Kidney-tonifying mechanism of Gushudan based on UPLC-Q-Orbitrap HRMS metabolomics and 16S rRNA gene sequencing
Objective To investigate the kidney-tonifying mechanism of Gushudan(GSD)in kidney-yang deficiency syndrome(KYDS)rats induced by hydrocortisone by integrating metabonomics and intestinal microbiome.Methods Untargeted metabolomics of UPLC-Q-Orbitrap HRMS and intestinal microbiome analysis of 16S rRNA gene sequencing were used to analyze fecal metabolite profiles and intestinal microflora composition of rats in control group(CON),model group(MOD),GSD treatment group(GSD)and positive group(P).Pearson correlation analysis was used to explore the correlation between endogenous differential metabolites and differential microflora.Results A total of 22 different metabolites,such as tryptophan,chenodeoxycholic acid,creatinine and oleamide,were obtained by UPLC-Q-Orbitrap HRMS in positive and negative ion mode,which were mainly involved in amino acid metabolism,bile acid metabolism,energy metabolism and lipid metabolism.Based on 16S rRNA gene sequencing analysis,GSD significantly regulated the composition of intestinal flora in rats with KYDS,such as significantly up-regulated the relative abundance of Prevotellaceae(P<0.05)and significantly down-regulated the relative abundance of Oscillibacter at the genus level(P<0.05).The results of correlation analysis showed that fecal metabolites of glycocholic acid and chenodeoxycholic acid were significantly positively correlated with Prevotellaceae(P<0.05),but significantly negatively correlated with Phascolarctobacterium(P<0.05),which changed significantly at the genus level.There was a significant negative correlation between docosahexaenoic acid and Lachnospiraceae(P<0.05).Conclusion GSD plays an important role of tonifying kidney by regulating the endogenous metabolism and intestinal microflora structure,providing new ideas and methods for traditional Chinese medicines to treat diseases through the gut-kidney axis.
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