转化生长因子β对人颈椎关节突关节软骨细胞基质金属蛋白酶13基因表达的调节作用及意义

Regulating Effects of Transforming Growth Factor-β on Gene Expression of MMP-13 mRNA in Human Hyaline Chondrocytes

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.