转化生长因子β对人透明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 MATRIX METALLOPROTEINASE 13 mRNA IN HUMAN HYALINE CHONDROCYTES

目的探讨转化生长因子β(transforminggrowthfactorβ,TGF-β)对人透明软骨细胞基质金属蛋白酶13(matrixmetalloproteinase13,MMP-13)基因表达的作用,了解骨关节炎软骨损伤的机制。方法将体外培养的人软骨细胞随机分为7组:第1组为正常对照组,第2、3、4组分别加入1、10和100ng/mlTGF-β,第5、6、7组分别加入前3种不同浓度TGF-β后再加入10ng/ml白介素1β(interleukin1β,IL-1β)作为联合作用组。每组均作用12h,应用逆转录-聚合酶链反应(reversetranscription-polymerasechainreaction,RT-PCR)及实时荧光定量PCR,检测不同因子作用下传代培养人透明软骨细胞中MMP-13mRNA的含量。结果第1组透明软骨细胞仅见MMP-13mRNA扩增产物;第2、3和4组,MMP-13mRNA表达逐渐增强;第5、6、7组,随着TGF-β浓度的升高,MMP-13mRNA表达逐渐降低,且各组之间差异有统计学意义(P<0.05)。结论MMP-13在骨关节炎的发生与发展中有一定的作用,TGF-β可调节软骨细胞MMP-13mRNA基因的表达。

Objective To investigate the role of transforming growth factor 13（TGF-β）in the regulation of the gene expression of matrix metalloproteinase 13 （MMP-13）in the human hyaline chondrocytes. Methods The human hyaline chondrocytes harvested enzymatically and cultured in DMEM supplemented with 20% fetus calf serum were divided into 7 groups. Group 1 was used as a contol, and 1 ng/ml TGF-β（group 2）, 10 ng/ml TGF-β（group 3）, 100 ng/ml TGF-β（group 4）. 1 ng/ml TGF-β＋10 ng/ml IL-1β（group 5）, 10 ng/ml TGF-β＋10 ng/ml IL-1β（group 6）, and 100 ng/ml TGF-β＋ 10 ng/ml IL-1β（group 7） were given for 12-hour coculture. The MMP-13 mRNA levels of passaged human hyaline chondrocytes were assessed by reverse transcription-polymerase chain reaction（RT-PCR） and real-time fluorescent quantitative PCR. Results TGF-β can increase the MMP-13 mRNA level respectively in the passaged hyaline chondrocytes. In the multi-factor treated groups, TGF-β can decrease the MMP-13 mRNA level respectively and there was significant difference between groups （P〈0. 05）. The level of MMP-13 mRNA expression had significant coherence with the dosage of TGF-β. Conclusion The above results show that human chondrocytes express MMP-β mRNA. MMP-13 could cause a dose dependent stimulation on MMP-13 gene expression in human chondrocytes and have a potent effect of antagonizing IL-1β in osteoarthritis. TGF-β may play a crucial role in the occurrence and development of osteoarthritis through regulating MMP-13.