降钙素基因相关肽对成骨细胞OPN表达的影响

Effect of Calcitonin Gene-related Peptide on OPN Expression in Osteoblasts

目的:探讨外源性降钙素基因相关肽(Calcitonin gene related peptide,CGRP)对大鼠成骨细胞中骨桥蛋白(Osteopontin,OPN)表达的影响。方法:胶原酶消化法培养大鼠原代成骨细胞,碱性磷酸酶染色及茜素红染色对成骨细胞进行鉴定。CCK-8细胞增殖实验筛选CGRP最适作用浓度。将合适浓度的CGRP作用于成骨细胞,qPCR和Western Blot检测OPNmRNA及蛋白的表达。结果:CGRP浓度为10^(-9) mol/L时,大鼠成骨细胞的增殖活性最强。使用10^(-9) mol/L浓度的CGRP作用于成骨细胞,成骨细胞中OPN mRNA及蛋白的表达在第3天明显上调(P<0.05)。结论:CGRP可以促进成骨细胞中OPN的表达。

Objective：To investigate the effects of Calcitonin gene related peptide on OPN expression in primary cultured rat osteoblasts.Methods：Primary cultured rat osteoblasts were obtained by trypsin and collagenase digestion way from calvarial bone of neonate rats and were identified by ALP staining and alizarin red staining.The appropriate concentration of CGRP was selected by CCK-8assay.The expression of OPN were quantified by qPCR and Western blot after cultured with CGRP.Results：The proliferation activity of osteoblasts enhanced when the concentration of CGRP was 10^-9 mol/L.The expression of OPN mRNA increased on the third day（P〈0.05）,and the expression of OPN protein was consistent with the change of the OPN mRNA.Conclusion：CGRPcould promote the proliferation of osteoblasts when the concentration of CGRP was 10^-9 mol/L.CGRP could promote OPN expression in the primary cultured rat osteoblasts.