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

作　　者：罗明秀 马海江 张甜 田嫁 李正正 陈静宏[2] 曹峻岭[2] LUO Mingxiu;MA Haijiang;ZHANG Tian;TIAN Jia;LI Zhengzheng;CHEN Jinghong;CAO Junling(College of Basic Medicine,Ningxia Medical University,Yinchuan 750004,China;School of Public Health,Xi'an Jiaotong University Health Science Center,Key Laboratory of Trace Elements and Endemic Diseases,National Health and Family Planning Commission,Xi'an 710061,China)

机构地区：[1]宁夏医科大学基础医学院,宁夏银川750004 [2]西安交通大学环境与疾病相关基因教育部重点实验室暨卫生部微量元素与地方病重点实验室,陕西西安710061

出　　处：《宁夏医学杂志》2021年第6期481-484,共4页Ningxia Medical Journal

基　　金：宁夏自然科学基金项目(NO.2018AACO3064);大学生创新创业训练计划项目(NO.NXCX2018145);宁夏医科大学基础医学院“西部一”学科师资建设项目(201-3018160)。

摘　　要：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.

关 键 词：TGF-Β 聚集蛋白聚糖 C-28/I2人软骨细胞系 硫酸化修饰 

分 类 号：R599[医药卫生—内科学]