Abstract
Carboxyl-containing metabolites, such as bile acids and fatty acids, have many important functions and microbiota is involved in the production of them. In the previous study, we found that the chronic kidney disease (CKD) model mice raised under germ-free conditions provided more severe renal damage than the mice with commensal microbiota. However, the precise influence by the microbiome and carboxyl-containing metabolites to the renal functions is unknown. In this study, we aimed to develop a novel chemical isotope labeling-LC-MS/MS method using the 2-picolylamine and its isotopologue and applied the analysis of effects of microbiome and CKD pathophysiology. The developed semi-quantitative method provided the high accuracy not inferior to the absolute quantification. By comparing of four groups of mice, we found that both microbiota and renal function can alter the composition and level of these metabolites in both plasma and intestine. In particular, the intestinal level of indole-3-acetic acid, short-chain fatty acids and n-3 type of polyunsaturated fatty acid, which play important roles in the endothelial barrier function, were significantly lower in germ-free conditions mice with renal failure. Accordingly, it is suggested these metabolites might have a renoprotective effect on CKD by suppressing epithelial barrier disruption.
Highlights
Carboxyl-containing metabolites, such as bile acids and fatty acids, have many important functions and microbiota is involved in the production of them
Sajiki et al have reported that post-synthetic H–D exchange methods for various organic compounds in D2O under an atmosphere of H2 in the presence of palladium-on-carbon (Pd/C) and platinum-on-carbon (Pt/C)[35,36], and we applied it for the synthesis of 2PA-d6
We revealed that most bile acid levels in plasma were significantly elevated in specific pathogen-free (SPF)-renal failure (RF) mice compared to control mice, as well as the intestinal level of chenodeoxycholic acid (CDCA), HCA, and DCA were increased (Fig. 5A)
Summary
Carboxyl-containing metabolites, such as bile acids and fatty acids, have many important functions and microbiota is involved in the production of them. The intestinal level of indole-3-acetic acid, short-chain fatty acids and n-3 type of polyunsaturated fatty acid, which play important roles in the endothelial barrier function, were significantly lower in germ-free conditions mice with renal failure It is suggested these metabolites might have a renoprotective effect on CKD by suppressing epithelial barrier disruption. To perform the comprehensive and accurate analysis of carboxyl-containing metabolites for plasma and intestinal samples in CKD model mice with or without the conditions of germ-free, we developed a CIL-LC-MS/MS method for carboxylic acid tagging with 2-picolylamine (2PA) and 2-picolylamine-d6 (2PA-d6). The developed CIL-LC-MS/MS-based approach was successfully used for comprehensive relative quantification of carboxyl-containing metabolites in plasma, feces, and cecal contents from mice and suggested that the novel features related to the microbiome and CKD pathology
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