| ObjectiveThis study attempts to animal experiments, in the "The lung and the large intestine being interior-exteriorly related" under the guidance of TCM(Traditional Chinese Medicine) theory, the establishment of similar rats with clinical prototype, "Fei Chang disease" (asthma together constipation) model, as the "The lung and the large intestine being interior-exteriorly related" Theoretical basis for experimental study, animal model to compare the observed "Fei Chang disease" Characterization and biological indicators of change, and the corresponding changes in the relationship between the evaluation system to establish the model.MethodsSPF-class selection of 40 healthy male Wistar rats were randomly divided into 4 groups:control group, "lung disease" (asthma) model group, "bowel disease" (constipation) model group, "Fei Chang disease" (asthma together constipation) model group, 10rats in each group. "lung disease" (asthma) Preparation of model group, the OVA 1mg and aluminum hydroxide 200 mg in normal saline 1ml of fresh gel preparation of sensitization, day 1 and day 8 in rats with bilateral chest, groin, a total of 4 points in the sensitization of the subcutaneous injection of 0.15ml,0.4ml intraperitoneal injection of a total of lml sensitized. The 15th day the rats placed in plexiglass boxes, ultrasonic nebulized 1% OVA-induced asthma, day 1, every 30 min. atomization 1w, a total of 21 days. "Bowel disease" (constipation) Preparation of model group, fed Compound Diphenoxylate tablets (DC), dose of 10mg/kg, day 1. "Fei Chang disease " (asthma together constipation) model group, the preparation, oral administration of Compound Diphenoxylate (a dose of 10mg/kg), day 1; same time, the OVA lmg and 200mg of aluminum hydroxide dissolved lml of prepared saline solution into the gel sensitization, day 1 and day 8 in rats with bilateral chest, groin, a total of 4 points in the sensitization of the subcutaneous injection of 0.15ml. 0.4mlintraperitoneal injection were caused by a total of lml Min. The 15th day the rats placed in plexiglass boxes, ultrasonic nebulized 1% OVA-induced asthma, day 1, every 30 min. atomization 1w, a total of 21 days. Control group was given distilled water and ultrasonic atomizing inhalation with normal saline. Preparation of "Fei Chang disease " model group, the detection of rat pulmonary, gastrointestinal, lung and other physiological and pathological indicators of intestinal flora, light and electron microscopy tissue HE staining and ultra structural pathological changes of circumstances.Results1. Degree view from asthma, "Fei Chang disease" group than the control group the extent of asthma, "lung disease " group is serious and there was significant difference (P< 0.01).2. Rat stool observation:with the control group, "lung disease " group, "Fei Chang disease "group rats were significantly decreased fecal wet weight, fecal particles decreased, there was significant difference (P< 0.01); and "bowel disease" group. "Fei Chang disease " group promoting the value of the intestinal, fecal wet and dry weight were reduced, reducing the number of fecal particles, there are differences (P< 0.05).3. Blood test:comparison with the control group, "Fei Chang disease " group of leukocytes, neutrophil counts were significantly increased, lymphocyte count and intermediate cells increased, there was significant difference (P< 0.01); and "bowel disease" group, "Fei Chang disease " group of leukocytes, significantly higher neutrophil count, lymphocyte cell count and the number of intermediate cell count increased, there was significant difference (P< 0.01):and "lung disease " group, "Fei Chang disease " group of leukocytes, lymphocytes no difference (P> 0.05). neutrophil count increased and the intermediate cells, there are differences (P< 0.05).4. Pulmonary function:compared with the control group, "Fei Chang disease group of the respiratory rate increased, decreased minute ventilation, tidal volume decreased (P< 0.01);and "bowel disease" group phase Ratio, "Fei Chang disease' group increased respiratory rate, minute ventilation decreased, tidal volume decreased (P<0.01); and "lung disease " group, "Fei Chang disease " rats Respiratory rate increased, decreased minute ventilation, tidal volume decreased (P< 0.05).5. Gastrointestinal function tests:comparison with the control group, "Fei Chang disease " rate of intestinal propulsion in rats reduced gastric residual rate was significantly higher (P< 0.01); and "bowel disease" group, "Fei Chang disease " rate of intestinal propulsion in rats reduced gastric residual rate was significantly higher (P < 0.05); and "lung disease " group, "Fei Chang disease " rate of intestinal propulsion in rats reduced, gastric residual rate increased (P< 0.01).6. Light and electron microscopy in the lung and gastrointestinal tissue pathological changes:(1) Light microscopy:"Fei Chang disease", the normal structure of lung tissue is damaged, disordered markings, lung cell degeneration and necrosis, there swelling, local congestion, edema, mild mucosal erosion, most of the following segmental bronchi alveolar and more inflammatory cell infiltration, interstitial pulmonary fibrosis in a large organization there are a large number of monocytes, lymphocytes, eosinophil's such as cell infiltration. Performance of gastrointestinal tissues; intestinal mucosa due to complete due to the rules arranged glands, mucous glands damage, mucosal and submucosal inflammatory cell infiltration. (2) Observation:"Fei Chang disease" lung cells in rats appeared typical structure of cell damage, the gap widened adjacent cells, the cell was atrophy, dilated large endoplasmic reticulum vesicles, mitochondrial swelling, morphology irregular; gastrointestinal cells showed the typical structure of cell damage, mitochondrial swelling, irregular shape, gastrointestinal cell surface microvilli disappearance or reduction, and more large and dense fibrous septa, straight edge.7. Pulmonary intestinal flora detection:comparison with control group, "Fei Chang disease" group increased the number of lung aerobic bacteria and fungi, anaerobic bacteria did not change significantly (P< 0.01); Intestinal aerobic Elevated E. coli bacteria, anaerobic bacteria, fungi, Bacteroides and Bifid bacterium decreased (P<0.01). And "bowel disease" group, "Fei Chang disease " group increased the number of lung aerobic bacteria and E. coli (P< 0.01), anaerobic bacteria did not change significantly (P< 0.05):intestinal aerobic Elevated E. coli bacteria, anaerobic bacteria, fungi, Bacteroides and Bifid bacterium decreased (P< 0.05). And "lung disease " group, "Fei Chang disease " group increased the number of lung aerobic bacteria and fungi (P< 0.01), anaerobic bacteria did not change significantly (P< 0.05); intestinal aerobic and Fungi increased (P<0.01), anaerobic bacteria, E. coli and Bifid bacterium Bacteroides no significant change (P< 0.01).Conclusions1. From the general state of rats, changes in living habits, and related physiology, pathology, biochemical test observations, "Fei Chang disease " model of the degree of asthma and constipation in rats, pathological changes and morphological changes in lung intestinal flora Than those of pure "lung disease" group and simply "bowel disease "group seriously, initially confirmed that the "Fei Chang disease" composite model of success.2. The establishment of the "Fei Chang disease" model rats, both "lung disease "(asthma) together, "bowel disease "(constipation) is characterized, for further in-depth study of "The lung and the large intestine being interior-exteriorly related" laid the mechanism Basis.3. From the macro and micro indicators are characterized by an objective assessment, "Fei Chang disease" status and the associated pathophysiological changes, some reveal a "lung" and "bowel" the possible interactions, and the impact mechanism for" lung Intestinal disease together, "Establishment and evaluation of a rat model provides a basis.4. "Fei Chang disease" rat model for the study of other complex animal model and provides a reference. |
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