The Effectsof Yi Gong Powder Preventionon The Intestinal Flora-Short Chain Fatty Acid-Treg Cellsin Model Of Micewith Bronchial Asthma

Objective:In this paper,ovalbumin(OVA)-induced young mice are used as animal models,combined with 16S-rRNA sequencing,bioinformatics analysis,gaschromatography/mass spectrometry(GC-MS/MS)detection,and molecular biology techniques to study the effect of early intervention by Yigong powder on the intestinal flora-short-chain fatty acid-regulatory T-cell mechanism inasthmatic model mice,it provides an experimental basis for further exploring the mechanism of early intervention of traditional Chinese medicine"strengthening earth to generate metal" treatment method to prevent and treat children’ s bronchial asthma.Methods:32 female BALB/c mice aged 7 days were randomly divided into normal group,model group,probiotics group(Clostridium butyricum)and Yigong powder groups.After ovalbumin sensitized asthma mice models were successf’ully prepared,the general behavioral activities of these groups will be observed.With 24 hours after the last atomization,the colon stool,serum and lung tissues from all the mice were sacrificed.16S-rRNA sequencing combined with bioinformatics to analyze intestinal flora characteristics in feces;GC/MS technique to detect the concentration of acetic acid,propionic acid and butyric acid in serum;RT-qPCR technique to detect expression for the lung tissue transcripition factor Foxp3、Ffar2 mRNA;Western blot to detect the expression of GPR43 protein in lung tissue.Results:1.The effect of Yigong powder on intestinal flora of colonic fecesThere was a significant difference between the intestinal flora structure of the young mice in the Yigong powder group and the model group,the abundance of the Bacteroidetes in the Yigong powder group was significantly higer than in the model group,while the abundance of Firmicutes is reduced was reduced.2.The effect of Yigong powder on the concentration of acetic acid,propionic acid and butyric acid in serum(1)The serum acetic acid concentration of the model group was higher than that of the blank group,but there was no significant difference.The serum acetic acid concentration values of the Yigong powder group and the probiotic group were higher than the model group,but there was no significant difference;while the relative ratios of serum acetic acid in the Yigong powder group and the probiotic group were significantly higher than those in the model group(P=0.035,0.020,respectively).(2)The serum propionate concentration in the model group was lower than that in the blank group,but there was no significant difference.The serum propionate concentration in the Yigong powder group and the probiotic group was significantly lower than that in the model group(P=0.036,0.027,respectively);the relative ratio of serum propionic acid in the Yigong powder group and the probiotic group was lower than that in the model group,but there was no significant difference.(3)The serum butyric acid concentration in the model.group was lower than that in the blank group,but there was no significant difference.The serum butyric acid concentration in the Yigong powder group and the probiotic group was lower than that in the model group,but there was no significant difference;the relative ratio of serum butyric acid in the Yigong powder group was lower than that in the model group.but there was no significant difference,while the relative ratio of serum butyric acid in the probiotic group was significantly lower than that in the model group(P=0.015).3.The effect of Yigong powder on the expression of transcripition factor Foxp3、Ffar2 mRNA in the lung tissue(1)The relative expression of Foxp3 mRNA in the lung tissue of the model group was higher than that in the blank group,but the difference was not significant.The median value of Foxp3 mRNA expression in the Yigong powder group was the same as that in the model group,and it was not statistically significant.The expression level of the probiotic group was lower than that of the model group.But the difference is not significant.(2)The relative expression of Ffar2mRNA in the lung tissue of the model group was lower than that in the blank group,but there was no significant difference.The relative expression of Foxp3 mRNA in the lung tissue of the Yigong powder group and the probiotic group was higher than that in the model group,but there was no significant difference.4.The effect of Yigong powder on the expression of GPR43 protein in lung tissueThe relative expression of GPR43 protein in the lung tissue of the model group was higher than that in the blank group,but there was no significant difference.The relative expression of GPR43 protein in the lung tissue of the Yigong powder group was significantly higher than that in the model group(P=0.021).the relative expression of GPR43 protein in the lung tissue of the probiotic group was lower than that of the model group,but there was no significant difference.ConlclusionsThe early intervention of Yigong powder in the asthmatic model mice may be related to the regulation of the metabolism of short-chain fatty acids in the intestinal flora,but may not be through the Treg pathway.