Study On The Pathological Mechanism Of Qi Deficiency And Blood Stasis Syndrome In Chronic Heart Failure And Chinese Medicine Intervention Based On Intestinal Microecology And Metabolomic

Objective: To investigate the metabolites and changes in the small intestinal mucosal flora of rats with Qi deficiency and blood stasis evidence of chronic heart failure,as well as the mechanism of action of Yangxin Tongmai formula;to analyze the intestinal flora and metabolite correlation in rats with Qi deficiency and blood stasis syndrome of CHF.Methods:108 SD rats were randomly divided into normal group with 12 rats,sham operation group with 12 rats and model group with 84 rats.The model was established by ligating left anterior descending coronary artery combined with starvation swimming.After 6 weeks,84 rats were randomly divided into 7 groups(n=12): the model group(referred to as the chf group),the Chinese medicine group(referred to as the yxtm group,12g/kg-d)and the western medicine group(referred to as the xst group,0.072mg/kg-d).All 3 groups were gavaged for 4 weeks;model group + normal group feces(referred to as F＿zc group),model group + model group feces(referred to as F＿chf group),model group + Yangxin Tongmai formula group feces(referred to as F＿yxtm group),pseudo-sterile model group + Yangxin Tongmai formula group feces(referred to as Fw＿yxtm group),all 4 groups were gavaged for 2 weeks.Serum NT-pro BNP was measured by ELISA,ileal mucosal flora 16 S r RNA sequencing and plasma metabolite alterations were analysed by LC-MS metabolomics.Results:(1)The content of serum NT-pro BNP in the model group and its fecal transplantation group increased significantly(P < 0.05),while NT-pro BNP in valsartan group,Yangxin Tongmai recipe group and its fecal transplantation group showed a downward trend.(2)16S r DNA sequencing results: Alpha diversity index showed that chao1 index of chf group was significantly lower than sham group(P < 0.05),while chao1 index of yxtm group,F＿zc group and Fw＿yxtm group was significantly higher than chf group(P < 0.05).shannon index was used to measure the homogeneity of the community and there was no significant difference between the groups.There was a decreasing trend in the chf,F＿chf and Fw＿yxtm groups.Beta diversity analysis showed that the chf group,as a faecal donor,had a similar distribution of flora to that of the F＿chf group(P > 0.05),indicating that the faecal transplantation method of gavage with faecal filtrate was successful.compared to the chf and F＿chf groups,the yxtm,F＿xytm,Fw＿yxtm group and F＿zc group were significantly different(P < 0.05)in terms of flora structure.At the phylum level,the phylum Firmicutes,Proteobacteria and Acidobacteria were the top three dominant phyla in terms of abundance of microbiota in the small intestinal mucosa(see Figure 4).There was a significant difference(P < 0.05)in the abundance of the warty microflora between the groups,with the highest abundance levels in the chf group and the lowest levels in the F＿zc group.Analysis to screen for differential groups showed that the sham group was significantly enriched by the dominant bacteria Clostridia,Allobaculum,Erysipelotrichi,Desulfovibrio,the F＿zc group was characterised by the enrichment of the dominant bacteria Sutterella,Alcaligenaceae,The chf group is characterized by the enrichment of dominant bacteria Candidatus＿Arthromitus,and the F＿chf group is characterized by the enrichment of Lactobacillus and Bacilli.the yxtm group was characterized by the enrichment of the dominant bacterium Clostridiaceae,Dorea,Pasteurellales,Aggregatibacter The F＿yxtm group was characterised by a significant increase in the dominant bacteria Prevotellaceae,Akkermansia and Verrucomicrobia.(3)Metabolomic results: Pathway enrichment was performed,showing that metabolic pathways were mainly enriched in glycine,serine and threonine metabolism,caffeine metabolism,galactose metabolism,phenylalanine metabolism,steroid hormone biosynthesis,glycerophospholipid metabolism,arginine biosynthesis,unsaturated fatty acid biosynthesis,porphyrin metabolism,tyrosine metabolism,nitrogen metabolism,carbon metabolism,and cofactor biosynthesis.(4)Intestinal flora and metabolomic correlation: According to Spearman correlation analysis,the results showed that Clostridium,Dorea,Candidatus＿Arthromitus,Desulfovibrionales,Allobaculum,Erysipelotrichi and other differential flora were significantly correlated with differential metabolites.Conclusion:(1)The richness of intestinal flora in rats with CHF syndrome of qi deficiency and blood stasis decreased,and Candidatus＿Arthromitus,Lactobacillus and Bacilli may belong to CHF harmful bacteria.Clostridiaceae,Dorea,Pasteurellales,Aggregatibacter,Prevotellaceae,Akkermansia,Verrucomicrobia and other bacteria may have protective effects on CHF rats with qi deficiency and blood stasis syndrome.(2)Yangxin Tongmai formula can improve CHF through glycine,serine and threonine metabolism,caffeine metabolism,galactose metabolism and other metabolic pathways.(3)This study suggests that there is a complex regulatory network among CHF syndrome of qi deficiency and blood stasis,intestinal flora and plasma metabolic changes.Through Spearman correlation analysis,it is found that Clostridium,Dorea and other bacteria may play an important role in improving CHF syndrome of qi deficiency and blood stasis through influencing metabolic pathways.