In vitro toxicological investigation of Fusarium graminearum toxins in Rattus norvegicus myocardial H9c2 cells

Abstract

Mycotoxins are toxic secondary metabolites of fungi that grow on food and feed matrices and significantly harm the health of humans and farm animals. Among the reported mycotoxigenic fungi, Fusarium graminearum has received wide attention due to its ability to produce multiple mycotoxins, including deoxynivalenol (DON), nivalenol (NIV), and zearalenone (ZEA) under diverse climatic conditions. The present study focused on proving in vitro cytotoxicity of F. graminearum toxins (DON, NIV, and ZEA) in H9c2 cells (Rattus norvegicus heart/myocardi). The effect of F. graminearum toxins (FGTs) on cell viability was studied by MTT, LDH, and live/dead cell assays. The cell viability decreased with increasing the quantity of FGTs and was noticed as dose-dependent. The cell viability assays indicated that DON was significantly more toxic than NIV, and both were considerably more toxic than ZEA. The toxicity mechanism of FGTs was revealed by estimating reactive oxygen species (ROS), mitochondrial membrane potential (MMP), and caspase-3 levels. The effect of FGTs on the generation of ROS was observed by fluorescent probe dichloro-dihydro-fluorescein diacetate (DCFH-DA), and the ROS levels were enhanced on the treatment of FGTs. The effect of FGTs on MMP was revealed by rhodamine 123 staining, and MMP levels were depleted with exposure to FGTs. Furthermore, the expression level of caspase-3 was determined by a caspase-3 detection kit, and its levels were enhanced by exposure to FGTs. The studies conclude that FGTs have a potent cytotoxic effect on H9c2 cells and induce death by oxidative stress-mediated apoptosis.