The Research Of Mechanism On Cough Of Airway Neurogenic Inflammation In Rats After Infection For Xuanfeizhisou

Objective:Through the study of the effect of Xuanfeizhisou and the related mechanism, to provide experimental basis for the clinical application of the scientific party; Based on the study on the method of establishing the animal model of cough after infection, in order to provide a stronger operational for making the animal model; And on this basis, through animal experiments to observe the regulation of airway neurogenic inflammation in PIC rats for Xuanfeizhisou, to explore the specific mechanism, conducive to the development of the theory and practice of Chinese medicine treatment of this disease.Methods:(1) Using the method of ammonia water induced cough,phenolsulfonphthalein excretion test to observe the effects of antitussive and expectorant of Xuanfeizhisou with mice; Methods is used to replicate the model of chronic bronchitis, to observe the influence on the morphology of lung tissue of mice for the side.(2) The rats are divided into control group and cough after infection model of A, B, C, D, group, small dose of lipopolysaccharide(LPS) induces by simulated leather method of gram-negative infections created PIC animal model. Observing the behavior changes of rats and cough properties, detecting the total number of BALF cells and the counting and classification, determining the cough response; and observing the pathological changes of lung tissue by HE staining.(3) The SD rats are randomly divided into6groups, namely control group, PIC model group, positive control group and Xuanfeizhisou of low, middle and high dose group. The rats are killed after Intragastric administration of15d. Detecting the total number of BALF cells and the counting and classification;Observing the pathological changes of lung tissue by HE staining;ELISA method is used to detect BALF speed of bradykinin (SP, NKA, NKB) and calcitonin gene related peptide (CGRP) content; and detecting the expression of protein of SP, SP receptor (NK-1) and NKA in lung tissues by immunohistochemical method.Results:(1) With the increase of dosage of Xuanfeizhisou, the cough incubation period can be prolonged in mice. High dose is more significantly (P<0.05); Inhibition of mouse cough. High, in dose effect is more obviously (P<0.01), the low dose is no statistically significant difference, and a certain dose-effect relationship; The phenol red excretion quantity of mouse tracheal is significantly increased (P<0.01), also shows a dose-effect relationship; Reduction the change of the model of mouse lung tissue pathologic.(2) The study on the method of establishing the animal model of cough after infection shows:â‘ Model of group A and group B rats coughing frequently, and the movements of cough are more typical, and model group B is more obvious.â‘¡Determination of cough reactive:each model group compared with the control group5min cough have increased significantly (P <0.001), but the data analysis shows that group A, group B more significant difference.â‘¢The model of A and B group BALF in various inflammatory cell ratio compared with blank control group are significantly different (P<0.05),in which B group is more significantly (P<0.01).â‘£The pathological of model group A and group B rats shows airway inflammation, it is more obvious in the model group B especially, while there is no significant pulmonary interstitial inflammation and alveolar fusion.(3) The study on the influence of airway neurogenic inflammation in rats of cough after infection for Xuanfeizhisou shows:â‘ The treatment group compared with the model group, the ratio of various inflammatory cells in BALF decreases, with Xuanfeizhisou in agent group data change is the most significant (P<0.001), followed by low dose group, high dose group; While the Robitussin group compared with the model group, the difference is not statistically significant.â‘¡Detecting the content of BALF supernatant medium kinin SP, NKA, NKB and CGRP can be found:Compared with the normal control group, the model group BALF index are significantly increased (P<0.001); the content of BALF index in each treatment group are lower than model group, with Xuanfeizhisou in agent group is decreased most significantly (P<0.001), followed by low dose group; Xuanfeizhisou high and Robitussin group compare with model group, no significant difference.â‘¢Pathological results shows:The airway inflammation and epithelial injury of rats in model group are more significant, while no obvious pathological changes of lung parenchyma; Each treatment group is compared to the model group, the airway inflammation and epithelial injury are improved, and the airway of rats in Xuanfeizhisou middle dose group are improved in the most significant, close to the airway in normal rats, followed by Xuanfeizhisou low dose group; while the Xuanfeizhisou high dose group and Robitussin rats shows similar airway.â‘£The immunohistochemical staining shows:Compared with the blank control group, model group, Robitussin group and Xuanfeizhisou high dose group rats lung tissue show deep yellow secretions positive reaction, while it is shallow in the lung tissue of rats in Xuanfeizhisou middle and low dose groups. The results of immunohistochemistry semiquantitation shows:Compared with the normal control group, the lung tissue of rats in model group SP, NK-1and NKA protein expression are significantly enhanced (P<0.001). Compared with the model group, the positive expression of indexes in Xuanfeizhisou middle and low dose groups are significantly decreased, and it is decreased obviously in Xuanfeizhisou middle dose group (P<0.001); Xuanfeizhisou high and Robitussin group compare with model group, no significant difference.Conclusion:Xuanfeizhisou has the function of antitussive and significant reducing the alters of airway inflammation. This side antitussive mechanism may be related to the relief of tracheal inflammation. The method of small dose of LPS induced by simulated gram-negative infections can be successfully created PIC animal model, and it is strong operability; Xuanfeizhisou can reduce neurotransmitter (mainly tachykinin SP, NKA, NKB and CGRP) content, relieve airway inflammation, promote disease recovery, particularly in the middle dose. This study confirmes that traditional Chinese medicine compound Xuanfeizhisou for a good therapeutic effect of cough after infection, and its mechanism is the preliminary study, to provide the experimental basis for the clinical application of the scientific party, and suggested that traditional Chinese medicine compound has potential clinical application.