DON毒素和硒对软骨聚集蛋白聚糖硫酸化修饰的影响

Effects of DON toxin and selenium on sulphate modification of aggrecan

目的研究大骨节病(Kashin-Beck disease,KBD)可疑致病因子脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)和补硒对软骨细胞外基质聚集蛋白聚糖(aggrecan)硫酸化修饰的影响。方法应用体外单层培养C28/I2人软骨细胞,采用MTT法测定不同浓度DON和不同作用时间下的细胞增殖活性,通过Real-time PCR和Western-blot法分别检测加DON毒素或补硒后软骨细胞的aggrecan、PAPS合成酶2(PAPS synthetase 2,PAPSS2)、PAPS转运体1(PAPS transporter 1,PAPST1)、N-乙酰氨基半乳糖胺-4,6-硫酸转移酶[carbohydrate(N-acetylgalactosamine 4-sulfate 6-O)sulfotransferases 15,CHST15]以及芳基硫酸酯酶B(arylsulfatase B,ARSB)mRNA和蛋白的表达水平。结果 DON毒素可抑制C28/I2软骨细胞系的增殖活性,在DON毒素作用下aggrecan、PAPSS2、PAPST1和CHST15的mRNA和蛋白质表达水平降低,ARSB mRNA和蛋白质表达水平升高,加硒可在一定程度上改善这种状况。结论 DON毒素对软骨细胞的增殖有明显的抑制作用,细胞对毒素有一浓度和时间的依赖性;DON毒素可通过改变aggrecan硫酸化修饰相关酶类的表达水平影响软骨蛋白聚糖的硫酸化。

Objective To study the effects of deoxynivalenol （DON）, which is the suspected pathogenic factor of Kashin-Beck disease （KBD）, and selenium supplement on sulphate modification of aggrecan in cartilage extracelluar matrix. Methods C28/I2 human chondrocytes were cultured in monolayer in vitro. Then MTT was used to assay the proliferation of the chondrocytes treated with DON toxin of different concentration and at different action time. Real-time PCR and Western blot methods were used to detect the expressions of aggrecan mRNA and protein, PAPS synthetase 2 （PAPSS2）, PAPS transporter 1 （PAPST1）, carbohydrate （N-acetylgalactosamine 4-sulfate 6-O） sulfotransferases 15 （CHST15） and arylsulfatase B （ARSB）, respectively. Results DON toxin could inhibit the proliferation of C-28/I2 chondrocytes; the mRNA and protein expressions of aggrecan, PAPSS2, PAPST1 and CHST15 were decreased by DON toxin intervention. On the other hand, the mRNA and protein expressions of ARSB were increased, and selenium supplement could improve this situation. Conclusion DON toxin can obviously inhibit the growth of chondrocytes in concentration and action time-dependent manners to some extent. DON toxin affects sulphate modification of cartilage proteoglycan by altering the expression of some related enzymes.