【摘要】 目的探讨不同剂量维生素 C(vitamin C,VC)对脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)诱导的体外培养人外周血单个核细胞(human peripheral blood mononuclear cells,HPBMC)HLA-Ⅰ表达抑制作用的影响。方法采用流式细胞术(flow cytometry,FCM)、免疫细胞化学和蛋白印迹(Western blotting)方法研究了不同剂量(25μmol/L 和100μmol/L)VC 预处理对 DON 诱导的 HPBMCHLA-Ⅰ表达抑制作用的影响。结果 FCM 定量检测结果表明,DON 处理组 HPBMC HLA-Ⅰ的表达与正常对照组相比明显降低(FI 值0.88±0.02比1.00±0.03,P<0.05)。25μmol/L 和100μmol/LVC 预处理+DON 组 HPBMC HLA-Ⅰ表达的 FI 值分别为1.15±0.06和1.10±0.02,均明显高于DON 处理组(0.88±0.02,P<0.05)。同时,25μmol/L 和100μmol/L VC 单独处理组 HPBMC HLA-Ⅰ的表达量也均明显高于对照组(FI 值1.28±0.03和1.25±0.05比1.00±0.03),表明 VC 可在一定程度上促进体外培养的 HPBMC HLA-Ⅰ表达(P<0.05)。免疫细胞化学结果表明,与 DON 处理组相比,两种不同剂量 VC 预处理+DON 组 HLA-Ⅰ表达阳性的细胞百分率比 DON 处理组明显增高(70.10%±6.90%和64.50%±5.50%比42.20%±4.30%,P<0.05)。蛋白印迹同样证实了以上结果。结论 VC 可一定程度的拮抗 DON 对体外培养的 HPBMC HLA-Ⅰ表达的抑制作用。更多还原

【Abstract】 Objective To explore the putative effects of Vitamin C (Vit C) on inhibition of human leucocyte antigen class Ⅰ (HLA-Ⅰ)expression of human peripheral blood mononuclear cells (HPBMCs) induced by deoxynivalenol (DON) in vitro.Methods The effects of Vit C on the changes of HLA-Ⅰ expression of HPBMCs induced by DON/n vitro were evaluated with cell culture,flow cytometry ( FCM ), Western blotting and immunocytochemieal methods.Results FCM analysis showed that HLA-Ⅰ expression of HPBMCs in DON treated cells was significantly lower than that in controls ( FI 0.88±0.02 vs 1.00± 0.03,P<0.05 ).As compared with DON group,the HLA-Ⅰ expressions of HPBMCs in the two Vit C (25 μmol/L and 100 μmol/L) pretreatment groups were all significantly increased (1.15±0.06 and 1.10± 0.02 vs 0.88±0.02,P<0.05).Exposure to different dosage of Vit C alone could dramatically increase the expression of HLA-I of HPBMCs in vitro as compared with that in the normal control ( FI for 25 μmol/L and 100 μmol/L Vit C treatment group was 1.28±0.03 and 1.25±0.05 respectively,P<0.05). Immunocytoehemical results showed that the percentages of HLA-Ⅰ positive expression of HPBMCs in Vit C pretreatment groups at different dosages were significantly higher than those in DON group (70.10± 6.90) %,(64.50±5.50)% vs ( 42.20±4.30 ) %,P<0.05.Western blotting confirmed the results of FCM and immunocytochemistry.Conclusion Vitamin C pretreatment at different dosages could reverse at some extent the inhibitive effects of DON on HLA-Ⅰ expression of HPBMCs.更多还原