Integrated metabolomic and lipidomic analysis revealed the protective mechanisms of Erzhi Wan on senescent NRK cells through BRL cells

Abstract

Ethnopharmacological relevance: Erzhi Wan (EZW), as a prescription of traditional Chinese medicine, has been used for tonifying the liver and kidney. Although past studies have shown that EZW has potential anti-aging effect, the mechanisms associated with cellular metabolomics and lipidomics are not fully understood.

Aim of the study: This study aimed to evaluate the anti-aging effect of EZW and investigate the mechanisms associated with cellular metabolomics and lipidomics.

Materials and methods: EZW solution at dosage of 3.6 g/kg in Sprague-Dawley rats was orally administered twice a day for 7 days and serum containing EZW was then collected. NRK cell senescence model induced by D-galactose was established in vitro, and non-contact co-culture cell assay was performed between senescent NRK cells and BRL cells intervened by serum containing EZW. The anti-aging effect of EZW on NRK cells was evaluated by metabolites identification, differential metabolites screening and metabolic pathways analysis through cellular metabolomics with GC-MS and lipidomics with UHPLC-Q-Exactive Orbitrap/MS.

Results: Serum containing EZW indicated a protective effect through intervening BRL cells in non-contact co-culture system with D-gal-induced senescent NRK cells. For metabolic profiles, 71 endogenous metabolites were identified, among which 24 significantly differential metabolites were screened as metabolomics potential biomarkers. For lipidic profiles, 64 lipid components were identified in NRK cell samples under positive ion mode, among which 24 potential biomarkers of lipids were screened, mainly including PC and PE. 127 lipid components were identified in NRK cell samples under negative ion mode, among which 59 potential biomarkers of lipids were screened, including FA, PC, PE, PI and PS. Metabolic pathway analysis demonstrated that the identified differential metabolites found mainly involved in amino acids metabolism, energy metabolism and phospholipid biosynthesis pathways.

Conclusion: Serum containing EZW exhibited protective effect on D-gal-induced senescent NRK cells through intervening BRL cells by mainly regulating amino acids metabolism, energy metabolism and phospholipid biosynthesis pathways to possess its anti-aging function, providing a theoretical basis for clinical treatment of EZW.

Keywords: Anti-Aging; Erzhi Wan; Lipidomics; Metabolomics; Non-contact co-culture.