| Objective:1.The characteristics and rules of bronchial asthma in TCM syndromes should be deeply excavated to provide more effective references for clinical work.2.By observing the m RNA expressions of Bax,c-myc,p-p38 MAPK,p-p38MAPK/p38 MAPK and p38 MAPK in the p38 MAPK signaling pathway of rats with bronchial asthma model under the action of Asthma Granule,the molecular mechanism of the action of Asthma Granule was further explored.Methods:1 Clinical studies:Characteristics of bronchial asthma syndromes learn to sum up,to develop a bronchus asthma clinical syndrome of TCM study questionnaire,into the choice standard of 98 cases of patients with syndromes study data,statistical analysis,inductive factors such as gender,age,cause,living environment,sums up the bronchus asthma in TCM syndrome characteristics and laws.2 Animal experimental studies:Seventy male SD rats were randomly divided into 7 groups: blank group,model group,Asthuping high-dose,medium-dose and low-dose groups,Suhuang Zhike Capsules group,and Guilong Kechuanning group.Ovalbumin(OVA)was used to sensitize and stimulate asthmatic rats.After successful modeling,relevant drugs were given by intragastric gavage.After 3 weeks of intragastric gavage,samples were collected and indicators were detected.Lung function of rats in each group was detected and recorded.The pathological changes of lung tissue in each group were observed by HE staining.The expression levels of Bax and c-myc in airway smooth muscle of SD rats in each group were detected by ELISA.WB method was used to detect the expression levels of p-p38 MAPK and p38 MAPK proteins in the tissue specimens of airway smooth muscle in each group.The level of p-p38 MAPKm RNA in lung tissue of rats in each group was detected by q RT-PCR.Results:1 Clinical studies:The syndrome investigation showed that among the 98 patients with bronchial asthma,24 cases(about 24.49%)had the syndrome of external cold and internal drinking,16cases(about 16.33%)had the syndrome of phlegm turbidity obstructing lung,12 cases(about 12.24%)had the syndrome of wind and phlegm obstruction obstructing lung,14cases(about 14.29%)had the syndrome of phlegm heat obstructing lung,and 6 cases(about 6.12%)had the syndrome of lung qi deficiency.There were 12 cases(about12.24%)of lung-qi deficiency syndrome,5 cases(about 5.10%)of lung-kidney qi deficiency syndrome,3 cases(about 3.06%)of lung-kidney Yang deficiency syndrome,1 case(about 1.02%)of Yang burst deactivation syndrome,and 5 cases(about 5.10%)of blood stasis syndrome.The cases investigated in this study had the largest proportion of the syndrome of cold drinking inside the home and abroad,and the other main syndromes included phlegm and turbidities blocking the lung,wind-phlegm blocking the lung,phlegm and heat obstructing the lung,and the other syndromes accounted for a small proportion.In the 98 cases of bronchial asthma patients,more than 50% of the patients presented symptoms of wheezing,shortness of breath,itching of throat,cough and chills.This is consistent with the distribution of TCM syndromes.2 Experimental study:2.1 Lung function:The lung function of model group was significantly worse than blank group.Compared with model group,lung function indexes of rats in each drug intervention group were improved.In addition,the improvement degree of lung function in Asthuping high-dose group,Suhuang group and Guilong Kechuanning group was significantly better than Asthuping medium and low dose group,the improvement of lung function in Asthuping high-dose group was better than Suhuang group and Guilong Kechuanning group,and there was no significant difference in lung function indexes between Suhuang group andGuilong Kechuanning group.2.2 HE staining results of each group:In the blank group,the lung tissue morphology was basically normal,with relatively clear structure of each layer,no glandular hyperplasia in the submucosa,no obvious infiltration of inflammatory cells in the lung bronchus and its surrounding tissues,no obvious abnormalities in the tracheal mucosal epithelium,no obvious secretions in the lumen,and no obvious stenosis in the airway.Compared with the model group,the lung tissue structure was relatively disorganized,with proliferation and shedding of epithelial cells,and a large number of inflammatory cells could be observed.Compared with the model group,the infiltration of inflammatory cells in the low dose group of Asthmaping decreased.Compared with the low dose group,the lung tissue structure and inflammatory cell infiltration were improved in the medium dose group.Asthuping high-dose group,Guilong Kechuanning group and Suhuang group: Compared with medium and low dose Asthuping group,inflammatory cell infiltration was rare,vascular wall and airway wall were not significantly thickened,and airway epithelial cells were relatively intact.2.3 Detection results of Bax and c-myc in lung tissue:The results showed that the expression level of Bax was higher in the lung tissue of rats in blank group.The expression level of Bax in lung tissue of model group was significantly lower than that