Research On Oxidative Injury And Apoptosis On Combined Effects Of Deoxynivalenol And Zearalenone On Porcine Splenic Lymphocytes In Vitro Culture

Deoxynivalenol (DON) and zearalenone (ZEA) are the two most common mycotoxins in feed. DON, ZEA not only reduces the nutritional value, resulting in animal growth performance, reproductive performance, but also damage the immune system. In recent years, their immune toxicity increasing attention. In this study, by CCK-8 test, the IC50 of DON and ZEA to porcine splenic lymphocytes in vitro at 48 h were 1.48±0.20?g/mL and 2.03±0.31?g/mL, individually. So, it showed that DON or ZEA affected cell proliferation activity. Based on the above IC50, we designed DON individual exposure dose level of 0.06, 0.3,1.5,7.5?g/mL, ZEA individual exposure dose level of 0.08,0.4,2, 10?g/mL, DON and ZEA combined exposure dose of 0.06?g/mL and 0.08?g/mL,0.3?g/mL and 0.4?g/mL, 1.5?g/mL and 2?g/mL, a separate control group. DON and/or ZEA exposure porcine splenic lymphocytes after 48h, detection antioxidant function, apoptosis, apoptosis-related genes p53, Bcl-2, Bax, Caspase-3 Caspase-8 mRNA expression and the culture supernatant and apoptosis-related gene protein content, research DON. ZEA alone or in combination with exposure to porcine spleen lymphocyte apoptosis and oxidative damage effects and mechanisms, in order to further clarify DON, ZEA and their combined immunodeficiency toxicity mechanism provides a theoretical basis It is also a useful complement to previous studies.The results showed that:(1) Compared with the control group, except for DON, ZEA individual exposure the lowest dose group of individual indicators, the exposed group SOD, CAT, GSH-Px activity and GSH levels were significantly reduced (P<0.05 or P< 0.01). MDA content increased significantly (P<0.01), and has a dose dependent manner; the role of joint exposure group effect five indicators were higher than the corresponding single-dose exposure group, DON+ZEA displayed sub-additive effects. (2) Each exposure group apoptosis rate was significantly higher than the control (P<0.01), with the toxin dose increased in a dose-dependent effect, the combined group apoptosis rate was significantly higher corresponding individual exposure group (P<0.01), showed significant interaction effects were sub-additive effects. (3) Compared with the control group, except for DON, ZEA individual exposure the lowest dose group of individual indicators, the exposed group p53, Bax, Caspase-3 and Caspase-8 content and mRNA expression were significantly increased water (P<0.05 or P<0.01), while Bcl-2 content and mRNA expression levels were significantly decreased (P<0.05 or P<0.01), the change in a dose dependent manner; Except for the D1Z1 group, the impact of combined exposure group above indexes were greater than the corresponding single dose exposure group, DON+ZEA displayed sub-additive effects.Conclusion:(1) Fusarium mycotoxins DON and/or ZEA can cause intracellular antioxidant enzyme activity decreased, increased oxidation products resulting from splenic lymphocytes oxidative damage, the degree of oxidative damage consistent with apoptosis,oxidative damage is one of the mechanisms of apoptosis death. (2) DON and/or ZEA increased Bcl-2 family, Bax gene expression levels of genes, Bcl-2 expression levels decrease, increased p53, Caspase-3 and Caspase-8 gene expression, indicating that DON and ZEA inducted apoptosis by Caspase and p53 pathway and apoptosis were regulated by Bcl-2 family of proteins. (3) DON+ZEA displayed sub-additive effects on oxidative damage and apoptosis on porcine splenic lymphocytes.