T-2毒素和硒对人软骨蛋白聚糖酶的影响  

Effects of T-2 toxin and selenium on expression of aggrecanase in human chondrocyte

作　　者：于伯泉[1] 曹峻岭[1] 陈静宏[1] 师钟丽[1] 王伟[1] 杨占田[1] 马天有[1] 王世捷[1] 

机构地区：[1]西安交通大学医学院地方病研究所环境与疾病相关性基因教育部重点实验室,710061

出　　处：《中国地方病学杂志》2012年第1期46-50,共5页Chinese Jouranl of Endemiology

基　　金：国家自然科学基金（31070725、30872187）;教育部留学回国人员启动基金（11回国基金01）

摘　　要：目的 研究T-2毒素和硒对人软骨蛋白聚糖代谢的影响.方法 体外培养胎儿软骨细胞,根据施加的实验措施将其分为8组,对照组（T-2毒素：0 mg/L,硒：0 mg/L）、1、10、20μg/L T-2毒素组、加硒（0.1mg/L）组、1、10、20 μg/L T-2毒素加硒（0.1 mg/L）组,每组设3个平行样.在T-2毒素和（或）硒作用5d后,采用免疫蛋白印迹法检测软骨细胞蛋白聚糖的蛋白表达水平,用RT-PCR方法检测软骨细胞蛋白聚糖酶-1和蛋白聚糖酶-2的mRNA表达水平.结果 硒、T-2毒素对蛋白聚糖的蛋白表达量、蛋白聚糖酶-1 mRNA表达水平、蛋白聚糖酶-2的mRNA表达水平有影响（F=0.294、27.71,107.45、362.25,34.68、22.26,P均＜0.05）,硒与T-2毒素存在交互作用（F=79.99、230.76、388.33,P均＜0.05）,表现为拮抗作用.在硒水平为0 mg/L时,1、10、20 μg/L的T-2毒素组蛋白聚糖的蛋白表达（0.278±0.015、0.235±0.029、0.195±0.028）均低于0 mg/L的T-2毒素组（0.472±0.0358,P均＜0.05）；在硒水平为0.1 mg/L时,1、10、20 μg/L的T-2毒素组的蛋白聚糖的蛋白表达（0.399±0.028、0.299±0.020、0.263±0.019）均高于0 mg/L的T-2毒素组（0.197±0.018,P均＜0.05）.在T-2毒素分别为1、10、20μg/L时,0.1 mg/L硒组的蛋白聚糖的蛋白表达均高于0 mg/L硒组（P均＜ 0.05）.在硒水平为0 mg/L时,1、10、20μg/L的T-2毒素组蛋白聚糖酶-1 mRNA表达（0.535±0.033、1.071±0.043、1.454±0.058）均高于0 mg/L的T-2毒素组（0.481±0.023,P均＜0.05）；在硒水平为0.1 mg/L时,1、10、20μg/L的T-2毒素组的蛋白聚糖酶-1 mRNA表达（1.057±0.048、0.555±0.036、0.902±0.045）均高于0 mg/L的T-2毒素组（0.346±0.020,P均＜0.05）.在硒水平为0 mg/L时,1、10、20μg/L的T-2毒素组蛋白聚糖酶-2 mRNA表达（0.596±0.038、0.656±0.033、0.949±0.049）均高于0 mg/L的T-2毒素组（0.387±0.020,P均＜0.05）；在硒水平为0.1 mg/L时,1、10、20μg/L的T-2毒素组的蛋白聚糖酶-2 mRNA表达（0.600±0Objective To determine effects of T-2 toxin and selenium on expression of aggrecanase in human chondrocyte.Methods Human chondrocytes were treated with T-2 toxin（0,1,10,20 μg/L）,and/or sodium selenite（final concentration of selenium 0,0.1 mg/L） for 5 days.Aggrecan expression was determined by Western blotting,aggrecanase-1 and aggrecanase-2 mRNA levels were measured by RT-PCR.Results Selenium and T-2 toxin had effects on both aggrecan protein levels and its aggrecanases （include aggrecanase-1 and aggrecanase-2 ） mRNA levels（F =0.294,27.71 for aggrecan,F =107.45,362.25 for aggrecanase-l,F =34.68,22.26 for aggrecanase-2,respectively,all P 〈 0.05）,and there was interaction between selenium and T-2 toxin on aggrecan,aggrecanase-1 and aggrecanase-2 expression（F =79.99,230.76,388.33,all P 〈 0.05）.Furthermore,selenium presented significant antagonism to T-2 toxin on aggrecan,aggrecanase-1 and aggrecanase-2 expression.Aggrecan expression levels（0.278 ± 0.015,0.235 ± 0.029,0.195 ± 0.028,0.399 ± 0.028,0.299 ± 0.020,0.263 ±0.019） induced by both 1,10,20 μg/L T-2 toxin and 0,0.1 mg/L selenium were significantly decreased than the levels（0.472 ± 0.0358,0.197 ± 0.018,all P 〈 0.05） in control group（0 mg/L toxin）.Selenium partially blocked the effects induced by 1,10,and 20 μg/L T-2 toxin（all P〈 0.05）.One,10,20 μg/L T-2 toxin and 0,0.1 mg/L selenium increased both aggrecanase-1 mRNA levels（0.535 ± 0.033,1.071 ± 0.043,1.454 ± 0.058,1.057 ±0.048,0.555 ± 0.036,0.902 ± 0.045） and aggrecanase-2 mRNA levels（0.596 ± 0.038,0.656 ± 0.033,0.949 ±0.049,0.600 ± 0.040,0.453 ± 0.031,0.164 ± 0.011）,when compared with control（0.481 ± 0.023,0.346 ±0.020 for aggrecanase-1 and 0.387 ± 0.020,1.021 ± 0.046 for aggrecanase-2,respectively,all P 〈 0.05）.Selenium partially blocked 10,20 μg/L T-2 toxins induced upregulation of aggrecanase-1 （all P 〈 0.05） and aggrecanase-2 （all P 〈 0.05 ）.Conclusions These data suggest a possible molecular mechanism that T-

关 键 词：硒 软骨细胞 大骨节病 蛋白聚糖酶 T-2毒素 

分 类 号：R684.1[医药卫生—骨科学]