An Experimental Study On The Mechanism Of The Function Of The Fei-Xi-Tiao-Zhi-Fang Based On The Intestinal Microenvironment

Objects:In this experiment,we observed the regulation of lung and intestine microflora and mucosal immune molecules sIgA in the model rats by observing the clinical prescription of the famous traditional Chinese medicine in Yunnan to treat children with recurrent respiratory infection,and analyzed the correlation of their correlation.From the angle of the intestinal microflora,the biological basis of "regulating lung system" in traditional Chinese medicine was preliminarily clarified.Methods:1.Group: SPF Wistar rats,male,body weight 100 ± 10 g,according to experimental needs were randomly divided into normal control group,model control group,pulmonary modulation（high,medium,low）concentration group and positive drug group.2.Establishment of bacterial population and immune dysregulation model: A bacterial model of immune flora and immune disorders was established by intraperitoneal injection of a mixture of cephradine,gentamycin sulfate and dexamethasone injections for eight consecutive days.3.Chinese medicine oral drug intervention: The rats in the low,medium and high concentration groups were given different doses of decoction orally every 24 h.Meanwhile,the positive control group were given Broncho-Vaxom with equivalent dose and the normal control and model control groups were given the same amount of11 normal saline in the same way.The above operation was once a day for eight days in a row.4.Index observations: Changes in behavior of rats were observed.Rats were sacrificed,spleen and thymus were calculated for organ index;sIgA in lung and gut were detected by enzyme-linked immunosorbent assay;the microbiota was sequenced by 16 S r DNA;the content of short-chain fatty acids in the intestine of rats was detected by gas chromatography.Results:1.Changes in behaviorIn the model control group,the diet was reduced,the hair lost luster,tired and curled,cold,cyanosis of lips,dilute stool,unformed feces,and symptoms of sneezing and the increase of mouth and mouth secretions.The above symptoms were relieved in the positive control group compared with the model control group.The intervention group of lung system treatment group was relieved than the model control group.2.Changes in thymus and spleen indexThe thymus index and spleen index of the model group decreased significantly compared with blank group,the difference was statistically significant（P<0.05）.The thymus and spleen index of the high concentration group increased significantly,compared with the model group,the difference was statistically significant（P<0.05）.Compared with the model group,the thymus index in the concentration group increased significantly,and the difference was statistically significant（P<0.05）,and the spleen index in the concentration group showed an upward trend.The thymus index of low concentration group was significantly increased,compared with the model group,the difference was statistically significant（P<0.05）,and the spleen index of low concentration group was on the rise.The thymus and spleen index of the positive drug group increased significantly,compared with the model group,the difference was statistically significant（P<0.05）.3.Changes in lung and intestine sIgAThe expression level of sIgA in lung tissues of model group was significantly12 lower than that in the normal control group,and the difference was statistically significant（P<0.01）.Compared with the model group,the pulmonary sIgA of the high concentration group was significantly higher than that in the model group（P<0.01）.The lung sIgA in the concentration group was significantly increased,and the difference was statistically significant compared with the model group（P<0.05）.Compared with the model group,the lung sIgA in the low concentration group increased,and the lung sIgA in the positive drug group increased obviously.Compared with the model group,the difference was statistically significant（P<0.05）.Compared with the the normal control group,the intestinal sIgA of the model control group decreased significantly（P<0.01）.The intestinal sIgA in the high concentration group was obviously increased,the difference was statistically significant compared with the model group（P<0.05）,and the concentration group increased,compared with the model group,the difference was statistically significant（P<0.05）,and the low concentration group rose,compared with the model group,the difference was statistically significant（P<0.05）;the intestinal sIgA of the positive drug group increased significantly,compared with the model group,the difference was significant（P<0.01）.4.Changes of intestinal short-chain fatty acidsCompared with the normal group,the intestinal short chain fatty acids（acetic acid,propionic acid,isobutyric acid,butyric acid,isovaleric acid and valerate）in the model group decreased.Compared with the model group,the intestinal short chain fatty acids in the Chinese medicine concentration group and the positive drug group were on the rise.5.Changes in lung and intestinal flora in the phylum and genus levelCompared with the normal control group,the intestinal flora Firmicutes and Proteobacteria decreased,Bacteroidetes increased,Ruminococcaceae_UCG-013 increased,and Desulfovibrio,Ruminococcus₁ and Turicibacter decreased in the model group.Compared with the model group,the level of Firmicutes in the rats was increased,the change of Proteobacteria was not obvious,the Bacteroidetes decreased,and the Ruminococcaceae_UCG-013 decreased,and Desulfovibrio,Ruminococcus₁ and Turicibacter were increased.In the positive drug group,Firmicutes decreased,Bacteroidetes and Proteobacteria increased,Ruminococcaceae_UCG-013 decreased,and Desulfovibrio,Ruminococcus₁ and Turicibacter increased.Compared with the normal control group,the lung flora Proteobacteria and Bacteroidetes decreased,Firmicutes increased,Pseudomonas and Prevotella₂decreased,and Ralstonia and Blautia（Blaut S bacteria）increased in the model group.Compared with the model group,the lung flora Proteobacteria and Bacteroidetes increased,Firmicutes decreased,Pseudomonas and Prevotella₂ increased,and Ralstonia and Blautia decreased in the rat model group.In positive drug group,Proteobacteria and Bacteroidetes increased,Firmicutes decreased,Pseudomonas and Prevotella₂ increased,Ralstonia and Blautia decreased.6.Correlation analysisShort chain fatty acids were positively correlated with sIgA in lung and gut mucosa.P_<sub>Firmicutes,g_<sub>Lactobacillus were negatively correlated with sIgA and short chain fatty acids,lung and intestinal mucosa（P<0.05）.Conclusions:1.Fei-Xi-Tiao-Zhi-Fang has a regulatory effect on the diversity of lung and intestinal flora and dominant bacteria;2.Fei-Xi-Tiao-Zhi-Fang promotes the secretion of sIgA in the lung intestinal mucosal immune molecule;3.Fei-Xi-Tiao-Zhi-Fang can improve the balance of intestinal microflora by regulating intestinal microflora and mucosal immunity in rats,thus promoting the balance of respiratory flora and mucosal immunity,which is the biological basis of "regulating lung system".