Research On Sodium Selenite Inhibition Porcine Splenic Lymphocytes Injury Induced By Deoxynivalenol In Vitro Culture

In this study, the effects of deoxynivalenol（DON） on the oxidative damage and sodium selenite （Na₂SeO₃） inhibition of oxidative damage induced by DON of porcine spleen lymphocytes were assessed in vitro after adding of DON and Na₂SeO₃ alone or in combination.Methods:（1） The porcine spleen lymphocytes were cultured in different concentrations of DON （0.0625 μg/mL,0.125 μg/mL,0.25 μg/mL,0.5 μg/mL,1 μg/mL,2 μg/mL,4 μg/mL,8 μg/mL,16 μg/mL,32 μg/mL） and Na₂SeO₃（0.25 μmol/L,0.5 μmol/L,1 μmol/L,2 μmol/L,4 μmol/L,8 μmol/L,16 μmol/L） respectively during 48 h, the cells viability were detected via CCK-8 test and the IC50 of DON and the optimum concentration of Na₂SeO₃ to porcine spleen lymphocytes were calculated respectively.（2） The experiment included D1:（DON 103 ng/mL）, D2 （DON 206 ng/mL） D3 （DON 412 ng/mL）, D4 （DON 824ng/mL）, S （Na₂SeO₃2 μmol/L）, SD1 （DON 103 ng/mL+Na₂SeO₃ 2 μmol/L）, SD2 （DON 206 ng/mL+Na₂SeO₃2 μmol/L） SD3 （DON 412 ng/mL+Na₂SeO₃2 μmol/L）, SD4 （DON 824 ng/mL+Na₂SeO₃2 μmol/L） and blank control group.The porcine spleen lymphocytes were cultured in vitro in 6,12,24 and 48 h. The cells and cells culture supernatants were collected respectively to detect the cells viability, cells supernatant LDH activity, antioxidant enzymes （GPx, SOD, CAT） activities, cellular total antioxidant capacity （T-AOC） and the ability to inhibit the hydroxyl radicals changes in cells, and the levels of MDA, GSH and H2O2.Results:（1） After the porcine spleen lymphocytes were cultured 48 h in vitro, compared with the control group, the groups except the low concentration group （0.0625 μg/mL） suppressed cells viability significantly （P<0.01）. The IC50 of DON to porcine spleen lymphocytes in vitro at 48 h was 0.82±0.35 μg/mL.0.25-4 μmol/L Na₂SeO₃ significantly improved cells viability of porcine spleen lymphocytes （P<0.01）, while Na₂SeO₃ concentration increased to 16 μmol/L the cells viability were significantly decreased （P<0.01）. The optimal nutrition concentration of Na₂SeO₃ to porcine spleen lymphocytes was 2 μmol/L.（2）The porcine spleen lymphocytes cultured with DON alone for 6,12,24 and 48 h, compared with the control group, except individual indicators of group D4 （103 ng/mL）, cells supernatant LDH activity, the levels of MDA and H2O2 of the rest groups were significantly increased respectively（P<0.05 or P<0.01）, but the cells viability, the levels of GSH, antioxidant enzymes activities （GPx, SOD, CAT）, T-AOC and the capacities of inhibition of hydroxyl radicals of the rest groups were significantly decreased respectively （P<0.05 or P<0.01）.（3）The porcine spleen lymphocytes cultured with Na₂SeO₃ alone for 6,12,24 and 48 h, compared with the control group, the cells viability, cells GPx activity（12,24 and 48 h）, T-AOC and the capacity of inhibition of hydroxyl radicals were significantly increased （P<0.05 or P<0.01）. The levels of H2O2 （12 h and 24 h） and MDA （24 h and 48 h） were significantly decreased （P<0.05 or P<0.01）.（4）The porcine spleen lymphocytes cultured with DON and Na₂SeO₃ in combination for 6,12,24 and 48 h. Throughout the experiment period, compared with the corresponding DON groups, the cells viability, the activities of GPx, SOD and CAT, T-AOC and the capacities of inbibition of hydroxyl radical were increased, the levels of MDA and H2O2 and LDH activities of cells supernatant were decreased. At 24 h, compared with the corresponding DON groups, the cells viability（SD3 and SD4 groups）, the levels of GSH, the activities of GPx（except SD1） and CAT（SD3 and SD4 groups）, T-AOC, and the capacity of inbibition of hydroxyl radicals（except SD3 group） were significantly increased （P<0.05 or P<0.01）, while the LDH activity of cells supernatant and the levels of MDA and H2O2 were significantly decreased respectively（P<0.05 or P<0.01）.Conclusions:DON exposure decreased antioxidant function, porcine spleen lymphocytes viability decreased caused by DON.0.25-4 μmol/LNa₂SeO₃ improved the survival rate of lymphocytes, while at 16 μmol/L it produced toxic effects. In this experiment,2 μmol/L of Na₂SeO₃ could inbibite the oxidative damage of porcine spleen lymphocytes induced by DON, and the best time of Na₂SeO₃ to inhibiting oxidative damage was 24...