UPLC-orbitrap-MS-based metabolic profiling of HaCaT cells exposed to withanolides extracted from Datura metel.L: Insights from an untargeted metabolomics

Abstract

In recent decades, more and more attention to the withanolides extracted from Datura metel.L has been paid due to their anti-psoriatic effects. Withanolides have also been reported to exhibit anti-inflammatory and anti-proliferative properties. Thus, withanolides have been considered as a promising candidate of anti-psoriatic drug. The aim of this study was to investigated the metabolic network of HaCaT cells after exposure to withanolides to identify anti-psoriatic mechanism induced by withanolides on skin cells. In this experiment, our results demonstrated that exposure to withanolides at concentrations beyond 50 μg/mL inhibited cell proliferation and induced cell apoptosis in a dose-dependent manner. In addition, withanolides-induced reactive oxygen species (ROS) generation and mitochondrial depolarization in HaCaT cells. In this research, ultra-high performance liquid chromatography coupled with orbitrap mass spectrometry (UPLC-orbitrap-MS) method was applied to profile metabolite changes in HaCaT cells exposed to withanolides. In total, significant variations in 38 differential metabolites were identified between withanolides exposure and untreated groups. The exposure of HaCaT cells to withanolides at the dose of 200 μg/mL for 24 h was revealed by the disturbance of energy metabolism, amino acid metabolism, lipid metabolism and nucleic acid metabolism. UPLC-orbitrap-MS-based cell metabolomics provided a comprehensive method for the identification of withanolides’ anti-psoriasis mechanisms in vitro. And above metabolic disorders also reflected potential therapeutic targets for treating psoriasis.