| Asthma is a chronic inflammatory disorder of the airways. Its essence is the abnormal immune response of a disease caused by a variety of cells, inflammatory mediators and cytokines network participation. As a serious harm to the health of the people of all ages and complexity of refractory disease, its morbidity is increasing year by year and it has become one of the focuses of current research.Chinese medicine for the treatment of asthma has a long history and its own characteristics. Making up for the western medicine in treating bronchial asthma with higher prices,bigger poisonous side effect and the need for long-term use,it has been recognized by people in medical practice. The external therapy of traditional Chinese medicine has its unique advantages. It can achieve the effect which internal can not and help to reduce the period of onset of asthma symptoms,especially play an important role in the prevention and the remission stage of treatment of asthma.As a new type of babu agent for external use, it has been widely used with virtue of the unique advantages of its own. MJPC used in this paper is made of white mustard seed, rhizoma corydalis, ephedra, bitter almond, ginger juice. As a new preparation for treating asthma, MJPC is ordered for the asthma exacerbation and remission. Therefore, this paper will be made from skin safety research, effect on bronchus spasmolysis and anti airway inflammation, study on the mechanism. Through the experimental research of four aspects, we can evaluate the safety and efficacy of MJPC and make the mechanism of the effect clear.Objective1 Establish different species of animal model of skin lesions, in order to evaluate the safety of MLPC.2Observe the effect on isolated guinea pig trachea smooth muscle, in order to explore the mechanism of MJPC relieving bronchial spasm.3 Establish BN rat model of asthma,in order to observe the effect of MJPC on the animal model of asthma.4 Through the further study of the change of TSLP, explore the mechanism of action of MJPC for the treatment of asthma.Methods:1 Observe animal acute toxicity, skin irritation and allergy medication of MJPC by the acute toxicity test and skin irritation in rabbits, guinea pigs in experimental allergic skin test.2Prepare isolated guinea pig tracheal spiral strips,in order to respectively observe the antagonistic effect of MJPC on histamine and acetylcholine induced contraction of guinea pig tracheal smooth muscle.3 Prepare BN sensitive asthma model of rats induced by OVA in order to observe the effect of MJPC in peripheral blood and bronchoalveolar lavage fluid white blood cell count, lung function and lung tissue injury.4By the preparation of BN sensitive asthma model of rats induced by OVA, immunohistochemistry and radioimmunoassay method was used to observe the effect of MJPC of TSLP, BALF in rat serum, thymus, skin and lung tissues.Results:1 Safety of MJPC skin medication:MJPC to rabbit skin and damaged skin did not cause acute toxicity; mild irritation to rabbit skin and damaged skin, but after administration of 48h, the stimulation disappeared; sensitization effect on the skin of guinea pig.2The MJPC on isolated guinea pig trachea smooth muscle:MJPC composition of acetylcholine chloride and phosphoric acid histamine induced contraction of tracheal strips of guinea pig in vitro have obvious antagonism, and it had obvious dose effect relationship3 Antiasthmatic effect of MJPC:MJPC treatment group and positive drug dexamethasone (Dex) could prolong the OVA sensitized rats in the incubation period of asthma. Compared with the model group, MJPC treatment group and Dex positive group were significantly different (P<0.05).4 Pulmonary function test results:compared with the blank group, the model group has significant differences of 3.125mg/ml,6.25mg/ml and 12.5mg/ml at the concentration of Mch (P<0.01). MJPC treatment group and positive drug dexamethasone (Dex) can reduce the airway responsiveness to Mch. Compared with the model group, MJPC treatment group with a very significant difference in Mch concentration of 3.125mg/ml and 6.25mg/ml (P<0.01), at the concentration of Mch was 12.5mg/ml with a significant difference (P<0.05).5 EOS in blood count results:in the model group, the peripheral blood eosinophil count increased, compared with normal group had significant difference (P<0.01). MJPC treatment group and positive drug dexamethasone (Dex) can reduce the asthma induced by OVA in rat peripheral blood eosinophil count. Compared with the model group, MJPC treatment group and Dex positive group had significant difference (P<0.01).6 The counting result WBC of BALF:model group, eosinophil, neutrophil and lymphocyte counts were significantly increased, compared with normal group had significant difference (P<0.01). MJPC treatment group and Dex positive group could reduce the eosinophil, neutrophil and lymphocyte counts in BALF rats, compared with the model group, with significant difference treatment of mustard and MJPC group (P<0.05). In model group, the number of monocytes decreased, there was significant difference compared with the normal