Protective Effect Of Betulinic Acid On T-2 Toxin-induced Oxidative Damage In Brain Tissue Of Mice

T-2 toxin is a mycotoxin which produced mainly by a variety of Fusarium,and it contaminates wheat,barley,corn and other food crops and their products.Animals whom consumed the feed contaminated by T-2 toxin or long-term exposure to it may induce oxidative stress in the body,and lead to the damage of immune system,reproductive system,circulatory system and cognitive function.Consequently it causes great damage to the livestock and poultry industry.Betulinic acid(BA)is a plant-derived pentacyclic triterpenoid that exhibits various biological activities such as immunomodulation,anti-oxidative stress and anti-inflammation.Notably,BA may have promising clinical applications in the animal farming as an immunomodulator and antioxidant.It was found that BA alleviated the occurrence of brain-related diseases by improving the antioxidant capacity of brain.However,the preventive effect of BA on T-2 toxin induced brain oxidative injury has not been reported in literature.Objective:In this study,a model to induce brain oxidative stress was established in mice by injecting T-2 toxin.The objective was to explore the preventive effect of BA on T-2toxin-induced brain oxidative stress in mice which may help to form the theoretical basis for the establishment of new methods to alleviate brain oxidative damage,and to improve brain health.Methods:Total of 60 healthy male mice were randomly divided into 6 groups,namely,the control group,T-2 toxin group,low,medium and high doses of BA(0.25 mg / kg,0.5 mg / kg,1 mg/ kg)+ T-2 groups,V-E + T-2 group.BA and V-E were suspended in 1% soluble starch,and the control group and T-2 group were given the same amount of 1% soluble starch once a day and continuously administered for 14 days.Animals were received T-2 toxin intraperitoneally at the dose of 4 mg/kg b.w.to induce oxidative damage.After fasting for15 h(free drinking water),mice were executed to collect blood and brain.The levels of acetylcholine(ACh),malondialdehyde(MDA),glutathione(GSH),catalase(CAT),superoxide,dismutase(SOD),and glutathione peroxidase(GSH-Px)in the brain were determined by commercial assay kits.The levels of dopamine(DA),5-hydroxytryptamine(5-HT),were detected by ELISA.The morphological changes and ultrastructure of brain were observed by H&E stain and transmission electron microscopy,respectively.Cell apoptosis in brain was detected by TUNEL,and m RNA expression of cytokines such as interleukin-1 beta(IL-1β),IL-6,IL-10 and tumor necrosis factor-α(TNF-α)in brain were determined using RT-PCR.Results:(1)BA pretreatment alleviated the decrease of SOD,CAT and GSH-Px activities in brain caused by T-2 toxin,increased the content of GSH,and decreased the content of MDA,(2)BA pretreatment reduced the increase of ROS level in brain tissue caused by T-2 toxin;(3)BA pretreatment alleviated the reduction of ACh,DA and 5-HT levels and other neurotransmitters in brain;(4)BA reduced brain bleeding and congestion,improved the mitochondrial morphology,enriched the number of organelles,and inhibited cell apoptosis in brain;(5)BA pretreatment decreased m RNA expressions of IL-1β,IL-6 and TNF-α,and increased m RNA expression of anti-inflammatory cytokine such as IL-10.Conclusion:BA had a protective effect on T-2 toxin-induced brain oxidative damage in mice by reducing oxidative stress,inhibiting the secretion of inflammatory cytokines,and improving cognitive function.