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目的探讨脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)对人正常食管上皮细胞(human leukocyteantigen-Ⅰ,HLA-Ⅰ)表达的影响。方法采用流式细胞技术(flow cytometry,FCM)、免疫细胞化学染色(immunohistochemistry,IHC)以及反转录聚合酶链反应(reversetranscliption-polymerase chain reaction,RT-PCR)技术,从蛋白水平和mRNA水平上分析不同浓度DON处理24h后,体外培养的人正常食管上皮细胞HLA-Ⅰ表达的变化及剂量-效应关系。结果 IHC检测结果,不同浓度(50、100、1 000、2 000μg/L)DON处理24 h后,各处理组HLA-Ⅰ蛋白阳性细胞百分数均低于空白对照组,其中100、1000和2 000μg/L处理组差异有统计学意义(P<0.05);相关分析表明在0～2 000μg/L范围内,DON浓度与HLA-Ⅰ蛋白阳性细胞百分数之间存在明显的剂量-效应关系(r=-0.756,P<0.05)。FCM检测结果表明,DON在0～2 000μg/L范围内,随着浓度增加食管上皮细胞HLA-Ⅰ蛋白表达的荧光指数逐渐降低,两者之间存在明显的剂量-效应关系(r=-0.634,P<0.05)。RT-PCR结果显示,各处理组HLA-a mRNA的表达量与空白对照组相比,差异无统计学意义(P>0.05)。结论 DON可剂量依赖地抑制体外培养的人正常食管上皮细胞HLA-Ⅰ蛋白的表达,但是对于mRNA表达无明显影响。
Objective To investigate the effect of deoxynivalenol (DON) on the expression of human leukocyte antigen-Ⅰ (HLA-Ⅰ) in human normal esophageal epithelial cells. Methods Flow cytometry (FCM), immunocytochemistry (IHC) and reverse transcriptase-polymerase chain reaction (RT-PCR) The changes of HLA-Ⅰ expression in normal human esophageal epithelial cells incubated with different concentrations of DON for 24 hours and the dose-response relationship were analyzed. Results IHC results showed that the percentage of HLA-Ⅰ positive cells in each treatment group was lower than that of the blank control group after treated with different concentrations of DON (50, 100, 1 000 and 2 000 μg / L) for 24 h, 100, 1000 and 2 000 μg (P <0.05). Correlation analysis showed that there was a significant dose-response relationship between DON concentration and the percentage of HLA-Ⅰ positive cells in the range of 0-2 000 μg / L (r = -0.756, P <0.05). The results of FCM showed that the fluorescence index of HLA-Ⅰ protein in esophageal epithelial cells decreased gradually with the concentration of DON in the range of 0-2 000 μg / L, and there was a significant dose-response relationship between the two (r = -0.634 , P <0.05). The results of RT-PCR showed that there was no significant difference in the expression of HLA-a mRNA between the treatment groups and the blank control group (P> 0.05). Conclusion DON inhibits the expression of HLA-Ⅰ protein in normal human esophageal epithelial cells in a dose-dependent manner, but it has no significant effect on mRNA expression.