Integrated metabolomic analysis for intervention effects of Gushudan on glucocorticoid-induced osteoporostic rat plasma based on RP/HILIC-UHPLC-Q-Orbitrap HRMS.

Abstract

An integrated metabolomic strategy based on both RP-UHPLC-Q-Orbitrap HRMS and HILIC-UHPLC-Q-Orbitrap HRMS in rat plasma was developed and validated, to further understand the anti-osteoporosis effect of Gushudan (GSD) and its mechanism on glucocorticoid-induced osteoporosis (GIOP) rats in this study. The metabolites were separated and identified on C18 column (100\u202fmm\u202f×\u202f2.1\u202fmm, 1.7\u202fμm) and Amide column (100\u202fmm\u202f×\u202f2.1\u202fmm, 1.7\u202fμm) using the UHPLC-Q-Orbitrap system (Thermo Fisher Scientific, USA). As a result, a total of 40 differential metabolites were identified, which were mainly related to lipid metabolism, amino acid metabolism, energy metabolism and intestinal flora metabolism. It s worth mentioning that some new potential biomarkers associated with osteoporosis such as 3-hdroxybutyric acid and glycocholic acid were discovered in this study for the first time. With pattern recognition analysis of metabolite profile, a clear separation of the model group and the control group was acquired for plasma samples. The GSD group showed a predisposition towards recovery mimicking the control group, which was in agreement with the behavioral and biochemical results. The present study suggested that GSD had significant anti-osteoporotic effects on glucocorticoid-induced osteoporosis rat plasma, which might be attributed to regulating multiple metabolic pathways. Thus, metabolomics would be a useful tool in the evaluation of the efficacy and elucidation of the mechanism underlying the complex traditional Chinese medicine prescriptions.