| Objective: In this study,Yiqi Ditan Poyu Formula is an effective prescription for the treatment of chronic obstructive pulmonary disease by Professor Qingsheng Li,a famous doctor of Traditional Chinese medicine in Yunnan Province,by observing the changes in the contents of inflammatory factors IL-8 and TNF-?,the number of immature dendritic cells in the lung tissue,the lung-intestinal mucosal immune molecule s Ig A,the bacteria flora and the shortchain fatty acids in the lung-intestinal mucosa of the model rats,to preliminarily investigate the biological basis of this Formula for regulating the lung-intestinal microecological balance in rats with COPD,the purpose is to provide theoretical basis for the clinical application of this prescription.Methods: 1.The rat model of COPD was replicated: The method of adding lipopolysaccharide and cigarette smoke by tracheal drip was adopted,except for the blank group,the rats were fed LPS on day 1 and day 14 of modeling,continue to inhale cigarette smoke on days 2-13 and 15-56,twice a day for 30 minutes.2.Drug intervention: After the modeling(Day 57),the low,medium and high dose groups were given the decoction of Yiqi Ditan Poyu Formula by gavage for 28 days,the model group was given the same amount of normal saline.3.Indicator detection:(1)Inflammatory related correlates: The pathological changes of lung tissues and colon tissues were detected by HE staining;changes of IL-8 and TNF-? in rat lung tissues were detected by q PCR;(2)Immunity related indicators: To calculate the change of thymus and spleen index by weighing;the contents of immature dendritic cells in rat lung tissues were determined by immunohistochemistry;s Ig A content in lung and colon tissues was detected by ELISA;(3)Lung-gut microecological related indicators: highthroughput sequencing of 16 S r RNA gene was used detect the abundance and diversity of pulmonary and intestinal flora in rats;the contents of short-chain fatty acids in rat intestine were determined by gas chromatography.Result: 1.Changes in inflammatory markers 1.1 Pathological changes in lung tissue: Compared with the blank group,a large number of inflammatory cells infiltrated around the bronchi and some alveolar stenosis were observed in the model group;compared with the model group,the number of inflammatory cell infiltration in the drug intervention group was reduced,the bronchial structure was relatively intact.1.2 Pathological changes of colon tissue: Compared with the blank group,the rats in the model group could see partial mucosa abscission,edema,muscle layer thinning and inflammatory cell infiltration;compared with the model group,the inflammatory cell infiltration in the drug intervention group was improved,edema was alleviated,and the mucosa was more complete.1.3 Changes of IL-8 and TNF-? in lung tissues: Compared with the blank group,the content of IL-8 in the lung tissue of the model group increased significantly(P <0.05),compared with the model group,the content of IL-8 in the drug intervention group showed a downward trend(P > 0.05).Compared with the blank group,the content of TNF-? in the lung tissue of the model group increased significantly(P <0.01),compared with the model group,the content of TNF-a in the drug intervention group decreased(P <0.05).2.Changes in immune-related indicators 2.1 Changes of thymic spleen index: Compared with the blank group,the thymus index of the rats in the model group decreased significantly(P <0.05),compared with the model group,the thymus index of the rats in the drug intervention group had an upward trend(P > 0.05).Compared with the blank group,the spleen index of the rats in the model group decreased significantly(P <0.05),compared with the model group,the spleen index of the rats in the drug intervention group had an upward trend(P > 0.05).2.2 Changes in the number of immature dendritic cells in lung tissue: Compared with the blank group,the number of immature dendritic cells in the lung tissue of the model group increased significantly(P <0.05),compared with the model group,the number of immature dendritic cells in the lung tissue of the drug intervention group decreased(P > 0.05).2.3 Changes of immune molecule s Ig A in lung tissue: Compared with the blank group,the immune molecule s Ig A in lung tissues in the model group was significantly decreased(P <0.001),while that in the drug intervention group was increased(P <0.05).2.4 Changes of immune molecule s in colon tissue: Compared with the blank group,the immune molecule s Ig A in the colon tissue of the model group decreased significantly(P <0.0001),compared with the model group,the immune molecule s Ig A in the colonic tissues of the rats in the drug intervention group had an upward trend(P > 0.05).3.Changes of related indexes of lung-intestinal microecology 3.1 Changes of pulmonary bacterial flora:(1)Changes in abundance: In terms of the level of bacteriophyta,the rats in the model group Firmicutes(8.38%),Bacteroidetes(13.43%),and Proteobacteria(0.80%)has decreased,Tenericutes(22.24%)has creased,compared with the model group,the content of Firmicutes in the drug intervention group decreased,the content of Tenericutes increased,and there was no significant change in Bacteroidetes and Proteobacteria.In terms of changes in the level of bacteria,the relative abundance of bacteria such as Roseburia(1.92%) decreased significantly after modeling(P <0.05),increased after drug intervention,the relative abundance of bacteria such as Clostridium sensu stricto(4.91%)increased(P > 0.05),and decreased after drug intervention.(2)Diversity change: Compared with the blank group,the lung bacterial diversity of the model group was increased;Compared with the model group,the lung bacterial diversity of the drug intervention group was decreased.3.2 Changes of intestinal flora:(1)Abundance change: In terms of the level of bacteriophyta,compared with the blank group,compared with the blank group,the contents of Firmicutes(3.94%)and Bacteroidetes(6.02%)in the model group decreased,and the contents of Euryarchaeota(5.92%)and Proteobacteria(1.55%)in the model group decreased,compared with the model group,the contents of Firmicutes and Bacteroidetes of the rats in the drug intervention group increased,while the contents of Euryarchaeota and Proteobacteria decreased.In terms of changes in the level of bacteria,the relative abundance of bacteria such as Oscillibacter(5.66%)decreased significantly after modeling(P <0.05),and increased after drug intervention,relative abundance of bacteria such as Escherichia / Shigella(4.42%)increased(P <0.05),and decreased after drug intervention.(2)Diversity change: Compared with the blank group,the diversity of intestinal flora of the model group rats decreased;Compared with the model group,the diversity of intestinal flora of the drug intervention group rats increased.3.3 Correlation analysis of lung flora and intestinal flora: After modeling Alloprevotella,Oscillibacter,Roseburia and Lactobacillus,the relative abundance of pulmonary and intestinal flora decreased,and increased after drug intervention,which were probiotics significantly related to this model.Fusobacterium,Escherichia/Shigella and Clostridium sensu Stricto express the relative abundance of pulmonary bacteria and intestinal bacteria all increased after being modeled,and all decreased after drug intervention.They are all harmful bacteria significantly related to this model.3.4 Changes in short-chain fatty acid content: Compared with the blank group,the content of acetic acid and n-butyric acid in the model group decreased,while that of propionic acid decreased(P <0.01),the propionic acid content showed an decreased trend(P> 0.05);compared with the model group,the drug The content of acetic acid,propionic acid and n-butyric acid in the intestinal contents of the rats in the intervention group all showed an upward trend(P> 0.05).Conclusion: 1.Yiqi Ditan Poyu Formula can reduce the level of pulmonary inflammation in COPD model rats.2.Yiqi Ditan Poyu Formula interrupts disease progression by regulating lungenterobacteria and intestinal metabolites in COPD model rats,as well as improving immunity,restoring its microecological balance. |
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