摘要
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.
出处
《中国修复重建外科杂志》
CAS
CSCD
北大核心
2005年第8期607-610,共4页
Chinese Journal of Reparative and Reconstructive Surgery
基金
国家教育部骨干教师资助计划项目(2000-7)~~