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±0.040、0.453±0.031、0.164±0.011）均低于0 mg/L的T-2毒素组（1.021±0.046,P均＜0.05）；在T-2毒素分别为10、20μg/L时,0.1 mg/L硒组的蛋白聚糖酶-1和蛋白聚糖酶-2 mRNA表达均低于0 mg/L硒组（P均＜ 0.05）.结论 T-2毒素通过上调蛋白聚糖的主要代谢酶蛋白聚糖酶-1和蛋白聚糖酶-2达到降解蛋白聚糖的作用,微量元素硒对T-2毒索引起的软骨细胞蛋白聚糖降解有一定保护作用.
Objective 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-2 toxin could induce cartilage matrix degradation through the upregulation of aggrecanases expression and enzyme activities.Trace element selenium has some protective effect on cartilage proteoglycan degradation induced by T-2 toxins.
出处
《中国地方病学杂志》
CAS
CSCD
北大核心
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）
