| Objective:Changfeifangxiao Decoction is an experiential effective prescription in the treatment of asthma for many years.It had been applied to clinic,and the effect is significant.It's immune function has an initial exploration.This research studies on Bronchus,lung tissue morphology, ieosinophils (EOS), IgE and eosinophil chemotactic protein (Eotaxin), intercellular adhesion molecular-1(ICAM-1) expression levels in their blood and BALF through in vivo experiment. Explore Changfeifangxiao Decoction's mechanism of action in the treatment of asthma further from the cellular and molecular level,and lay the foundation for development of an effective clinical prevention and treatment of asthma Chinese herbal compound preparation.Methods:(1)Preparation of Traditional Chinese MedicineExtract volatile oil from Herbal Ephedrae,Radish Seed,Fructus Aurantii.Then,dregs of Herbal Ephedrae,Radish Seed,Fructus Aurantii with the remaining prescription drugs and water decoct three times.concentrate into extract after filter.Proportional to join a certain amount of volatile oil before Mixing for experimental animals intragastric administration.(2) Grouping and Modeling6-week-old SD male rats of clean grade 60, weight 150±20 g, were randomly divided into six groups, each group 10 rats.A group is normal group.B,C,D groups are Changfeifangxiao Decoction large, medium and small dose group.E group is a drug (dexamethasone) group. F Group is a patho-model group.Rats hypodermic injection with lml physiological saline containing 1mg OVA and 200mg aluminum hydroxide while intraperitoneal injection with Bordetella pertussis vaccine 6×109 samples were taken for immunostimulants.On the 15th day,Use 2% OVA to nebulize and inhale once a day, once 30 minutes, for 7 days. Nebulize and inhale physiological saline for A group.(3)Dose RegimensGroups begin intragastric administration at the 8th day, Changfeifangxiao Decoction were given the large doses of 33.84 g crude drug/kg/d for B group,the medium dose of 16.92 g crude drug/kg/d for C group,small dose of 8.46 g crude drug/kg/d for D group.E group were given dexamethasone 0.75 mg/kg/d. A and F group were given commensurable physiological saline. Capacity of dosage are 20 ml/kg, for 13 days.(4) SamplingAfter the last of induced asthma 24 hours,rats were anesthetized and carotid artery blood 3ml. Ligate of the left lung,bronchial intubation. In the right lung for bronchoalveolar lavage(2ml,3 times), collect bronchoalveolar lavage fluid (BALF).Use the physiological saline to wash blood vessels,then use the paraformaldehyde perfusion fixation.Take the right middle lung embedded in paraffin and sliced. (5)Detecting①Take bronchoalveolar lavage fluid (BALF) 0.1ml to count on the cell count plate.The rest of the BALF centrifuge with the speed of 1200 r/min in the centrifuger for 10 minutes.Sediment smear on the cell count plate,then count differentially.Liquid supernatant is stored at-20℃.②Take blob of blood to smear,and count differentially to the number of blood cells. Measure serum levels of IgE by radioimmunoassay.③Bronchial lung biopsy use routine HE staining,and observe through the optical mirror. Analyze the situation of airway remodeling by image analysis software.Results:1.Local airway inflammation and the changes of bronchoalveolar lavage fluid,blood in cytology1.1 The pathological section of lung tissue by HE staining showed, Lung tissue eosinophils and inflammatory cell infiltration by small airway remodeling in Changfeifangxiao Decoction large, medium and small dose groups and dexamethasone group are more reduced significantly than patho-model group.1.2 The cell count of bronchoalveolar lavage fluid showed, Changfeifangxiao Decoction large, medium and small dose groups and dexamethasone group can eosinophil counts and the percentage of significantly reduced. There are significantly different from patho-model group (P<0.05,P<0.001).1.3 The cell count of blood showed,Changfeifangxiao Decoction large, medium and small dose groups and dexamethasone group can eosinophil counts and the percentage of significantly reduced. There are significantly different from patho-model group (P<0.01,P<0.001).2.The changes of eosinophil chemotactic protein (Eotaxin), intercellular adhesion molecular-1(ICAM-1)in bronchoalveolar lavage fluid,and IgE in peripheral blood.2.1 Changfeifangxiao Decoction large, medium and small dose groups and dexamethasone group can significantly lower the content of Eotaxin.There are significantly different from patho-model group(P<0.01,P<0.001).2.2 Changfeifangxiao Decoction large, medium and small dose groups and dexamethasone group can significantly lower the content of ICAM-1.There are significantly different from patho-model group(P<0.05,P<0.001).2.3 Changfeifangxiao Decoction large, medium and small dose groups and dexamethasone group can significantly lower the content of IgE.There are significantly different from patho-model group(P<0.001).Conclusion:Changfeifangxiao Decoction achieve the treatment of asthma by reducing the EOS infiltration and the number of EOS,inhibiting Eotaxin and ICAM-1 expression, reducing levels of IgE. |
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