Study On The Damage Effect Of T-2 Toxin On TM3 Cells

T-2 toxin is a trichothecene mycotoxins,produced under specific conditions by Fusarium,is widely distributed and are the main contaminants of cereals and cereal products.Together with aflatoxin,T-2 toxin is also considered by the World Health Organization（WHO）and the United Nations Food and Agriculture Organization（UN-FAO）as one of the most dangerous naturally occurring food contaminants.T-2 toxin has been reported to be toxic in the reproductive system of the adult male animal.In this study,mouse Leydig cells（TM3）were used as experimental models for molecular and cell biology,and other techniques to study the toxic effects of T-2 toxin.Objectives:To explore the toxic effect of T-2 toxin on TM3 cells,and to demonstrate the molecular mechanism of its toxic effects,and to provide the theoretical basis for the basic research of T-2 toxin.Methods:TM3 cells were treated with different concentrations of T-2 toxin for 24h.Proliferation rate of treated TM3 cells was measured using MTT assay;intracellular calcium(Ca²⁺)was measured using fluo-2 fluorescent probe AM;antioxidant activities of superoxide dismutase（SOD）,catalase（CAT）and glutathione peroxidase（GSH-Px）were measured using assay kits;oxidation level was measured using malondialdehyde（MDA）and reactive oxygen species（ROS）assay kits;mitochondrial function was evaluated thru ATP,and Na⁺-K⁺ATPase and Ca²⁺-Mg²⁺-ATPase activities;apoptotic cells were measured by flow cytometry;mitochondrial membrane potential（MMP）was evaluated using fluorescence probe Rhodamine-123;expression of apoptosis related genes Bax and Bcl-1 was measured using quantitative PCR;lastly,effect of a free radical scavenger,Trolox,was used to analyze apoptosis rate and cell viability.Results:after 24 h treatment with T-2 toxin,TM3 cells showed 1)decreased SOD,CAT and GSH-Px activities,2)decreased mitochondrial membrane potential and level of ATP,and Na⁺-K⁺ATPase and Ca²⁺-Mg²⁺-ATPase activities,3)increased level of MDA and ROS content with increasing T-2 toxin concentration,4)significantly increased expression of Bax/Bcl-2 and percentage of apoptotic cells in relation to dose concentration,and lastly,5)Trolox reduced the apoptosis rate and improved the survival rate of cells.Conclusion:T-2 toxin 1)can damage the structure and functional integrity of TM3cells,thus is not favorable for cell survival,2)can cause oxidative stress which plays a key role in the process of cell injury in TM3 cells,and 3)can induce apoptosis of TM3 cells closely related to mitochondrial damage.