Investigation of the mechanisms of Genkwa Flos hepatotoxicity by a cell metabolomics strategy combined with serum pharmacology in HL-7702 liver cells

Abstract

1. To investigate Genkwa Flos hepatotoxicity, a cell metabolomics strategy combined with serum pharmacology was performed on human HL-7702 liver cells in this study. 2. Firstly, cell viability and biochemical indicators were determined and the cell morphology was observed to confirm the cell injury and develop a cell hepatotoxicity model. Then, with the help of cell metabolomics based on UPLC-MS, the Genkwa Flos group samples were completely separated from the blank group samples in the score plots and seven upregulated as well as two down-regulated putative biomarkers in the loading plot were identified and confirmed. Besides, two signal molecules and four enzymes involved in biosynthesis pathway of lysophosphatidylcholine and the sphingosine kinase/sphingosine-1-phosphate pathway were determined to investigate the relationship between Genkwa Flos hepatotoxicity and these two classic pathways. Finally, the metabolic pathways related to specific biomarkers and two classic metabolic pathways were analyzed to explain the possible mechanism of Genkwa Flos hepatotoxicity. 3. Based on the results, lipid peroxidation and oxidative stress, phospholipase A/lysophosphatidylcholine pathway, the disturbance of sphingosine-1-phosphate metabolic profile centered on sphingosine kinase/sphingosine-1-phosphate pathway and fatty acid metabolism might be critical participators in the progression of liver injury induced by Genkwa Flos.

The hepatoprotective effect and mechanism of lotus leaf on liver injury induced by Genkwa Flos

“…[8] Both animal experiments in vivo and cell experiments in vitro indicated that excessive or long-term use of the raw Genkwa Flos could result in serious damage to liver. [9,10] As a representative Chinese traditional medicine causing drug-induced liver injury, Genkwa Flos was selected to be used as a moulding agent which induce the liver injury of rats in this study.…”

“…Similarly, the levels of several endogenous metabolites might change when liver suffers from injury. According to the relative literatures, [10,[12][13][14] 5-oxoproline, phenylalanine, tryptophan, C 18 -phytosphingosine, LPC (16 : 0) and LPC (18 : 1) were found to be strongly associated with liver injury. As a significant intermediate of the glutathione synthesis pathway, 5-oxoproline is considered as a potential biomarker for excessive depletion of glutathione and oxidative stress.…”

Discovery of potential therapeutic targets for non-small cell lung cancer using high-throughput metabolomics analysis based on liquid chromatography coupled with tandem mass spectrometry

“…There is a major challenge in the concurrent extraction, separation, and detection of cells because the endogenous metabolites vary widely in their physical and chemical properties in comprehensive characterization of the cell metabolome. 23,24 The chemical properties of a great number of metabolites are still unknown, leading to some difficulty in the problem of standardizing quenching and extraction methods. Some standard compounds are not yet readily available.…”

Investigation of the detoxification effect of licorice on Semen Strychni-induced acute toxicity in rats using a HPLC-Q-TOF/MS-based metabolomics approach

“…Recent years, a number of studies have been conducted to explore the metabolic mechanisms of TCMinduced nephrotoxicity and hepatotoxicity based on metabolomics strategies, and the satisfactory results indicated the applicability of metabolomics strategies. 5,12 Selecting an appropriate analytical platform is critical to the results. Currently, metabolomic analytical platforms used in the majority of studies are nuclear magnetic resonance (NMR), liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS).…”