| ObjectiveTo observe the effects of Yiqi Huatan Quyu Formula on the immune function and pulmonary-intestinal microbiota in rats of COPD,explore the potential relationship between Yiqi Huatan Quyu Formula in regulating COPD immune function and pulmonary-intestinal microbiota,and provide new ideas for further exploring the mechanism of Yiqi Huatan Quyu Formula in treating COPD and clinical treatment of COPD.Methods24 male SD rats(SPF-grade)were fed adaptively in the animal room for 1 week,and then were divided into control group,COPD model group,and Yiqi Huatan Quyu Formula group.Except for the control group,the other two groups were established COPD models:rats were placed in a self-made water tank and forced to swim for 30minutes every day.Then,rats were placed in a 1m~3fumigation box and smoked with 20cigarettes once a day for 1 hour.Next,rats were placed in an oxygen deficient device for 7 hours.The device automatically regulated the oxygen concentration(10±0.5)%.After 6 days of modeling each week,they were suspended on the 7th day for a total of 4weeks.After the completion of modeling,rats in the control and COPD groups were gavaged with 1ml/100g of physiological saline once a day,while those in the Yiqi Huatan Quyu Formula group were gavaged with 1ml/100g(50mg of Yiqi Huatan Quyu Formula per milliliter)of Qibai Pingfei liquid once a day for 2 consecutive weeks.Observe the general condition of rats every day,and after the experiment,detect the lung function,Th17 cells,Treg cells and their related cytokines,transcription factors,and pulmonary and intestinal microbiota of rats.Results1.Lung function:The FEV 0.3,FVC and FEV 0.3/FVC parameters of rats in the COPD model group were significantly reduced,and the prescription for Yiqi Huatan Quyu Formula can improve lung function.2.Th17 and Treg cells:In the model group,Th17 cells and Th17/Treg levels significantly increased,while Treg cell levels decreased.The Yiqi Huatan Quyu Formula promoted Th17 and Treg cells to reach a new balance.3.Serum cytokines:CCL20,CCR6,and IL-17A increased and IL-10 expression decreased in the model group rats.Yiqi Huatan Quyu Formula can inhibit inflammatory factors and regulate immune factors.4.Transcription factor RORγt and Foxp3:The expression of RORγt increased and the expression of Foxp3 decreased in the lung tissue of model group rats.The Yiqi Huatan Quyu Formula can regulate the transcription factor expression of Th17 and Treg cells.5.Diversity of pulmonary and intestinal microbiota:The species richness of pulmonary and intestinal microbiota was similar among the three groups of rats.The uniformity of pulmonary and intestinal microbiota in the COPD group was lower than that in the control group,and increased after intervention with Yiqi Huatan Quyu Formula.The PCo A diagram showed significant differences in pulmonary and intestinal microbiota among the three groups of rats.6.Microbial community structure of lung and intestine:According to the taxonomy composition analysis,the dominant bacteria under phyla level in the lung and intestine of rats in each group were composed of Bacteroidetes,Proteobacteria and Firmicutes.Among them,Bacteroidetes and Proteobacteria in the lung and intestine of the model group were lower than those of the control group.As for Firmicutes,compared with the control group,it decreased in the lung and increased in the intestine of the model group.7.Difference analysis of pulmonary and intestinal microbiota:The difference analysis of pulmonary microbiota showed that 22 bacterial genera decreased and 3bacterial genera increased in the COPD group than that in the control group;compared with COPD group,13 bacterial genera increased and 8 bacterial genera decreased in the QBPF group.The difference analysis of microbiota in the gut showed that 8 bacterial genera decreased and 10 bacterial genera increased in the COPD group than that in the control group;compared with COPD group,2 bacterial genera increased and 9 bacterial genera decreased in the QBPF group.8.Functional prediction analysis of pulmonary and intestinal microbiota:The functional prediction analysis based on KEGG showed that the lung and intestinal Amino acid related enzymes,Aminoacyl-t RNA biosynthesis,and Purine metabolism pathways were enriched in COPD group,while they were significantly reduced after the intervention of the Yiqi Huatan Quyu Formula.9.Correlation analysis of pulmonary and intestinal microbiota:There were 9 kinds of the same bacterial genera in the lung and intestine of rats,with correlations between them.In addition,other pulmonary and intestinal bacterial genera also had significant associations.10.The correlation between microbiota in the lung and gut and lung function and immune features:Mycoplasma enriched in the lung of COPD group was positively correlated with Th17-related cytokines,while it was negatively correlated with pulmonary function and Treg-related cytokines.Acetatifactor had a lower intestinal abundance in the COPD group,and was positively correlated with lung function and Treg-related cytokines,and negatively correlated with Th17/Treg ratio and Th17-related cytokines.Conclusions1.The Yiqi Huatan Quyu Formula can regulate the lung function parameters of COPD rats and promote a new balance of Th17/Treg.2.Yiqi Huatan Quyu Formula can regulate the composition of pulmonary and intestinal microbiota in COPD rats,improve the community structure,and inhibit the relative abundance of the intestinal coprococcus＿2,Prevotella＿9,and Blautia and Mycoplasma in the lung,and increase the relative abundance of the intestinal Prevotella＿UCG＿003 and the pulmonary Rikenellaceae＿RC9＿gut.3.The correlation analysis between pulmonary and intestinal microbiota further confirmed the lung-gut axis.4.There was a significant correlation between lung function and immune status and pulmonary and intestinal microbiota in COPD rats. |
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