| Objective: To find the potential biomarkers and metabolic pathways related to the treatment of calcium oxalate kidney stones by Desmodii Styracifolii Herba(DSH),and to identity and compare the metabolites of schaftoside,an active ingredient in DSH,in healthy rats and calcium oxalate kidney stones rats in vivo and in vitro.Methods: 40 male SD rats were randomly divided into 5 groups(n = 8): the blank group,the model group,the low-dose treat group,the medium-dose treat group and the high-dose treat group.After modeling and treatment,the urine,serum and kidney tissues of each group rats were collected,and the urine smears and renal tissue sections were observed under a microscope.Firstly,the UHPLC-Q-TOF-MS/MS technology was used to collect the information date of endogenous compounds in serum samples,and Progenesis QI and SIMCA-P 14.1 software were used to process and analyze the date.Compounds that satisfied both P<0.05 and VIP>1 were considered as differential compounds between different groups and the differential metabolites were identified.Then,the heat map and related metabolic pathways were processed by an online platform Metabo Analyst 4.0 and the KEGG database,and the potential biomarkers associated with the therapeutic effects of DSH were screened.Finally,the Cytoscape software was used to construct a metabolite-gene-protein biological information network diagram.The UHPLC-Q-TOF-MS/MS technique and the metabolite analysis strategy were used to identify the structures of schaftoside metabolites in plasma,urine,feces,bile and intestinal bacteria samples of the two kind rats.And the metabolic pathways of schaftoside in two kinds of rats were also compared.In addition,the relative quantitative method was used to find the difference of metabolite content in the two kinds of rats.Results: A total of 72 differential metabolites were identified between the kidney stone model group rats and the healthy group rats.88 differential metabolites were identified between the DSH treatment groups and the kidney stone model group.The metabolic pathways involved mainly include sphingolipid metabolism,phenylalanine metabolism,pantothenate and Co A biosynthesis,and arachidonic acid metabolism.A total of 5 compounds: phytosphingosine,sphingasine,Cer(d18:0/16:0),L-phenylalanine and pantothenic acid were screened out as potential biomarkers associated with the therapeutic effects of DSH.By constructing bioinformatics network,78 key genes and 50 related proteins were found.A total of 28 metabolites were found in healthy rats and 30 metabolites in model rats after oral administration of schaftoside.The main metabolic pathways included de-glycosylation,oxidation reaction,reduction reaction,acetylation reaction,internal hydrolysis,methylation reaction,glucuronide conjugation and sulfate conjugation.The relative peak area method was used to compare the differences of the metabolites in these two kinds of rats.The results showed that there were differences in metabolite types and contents between healthy rats and model rats.Conclusion: In this study,based on the UHPLC-Q-TOF-MS/MS and metabolomics method,the potential biomarkers and metabolic pathways related to the therapeutic effects of DSH were found,and a comprehensive biological network from metabolites to genes and proteins was constructed.These research results provide a comprehensive understanding for the study of the biological mechanism of DSH in the treatment of kidney stones.In addition,a metabolite analysis strategy was used to identify the metabolites of schaftoside in healthy rats and calcium oxalate kidney stones rats,and the metabolites of schaftoside in the two kinds of rats were compared from two levels: metabolites type and metabolites content.The study of the metabolism in model rats can help to understand the metabolic process of schaftoside in clinical practice. |
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