【摘要】 目的:探讨不同浓度脱氧雪腐镰刀菌烯醇(DON)处理,体外原代培养的人正常食管上皮细胞TAP-1、LMP-2表达的变化。方法:采用Western blot和RT-PCR技术从蛋白水平和mRNA水平上分析不同浓度的DON处理24小时,体外培养的人正常食管上皮细胞TAP-1、LMP-2分子表达的变化规律及其量-效关系。结果:不同浓度(50、100、1 000、2 000μg/L)DON处理人食管上皮细胞24小时后,DON各处理组食管上皮细胞TAP-1蛋白的Western blot半定量检测结果分别为0.47±0.22、0.28±0.16、0.24±0.18和0.21±0.19,统计分析表明DON 100、1 000、2 000μg/L处理组与对照组相比差异有统计学意义(P<0.05)。相关分析表明,在0~2000μg/L浓度范围内,随DON处理浓度增高,食管上皮细胞TAP-1蛋白的表达量逐渐降低(r=-0.595,n=4,P<0.01)。RT-PCR结果发现,经不同浓度DON处理食管上皮细胞TAP-1mRNA的相对表达量虽然呈现先升高后降低的趋势,但与对照组相比无统计学差异。DON各处理组LMP-2蛋白的Western blot半定量检测结果分别为2.25±1.03、3.31±1.35、1.90±1.19和2.12±0.22均高于空白对照组(0.91±0.21),其中100μg/L处理组相对表达量最高,差异有统计学意义(P<0.05)。RT-PCR结果,DON 100μg/L处理组的LMP-2 mRNA的相对表达量最高为2.23±0.56,明显高于对照组,差异有统计学意义(P<0.05)。其余处理组差异无统计学意义。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的TAP-1蛋白的表达,但对TAP-1的mRNA未见显著影响。DON(100μg/L)可以促进食管上皮细胞LMP-2蛋白和mRNA表达,其余各浓度未见明显影响。更多还原

【Abstract】 Objective:To investigate the effects of deoxynivalenol(DON) on TAP-1 and LMP-2 expression in human normal esophageal epithelial cells in vitro.Methods:The human normal esophageal epithelial cells were treated with different concentrations of DON for 24 hours,and the expression changes of TAP-1 and LMP-2 protein and mRNA were detected by Western blot and RT-PCR.Results:The protein of TAP-1 was reduced after treated with different concentrations of DON.The values were 0.47±0.22,0.28±0.16,0.24±0.18 and 0.21±0.19 in 50 μg/L,100 μg/L,1 000 μg/L and 2 000 μg/L treatment groups respectively,especially in 100,1 000 and 2 000 μg/L treatment groups,the values were significantly lower compared with control group(P<0.05).There was a dose-response between expression of TAP-1 and concentration of DON(0-2 000 μg/ml)(r=-0.595,n=4,P<0.01).The level of TAP-1 mRNA was increased first and then decreased,but the difference was not significant stastically(P>0.05).The values of LMP-2 protein were 2.25±1.03,3.31±1.35,1.90±1.19 and 2.12±0.22 in 50,100,1 000 and 2 000 μg/L groups respectively,and the value in 100 μg/L group was increased significantly compared with control group(P<0.05).The level of LMP-2 mRNA in 100 μg/L group was increased significantly compared with control group(P<0.05),but not in the other groups(P>0.05).Conclusion:DON could inhibit the expression of TAP-1 in a dose dependent manner in human normal esophageal epithelial cells in protein level,but not in mRNA level.In 100 μg/L group,DON could promote the expression of LMP-2 in protein and mRNA,but not in the other groups.更多还原