摘要
目的探讨转化生长因子β(TGF-β)对人的颈椎关节突关节透明软骨细胞基质金属蛋白酶13(MMP-13)基因表达的作用,旨在阐明颈椎退行性变的相关发生机理。方法应用逆转录方法PCR及实时荧光定量方法,检测不同浓度TGF-β作用传代培养人的透明软骨细胞MMP-13 mRNA的含量。另外3种不同浓度分别与10ng/mlIL-1β组成联合作用组,共计6个实验组及1个正常对照组。结果正常对照组中透明软骨细胞仅见MMP-13 mRNA扩增产物,实验组TGF-β1、10和100ng/ml作用12h后,MMP-13 mRNA表达逐渐增强;而联合作用组中,随着TGF-β1浓度的升高,MMP-13 mRNA表达逐渐降低,并且各组之间存在明显的差异(P<0.05)。结论TGF-β可按剂量依赖方式调节颈椎关节突关节软骨细胞MMP-13 mRNA的表达。
Objective To investigate the role of transforming growth factor β (TGF - β) in the regulation of the gene expression of matrix metaUoproteinase- 13 (MMP - 13) in the human hyaline chondrocytes of zygapophysial joint of the cervical vertebrae. Methods In the experiment, the human hyaline chondroeytes were harvested enzymatically and cultured in DMEM supplemented with 20% fetus calf serum, and given respectively 1 ng/ml TGF -β, 10 ng/ml TGF- β, 100 ng/ml TGF - β, 1 ng/ml TGF -β + 10 ng/ml IL - 1β, 10 ng/ml TGF-β+ 10 ng/ml IL- 1β, 100 ng/ml TGF- β+ 10 ng/ml IL- 1β, and cultured for 12 hours. The passaged monolayer cell cultures of human hyaline chondrocyte MMP- 13 mRNA level were assessed by the reverse transcriptase polymerase chain reaction (RT - PCR) and the fluorescent quantitative PCR (FQ - PCR). Results TGF - β increased the MMP - 13 mRNA level respectively in passaged cultures of hyaline chondrocyte. In the multi - factor treated groups, TGF - 13 decreased the MMP - 13 mRNA level respectively with the significant difference (P 〈 0.05 ). The level of MMP- 13 mRNA expression had significant correlativity with the dosage of TGF-β1. Conclusion TGF-β1 could cause a dose- dependent stimulation on MMP - 13 gene expression in human chondrocyte and has a potent effect antagonizing IL- 1 in the cervical degenerative disease.
出处
《中国骨与关节损伤杂志》
2007年第3期204-206,共3页
Chinese Journal of Bone and Joint Injury
关键词
基质金属蛋白酶
基因表达
转化生长因子
软骨细胞
Matrix metalloproteinase
Gene expression
Transforming growth factor
Chondrocytes