中文摘要：

目的：探讨外源性降钙素基因相关肽（calcitonin gene-related peptide,CGRP）对大鼠颅盖骨来源成骨细胞RUNX2因子表达水平的影响,进一步了解CGRP促进成骨的作用机制。方法：用酶消化法培养原代大鼠成骨细胞并鉴定;MTT法筛选促进成骨细胞增殖的有效浓度,将含有不同浓度CGRP的条件培养基加入大鼠成骨细胞培养体系中,药物作用48 h后,用半定量RT-PCR和Western blotting方法检测大鼠成骨细胞转录因子RUNX2在mRNA和蛋白水平的表达。结果：当CGRP浓度在10-8～10-6mol/L范围时,对成骨细胞增殖有明显促进作用,增殖率分别为71.9%,142.1%,321.0%,P〈0.05;浓度为10-7和10-6mol/L的CGRP作用于成骨细胞48 h后,转录因子RUNX2在mRNA水平的表达量明显上调,分别增强（46.2±11.2）%和（58.6±14.0）%,P〈0.05;转录因子RUNX2的蛋白表达变化趋势与其mRNA的表达变化趋势基本一致。结论：一定浓度的CGRP对大鼠成骨细胞转录因子RUNX2的表达有直接促进作用,RUNX2可能参与了CGRP刺激成骨细胞增殖反应的机制。

英文摘要：

Objective：To investigate the effects of exogenous calcitonin gene-related peptide（CGRP） on RUNX2 expression in primary cultured rat osteoblasts.Methods： Primary cultured rat osteoblasts were obtained by collagenase / trypsinenzyme digestion from calvarial bone of neonate rats and were identified by ALP staining;The cells were exposed to gradient concentrations,and the effect of CGRP on the proliferation of osteoblasts was determined by MTT assay.The expressions of RUNX2 were quantified by RT-PCR and Western blotting after 48-hour exposure to different CGRP concentrations.Results： CGRP could promote the proliferation of osteoblasts.When the concentration of CGRP were at 10-8-10-6 mol/L,the proliferation rates of osteoblasts were 71.9%,142.1%,321.0%,respectively（P 0.05）;when CGRP was at concentrations of 10-7 and 10-6 mol / L in osteoblasts for 48 h,the mRNA levels of RUNX2 was significantly increased,which increased（46.2 ± 11.2）% and（58.6 ± 14.0）% respectively（P 0.05）;the expression of RUNX2 protein levels was consistent with the change of the mRNA levels.Conclusion： CGRP could promote RUNX2 expression in primary cultured rat osteoblasts,which suggests that RUNX2 is involved in the mechanism of osteoblast proliferation induced by CGRP.