The Research Of The Therapeutic Effect Of WentongFang On Allergic Asthma Rat From The Perspective Of Intestinal Flora

Objective:To study the therapy effect of WentongFang on allergic asthma rat model from the changes of general state,airway responsiveness,total serum IgE of peripheral blood,pulmonary pathomorphology,gut microbiota structure,in order to confirm the existence of intestinal flora imbalance in allergic asthma rat model and the the WentongFang probably can cure allergic asthma by the affect of intestinal flora.Methods:40 rats were randomly divided into four groups(normal group,model group,dexamethasone group,WentongFang group),and we fed them for a week.Sensitized them with injection of a mixture of OVA/Al(OH)3 and 1ml 0.9%saline in five point(four palms and abdomen),and 0.2ml DPT vaccine in abdomen at the same time(in the first days and eighth days,a total of 2 times),and then exposed them in 1%OVA aerosol inhalation(from fifteenth days,once a day,20 min/times,a total of 7 times),and we got the allergic asthma model.Then the rats were given gavage administration for 14 days.WentongFang group were given the extractum of WentongFang in dosage of 1.34g/kg,dexamethasone group were given 0.81mg/kg dexamethasone solution.The other groups were given the same amount of 0.9%saline.Then we observed generally condition of the rats,measured their airway responsiveness,collected the peripheral blood for total serum IgE,collected the lung tissue for HE staining analysis for assessing lung injury in rats,and collected the cecal feces for 16S rDNA sequencing analysis to observe the changes of intestinal flora.Results:1.General condition:After modeling,all the rats were showed typical symptoms of allergic asthma.After gavage,the symptoms of asthma model group was the most significant,compared with the model group,dexamethasone group,Wentongfang group symptoms significantly reduced.2.HE staining of lung tissue:no inflammatory cell infiltration was found in the normal group,no bronchospasm,and the structure of the alveolar and bronchial mucosal epithelium was clear and complete.Both the model group,dexamethasone group and Wentongfang group have typical allergic asthma pathological manifestations,just like the inflammatory cell infiltration in lung tissue,bronchial epithelial cells arranged in disorder and flake off,alveolar septal thickening and bronchospasm,However,the degree of pathological damage is different,the order of the degree of pathological changes from high to low is model group,wentongfang group,dexamethasone group,normal group.3.Determination of airway responsiveness:There were significant differences in the basal expiratory resistance and the expiratory resistance at each Ach concentration in model group compared with that in blank group(P<0.05);Compared with the model group,WentongFang group and dexamethasone group have a significant difference in expiratory resistance when the Ach concentration is 50 and 100ug/kg(P<0.05);In each Ach concentration,WentongFang group and dexamethasone group’ bronchial responsiveness showed no significant difference.4.Blood EOS count:Compared with the blank group,there was a statistically significant difference in WentongFang group,dexamethasone group and model group(P<0.07);Compared with the model group,there was a statistically significant difference in WentongFang group and dexamethasone group(P<0.01);Compared with the dexamethasone group,there was a statistically significant difference in WentongFang group(P<0.01).5.Intestinal microflora structure:Alpha diversity:Chao1 index showed that,compared with the model group,there was statistical significance in blank group,dexamethasone group and WentongFang group(P<0.05);Shannon index and Simpson index showed that there was no significant difference between the groups,(P>0.05).Beta diversity:The intestinal microflora structure in dexamethasone group has a maximum difference with other groups and WentongFang group’ is closest to the blank group.Analysis of the difference of bacterial community structure:Compared with the blank group,the Akkermansia,Oscillospira,Coprococcus,Phascolarctobacterium,Roseburia,Prevotella abundance increased in the model group(Ratio kb/mx<0.5),Dorea abundance decreased(Ratio kb/mx>2);Compared with the model group,Phascolarctobacterium,Prevotella and Dorea abundance increased in dexamethasone group(Ratio dex/mx>2),Akkermansia,Oscillospira abundance decreased(Ratio dex/mx<0.5);Compared with the model group,Coprococcus,Phascolarctobacterium abundance increased in WentongFang group(Ratio zy/mx>2),Akkermansia,Oscillospira,Roseburia,Prevotella abundance decreased(Ratio zy/mx<0.5);Compared with the blank group,Akkermansia,Coprococcus,Phascolarctobacterium,Prevotella abundance increased in dexamethasone group(Ratio dex/kb>2),Lactobacillus abundance decreased(Ratio dex/kb<0.5);Compared with the blank group,Coprococcus,Phascolarctobacterium abundance increased in WentongFang group(Ratio zy/kb>2),and there was no significant difference with the blank group in remaining strains(0.5<Ratio zy/kb<2);Compared with the dexamethasone group,Coprococcus abundance increased in WentongFang group(Ratio dex/zy>2),Akkermansia,Phascolarctobacterium,Roseburia,Prevotella,Dorea abundance decreased(Ratio dex/zy<0.5).Conclusion:1.Preliminary confirmed the WentongFang has a beneficial effects in general condition,airway reactivity,Blood EOS count and lung tissue pathological.That the party has the effect of relieving symptoms,reduce airway responsiveness,reduce airway inflammation of rats with allergic asthma.2.Preliminary confirmed the the rat model of allergic asthma exist the imbalance of intestinal flora.After the treatment of the intestinal flora construction of dexamethasone group was different from blank group,which indicated that the treatment effect of dexamethasone in allergic asthma could not be explained by adjusting the structure of intestinal bacterial.After treatment,the symptom and related index of Wentongfang group was relieved,at the same time,the structure of intestinal bacterial was also changed,it’s more closed to blank group compared with the dexamethasone group,which showed that,to some extent WentongFang may relieves allergic asthma symptoms and related indexes by the impact to intestinal bacterial structure.