A combined metabolomics and molecular biology approach to reveal hepatic injury and underlying mechanisms after chronic l-lactate exposure in mice
This research aimed to understand more about whether chronic l-lactate exposure may affect the peripheral tissues of rodents and to look for the underlying pathogenesis. Herein, male C57BL/6 rodents were split into control and l-lactate groups. After l-lactate strategy to eight days (1 g/kg), metabolic alterations in liver, kidney, muscle, and serum samples were based on 1H nuclear magnetic resonance (1H NMR)-based metabolomics. Furthermore, organ function was evaluated by serum biochemical and histopathological examinations. Reactive oxygen species (ROS) levels were measured using dihydroethidium staining amounts of signals involved with lactate metabolic process and ROS-related pathways were detected using western blotting or polymerase squence of events. Apoptosis was detected by TUNEL-fluorescence staining. Metabolomic analysis says l-lactate rodents demonstrated decreased amounts of glutathione (GSH), taurine, ATP, and elevated glucose content, when compared with control rodents. In addition, l-lactate rodents presented considerably greater serum amounts of alanine aminotransferase and aspartate aminotransferase and elevated glycogen content in hepatic tissues, when compared with control rodents. l-lactate rodents also had more apoptotic nuclei within the livers than controls. Furthermore, l-lactate exposure reduced mRNA and protein amounts of superoxide dismutase-2 and c-glutamylcysteine ligase, elevated amounts of cytochrome P450 2E1 and NADPH oxidase-2, and elevated the protein expressions of LDHB, Bax/Bcl-2, cleaved caspase-3, and sirtuin-one in hepatic tissues. Together, these results indicate that chronic l-lactate exposure increases oxidative stress, Dihydroethidium and apoptosis in hepatocytes via upregulation of Bax/Bcl-2 expression and also the consequent mitochondrial cytochrome-C release and caspase-3 activation, which plays a role in the pathogenesis of hepatic disorder.