甲状旁腺素促进小鼠软骨细胞成软骨和抑制其终末期分化的实验研究

目的探讨甲状旁腺素(PTH)对小鼠软骨细胞成软骨性的促进作用和终末期分化的抑制作用。方法分离和培养新生小鼠胸骨软骨细胞,经PTH处理,倒置显微镜观察细胞形态的变化;Alcian蓝染色和碱性磷酸酶(ALP)染色方法检测软骨细胞蛋白多糖和ALP的分泌;RT-PCT法和Western blot方法检测细胞内成软骨因子和病理性肥大分化因子基因和蛋白的表达。结果新生小鼠胸骨软骨细胞具有自发成熟分化的特征,与对照组相比,经PTH处理的细胞更接近于软骨细胞形态;PTH明显提高软骨细胞Alcian蓝染色的强度,降低ALP染色的强度;PTH显著提高细胞内Sox9和Aggrecan基因和蛋白的表达,明显降低ALP和Runx2基因和蛋白的表达。结论 PTH具有促进小鼠软骨细胞成软骨和抑制其终末期分化的作用。

Investigation of the chondrogenic and anti-terminal maturation effects of parathyroid hormone on mouse chondrocytes

Objective To investigate the chondrogenic effect of parathyroid hormone （PTH） on mouse sternal chondrocytes （MSCs） and the inhibition of terminal differentiation of MSCs by PTH. Methods Mouse sternal chondrocytes were isolated and cultured. The cells were treated with PTH. The morphology of ceils was observed by microscopy. Alcian blue staining and alkaline phosphatase （ALP） staining were performed to detect the polyproteoglycan and ALP protein. The chondrogenic marker genes and hypertrophic marker genes mRNA and protein expressions were detected by real-time RT-PCR and Westernblot. Results MSCs underwent spontaneous maturation. The cells treated with PTH have more phenotypes of chondrocytes compared to controls. The Alcian blue staining density was enhanced by PTH treatment, while the ALP staining density was decreased with PTH treatment. In addition, PTH increased Sox9 and Aggrecan mRNA and protein expression dramatically and decreased ALP and Runx2 mRNA and protein expression dramatically. Conclusion PTH has chondrogenic effect on MSCs and inhibits terminal differentiation of MSCs.