of blank group.The expression level of Bax in lung tissue of each drug intervention group was significantly higher than that of model group.The expression level of Bax in lung tissue of rats in Asthuping high-dose group was significantly higher than that in other drug intervention groups.The expression of Bax in lung tissue of rats in Suhuang group was higher than that in medium and low dose asthmaping groups.The expression level of Bax in lung tissue of Guilong Kechuanning group was significantly higher than that in medium and low dose Asthuping groups,and higher than that in Suhuang group.There was no significant difference in Bax expression between Asthma medium dose and low dose groups.The results indicated that the expression level of Bax protein in lung tissue was significantly increased in Asthuping high,middle and low dose groups,Guilong Kechuanning group and Suhuang group,and the improvement was most obvious in Asthuping high dose group.The expression of c-myc in lung tissue of model group was significantly higher than that of blank group.Compared with model group,the content of c-myc in lung tissue of rats in Asthuping high-dose group,Asthuping medium-dose group,Asthuping low-dose group,Suhuang group and Guilong Kechuanning group was significantly decreased.Moreover,the expression of c-myc decreased significantly in Asthuping high-dose group than in Asthuping medium-dose group,Asthuping low-dose group,Guilong Kechuanning group and Suhuang group.The decrease degree of c-myc expression in Suhuang group and Guilong Kechuanning group was better than that in Asthuping medium dose group and Asthuping low dose group,and the decrease degree of c-myc in Guilong Kechuanning group was higher than that in Suhuang group.The results showed that the expression of c-myc in lung tissue was significantly inhibited in Asthuping groups,Guilong Kechuanning group and Suhuang group,and the improvement degree was most obvious in the high dose group.2.4 Detection results of p38 MAPKm RNA in lung tissue:The results showed that compared with the blank control group,the expression of p38 MAPKm RNA was increased in the model group.Compared with the model group,the expression of p38 MAPKm RNA was decreased in each drug intervention group.The effect of reducing p38 MAPKm RNA expression in order from high to bottom was Asthuping high dose group > Guilong Kechuanning group > Suhuang group >Asthuping medium dose group > Asthuping low dose group.2.5 Detection results of lung tissue p-p38 MAPK and p-p38MAPK/p38MAPK:The results showed that compared with the blank control group,the expression of p-p38 MAPK protein was increased in the model group.Compared with the model group,the expression of p-p38 MAPK protein was significantly decreased in each drug intervention group.The effect of reducing p-p38 MAPK in Asthuping high-dose groupand Guilong Kechuanning group was more significant than that in Asthuping medium-dose group,low-dose group and Suhuang group.There was no significant difference in the decrease of p-p38 MAPK protein level between Asthuping high-dose group and Guilong Kechuanning group.There was no significant difference in the decrease of p-p38 MAPK protein level between Asthmping medium dose group and Suhuang group.The ratio of p-p38 MAPK protein to p38 MAPK protein in lung tissue of rats with bronchial asthma in each group was detected by Western Blot.The results showed that compared with the blank control group,p-p38MAPK/p38 MAPK increased in the model group.Compared with the model group,p-p38MAPK/p38 MAPK was significantly reduced in each intervention group.The effect of reducing p-p38MAPK/p38 MAPK in Asthuping high-dose group and Guilong Kechuanning group was more obvious than that in Asthuping medium-dose group,low-dose group and Suhuang group.There was no significant difference in the reduction of p-p38MAPK/p38 MAPK between the Asthuping high-dose group and the Guilong Kechuanning group.There was no significant difference in the reduction of p-p38MAPK/p38 MAPK between the Asthmaping medium dose group and the Suhuang group.Conclusion:1.Through the syndrome study,it was found that in the 98 cases of bronchial asthma patients investigated,the syndrome types were mainly cold internal drinking syndrome,phlegm turbidity blocking the lung,wind-phlegm blocking the lung,phlegm heat obstructing the lung,and lung qi deficiency syndrome.2.According to the results of animal experiments,Asthma Ping Granule can effectively improve lung function and pathological changes of lung,reduce airway remodeling and improve asthma symptoms in rats with bronchial asthma.Its action may include inhibiting the expression level of p-p38 MAPK on the p38 MAPK signaling pathway,thereby reducing airway inflammation.Inhibit the abnormal proliferation of cells and maintain the normal balance of apoptosis.Its targets include inducing the down-regulation of c-myc factor in lung tissue and increasing the level of Bax factor in lung tissue.More comprehensive and specific utility targets still need to be further explored. |
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