group (P<0.05).MJPC treatment group and Dex positive group significantly increased the number of mononuclear cells in BALF rats, compared with the model group, with significant difference treatment of mustard and Ma Babu group (P<0.05).7The pathological section of lung tissue HE staining results:blank group, no inflammatory cell infiltration, airway epithelial thickening, alveolar wall structure. The model group and the tube wall of airway mucosa showed obvious inflammatory cell infiltration, airway wall thickening and shedding of epithelial cells. Compared with the model group,MJPC treatment group and Dex positive group than in mild airway inflammation, airway wall with a small amount of inflammatory cell infiltration in lung tissue, there was no obvious pathological injury.8 The results of immunohistochemistry:Lung tissue:the integral optical density of TSLP in the lung tissue of model group increased, compared with the control group, with significant difference (P<0.01). MJPC treatment group and Dex positive group could reduce the integral optical density of OVA TSLP in lung tissue of rats in the value sensitivity. Compared with the model group, with significant difference of MJPC treatment group and Dex positive group (P<0.05).Skin:skin integral optical density of TSLP in model group increased, compared with the control group, with significant difference (P<0.01). MJPC treatment group and Dex positive group could reduce the integral optical density of OVA induced by TSLP in rat skin resistance value. Compared with the model group, with significant difference of MJPC treatment group and Dex positive group (P<0.05).9 The Elisa content of serum TSLP, BALF, lung tissue, skin and thymus in test results:Serum:serum TSLP levels in the model group, compared with the control group with significant difference (P<0.05). MJPC treatment group and Dex positive group were able to reduce the content of TSLP level in serum of rats induced by OVA sensitive. Compared with the model group, with a significant difference between Dex positive group (P<0.01), significant differences in treatment of mustard and MJPC group (P<0.05).BALF:the elevated level of TSLP BALF in the model group, compared with the control group with a very significant difference (P<0.01). MJPC treatment group and Dex positive group can reduce the TSLP content in BALF of rats induced by OVA sensitive. Compared with the model group, with a significant difference between Dex positive group (P<0.01), significant differences in treatment of mustard and MJPC group (P<0.05).Lung tissue:the increase of TSLP level in the lung tissue of model group, compared with the control group with a very significant difference (P<0.01). MJPC treatment group and Dex positive group could reduce the content of TSLP in lung tissue of sensitivity level of rats induced by OVA. Compared with the model group, with significant difference of MJPC treatment group and Dex positive group (P<0.05).Skin:skin increased the level of TSLP in the model group, compared with the control group with significant difference (P<0.05). MJPC treatment group and Dex positive group can reduce the TSLP level in rat skin allergy caused by OVA. Compared with the model group, with significant difference of MJPC treatment group and Dex positive group (P<0.05).Thymus:increased the level of TSLP in thymus in the model group and blank group, but no significant difference (P>0.05). MJPC treatment group and Dex positive group significantly increased OVA TSLP levels in rat thymus sensitivity. Compared with the model group, with significant difference of MJPC treatment group and Dex positive group (P<0.01).Conclusion:1 The MJPC is non-toxic with clinical medication safety.2 The MJPC has good antiasthmatic effect. In vitro experiments showed that MJPC drug components caused by histamine phosphate and acetylcholine chloride guinea pig tracheal smooth muscle contraction has antagonistic effect significantly.3 The MJPC on BN rat asthma animal model can play a therapeutic role. Pulmonary function test results show that the MJPC for Mch induced airway smooth muscle spasm had better antagonistic effects; it can significantly reduce the variety of EOS and neutrophils number; HE staining showed that the lung pathological injury can be reduced in asthma animal model. 4Treatment of MJPC on BN rat asthma model is realized by adjusting the level of TSLP in different tissues of. Lung tissue and skin immunohistochemical detection of TSLP, BALF and serum, skin and lung tissue homogenate TSLP test results show that, MJPC treatment group has low expression of TSLP and TSLP expression is higher in model group, MJPC can be regulated by different tissue TSLP level and therapeutic effect for asthma animal play.It is visible that MJPC on BN rat animal model of asthma can play a good therapeutic effect, and by regulating the level of TSLP in different tissues of the animal it plays a role in the treatment of asthma. The effect on the treatment of asthma and other regulatory mechanisms need to be further confirmed in clinical.
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