脱氧雪腐镰刀菌烯醇抑制人食管上皮细胞HLA-Ⅰ分子蛋白表达

THE EXPRESSION OF HLA-I PROTEIN WAS REDUCED BY DEOXYNIVALENOL IN HUMAN ESOPHAGEAL EPITHELIAL CELLS IN VITRO

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 effects of deoxynivalenol （DON） on the expression of human leukocyte antigen - I （HLA -I ） in human normal esophageal epithelial cells in vitro. Methods The expression of HLA - I mRNA and protein were detected with reverse transcription reverse transcription - polymerase chain reaction （ RT - PCR ）, immunohistochemistry （ IHC ） and flow cytometry （FCM） in human normal esophageal epithelial ceils treated by DON for 24 hours. Results The results of IHC showed that the percentage of cells with HLA - I expression in treatment groups were lower that in control group, especially in 100,1 000 and 2 0001xg/L groups （P 〈0.05 ）. The correlation analysis showed that there was a significant dose -response relationship between DON and percentage of positive cells （r = - 0.756, P 〈 0.05）. The results of FCM suggested that the FI of HLA - I in treatment groups were lower than that in control group, there was a significant dose -response relationship between DON and FI （ 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 treatment groups and control group （ P 〉 0.05 ）.Conclusion DON could inhibit the expression of HLA - Iprotein, not mRNA in a dose - response manner in human normal esophageal epithelial cells in vitro, but had no obvious effect on mRNA.