TGF-β对C-28/I2人软骨细胞聚集蛋白聚糖硫酸化修饰的影响

The effect of TGF-β on sulphate modification of aggrecan in C-28/I2 human chondrocytes

目的观察TGF-β降低对C-28/I2人软骨细胞外基质聚集蛋白聚糖(aggrecan)硫酸化修饰的影响。方法设计构建3个不同靶点人源TGF-β1干扰慢病毒载体,荧光鉴定病毒包装成功后将其感染C-28/I2人软骨细胞系,感染72 h后提取细胞总蛋白,用Western-Blot法分别检测TGF-β1以及aggrecan硫酸化修饰相关酶蛋白因子PAPSS2、PAPST1、CHST15蛋白的相对表达量。结果荧光检测结果表明,TGF-β1干扰慢病毒载体构建成功。TGF-β1慢病毒感染C-28/I2人软骨细胞后与阴性对照病毒组相比,TGF-β1、PAPSS2、PAPST1和CHST15的蛋白表达均有明显降低,其中TGF-β1和PAPSS2的蛋白表达降低为最显著,差异具有统计学意义(P<0.05)。结论TGF-β信号可能是通过调控下游相关酶蛋白因子的表达来对软骨细胞aggrecan硫酸化的修饰产生一定的影响。

Objective To observe the effect of TGF-β on the sulphate modification of aggrecan in C-28/I2 human chondrocyte extracellular matrix.Methods Three human TGF-β1 interfering lentiviral vectors with different targets were designed and constructed.The total protein of the infected-cells was extracted after the C-28/I2 human cell line was infected with TGF-β1 lentiviral vectors for 72h.The relative expression levels of TGF-β1,PAPSS2,PAPST1 and CHST15 were detected by Western-blotting.Results TGF-β1 interfered with lentiviral vector was successfully constructed.Compared with the negative control virus group,the protein expressions of TGF-β1,PAPSS2,PAPST1 and CHST15 were decreased in C-28/I2 human chondrocytes after TGF-β1 infected,and the expression of TGF-β1 and PAPSS2 were significantly decreased(P<0.05).Conclusion TGF-βsignal may play a role in the sulphate modification of aggrecan in chondrocytes by regulating the expression of downstream enzyme.