45S5对体外培养的鼠鼻中隔软骨细胞骨化基因表达的影响

目的探讨45S5对体外培养的鼠鼻中隔软骨细胞成骨分化的目的基因活化诱导作用。方法利用体外细胞培养技术建立孕21 d SD大鼠鼻中隔软骨细胞培养体系,根据培养基的不同分为实验组(培养基中放入45S5 1 mg/mL)和空白对照组。第2代软骨细胞培养至5、9、15、20 d取材,RT-PCR检测Run X2和CollagenX的mRNA的表达;8、12、15、18 d取材分光比色检测碱性磷酸酶(ALP)。结果45S5诱导培养下第9天的Run X2及第20天的type X collagen mRNA的表达明显增强,第10、15天ALP的分泌明显高于对照组(P<0.05)。结论45S5可促进体外培养的鼠鼻中隔软骨细胞增值及向成骨细胞分化。

Regulating effects on gene expressions of endochondral ossification in rat nasal chondrocyte cultures by 45S5

Objective To explore the modulation of endochondral ossification in rat nasal chondrocyte cultures by 45S5 in vitro. Methods Chondrocytes were enzymatically isolated from the nasal septum of 21 day-old foetal Sprague Dawley rats and cultured up to 8、12、15、18 days in the presence of a melt-derived 45S5 and control group. The specific activity of alkaline phosphatase were determined in a by atomic absorption spectrometry. Cultured up to 5、9、15、20 days, The RunX2 andCollagenX mRNA levels of chondrocyte cultured were assessed by real time RT-PCR. Results The specific activity of alkaline phosphatase was significant higher in 45S5 cultures on day12 and 15 than control group. Runx2 expression was higher in experiment cultures on day 9. type X collagen mRNA seemed to be up-regulated in 45S5 cultures on day 20. Conclusions This study shows 45S5 has the ability to modulate the growth of chondrocytes and endochondral ossification.