脱氧雪腐镰刀菌烯醇对人食管上皮细胞TAP-1和LMP-2表达的影响

摘要

ObJ3ctive:To investigate 1he effects of deoxynivalenol (DON ) on TAP-L and LMP-2 expressbn in human nomal esophageal epiihelial eels in vitro .M eihods :The hum an nom al esophageal epithelial cells w ere treated w ith different concentratiDns of D ON fbr 24 hours,and 1he expressiDn changes of TAFH. andLMP-2 protein andmRNA were detected by W estem blot and RT-PCR .R esults :The protein of TAM was reduced after treated w ith different concentratiDns of DON .The valieswere 0 .47+0 .22 ,0 .28+0 .16,0 .24+0 .18 and 0 .21+ 0 .19 in 50 f*g/L 400 f*g/t ,1 000 f*g/L and 2 000 jug/t treatn ent groups respectirefy ,especiaLfy in 100 ,1 000 and 2 000 f*g/t treatn ent groups ,1he valieswere significantly bwer compared with control group (P<0 .05) .There was a dose-response beteeen expressbn of TAFH. And concentration of DON (0-2 000 f*g/m 1) ( r= -0 .595 , n= 4, P<0 .01) .The level of TAM m RNA was increased first and Uien decreased ,but Hie difference was not significant stastmlty(P>0 .05).The values of LM P-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 f*g/t groups respectirefy ,and 1he value in 100 f*g/t group was increased significant^ can- paredwith control group (P<0 .05) .The level of LM P-2 mRNA in 100 f*g/t group was increased significantly compared wilh controlgroup ( P<0 .05 ) ,but not in 1he oiher groups( P>0 .05 ) .C onclusion :D 0 N could inhibit 1he expressbn of TA PH. In a dose dependent m anner in human nom al esophageal epithelial cells in protein level,butnot in mRNA level.In 100 f*g/L group ,D0N could promote 1he expressbn of LM P-2 in protein and m RN A ,but not in 1he oiher groups.%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、2 000 μg/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、2 000 μg/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0～2000 μg/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100 μg/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 100 μg/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(100 μg/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.