碱性成纤维细胞生长因子及其受体在体外培养软骨细胞中的表达

目的 探讨软骨细胞在体外传代培养过程中碱性成纤维细胞生长因子(bFGF)及其受体(FGFR)表达的变化趋势，以及它对软骨细胞增殖和细胞外基质(ECM)合成的影响。方法 采用猪耳软骨细胞在体外进行扩增传代，收集第1～6代的细胞样品行RT-PCR和Western blotting检测，从mRNA和蛋白水平来反映bFGF和FGFRs的表达。另取第1～6代软骨细胞，以含10ng／mL bFGF的培养液进行培养，比较其与空白对照组细胞在增殖速率和ECM合成等方面的差异。结果 bFGF、FGFR1和FGFR2在第1～3代软骨细胞中表达减少的趋势较为平缓；第4、5代时，有大幅的向下跃迁；在第6代时仅微量表达或消失。这一现象与体外培养软骨细胞增殖速率的下降相平行，第4代及以后代次细胞对bFGF刺激无反应。结论 bFGF和FGFRs表达的减弱与软骨细胞在体外培养过程中的老化进展可能有密切的关系。

Expression of basic fibroblast growth factor and its receptors in porcine chondrocytes cultured in vitro

Objective To determine the relationship between changes in bFGF and FGFR expression and cell viability in in vitro cultural chondrocytes of different passages. Methods Porcine Auricle Chondrocytes were cultured in vitro and harvested from every passage for RT-PCR and Western blotting analysis. Additionally, passage 1-6 chondrocytes were cultured with 10 ng/ml bFGF to investigate the effect of bFGF slowly from passage 1 to passage 3, but decreased dramatically during passage 4 and to them. Results Expression of bFGF and FGFRs in chondrocytes decreased in passage 5 and almost lost the expression in passage 6. The proliferation of chondrocytes cultured in vitro also exhibited a similar pattern. Furthermore, the effect of bFGF on chondrocytes was only observed in passage 1-4. Conclusion Decline of bFGF and FGFR expression may play a key role in the senescence of chondrocytes during their long term culture in vitro.