, P<0.05. Western medicine group is no difference from Chinese medicine large and small dose group, P>0.05. Consistent with chronic airway inflammation in COPD, and fiber cells in the PF process proliferated into the pathological features.Conclusion:Treating antiasthmatic decoction can effectively ease pulmonary fibrosis of rats. One of the possible role of its mechanisms is to reduce the content of UA, PCⅢ, LN in extracellular matrix of pulmonary fibrosis rats.
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