神经生长因子调控降钙素基因相关肽的表达及对MG-63细胞增殖的影响

Experimental study on the effects of the nerve growth factor regulating calcitonin gene-related peptide in promoting the proliferation of MG-63 in vitro

目的探讨创伤愈合过程中神经生长因子(NGF)调控降钙素基因相关肽(CGRP)的表达,以及影响MG-63细胞增殖的作用机制。方法加入不同质量浓度的NGF刺激MG-63细胞,1、2、3、4 d后收集样本,采用酶联免疫吸附测定法(ELISA)检测CGRP的表达,实时荧光定量聚合酶链反应(RT-QPCR)检测CGRP m RNA的表达,采用细胞计数法(CCK-8)检测MG-63细胞的增殖;在MG-63细胞中加入NGF受体阻断剂,采用RT-QPCR和CCK-8检测CGRP m RNA的表达及MG-63细胞的增殖效率。结果在NGF作用下,MG-63细胞分泌的CGRP表达量明显上调,随NGF质量浓度升高,CGRP表达量也相应升高,具有浓度依赖性,CGRP表达量随NGF作用时间延长也相应增加(P<0.05);随NGF质量浓度升高和作用时间延长,MG-63细胞的增殖效率也相应增加(P<0.05)。加入NGF受体阻断剂后此作用相应减弱(P<0.05)。结论在创伤愈合过程中NGF能通过上调CGRP的表达量影响MG-63细胞的增殖,从而促进创伤愈合。

ObjectiveTo investigate the nerve growth factor （NGF） regulating the expression of calcitonin gene-related peptide （CGRP） in promoting the proliferation of osteoblast-like cell （MG-63） and thus illustrate the mechanism of the NGF in wound healing. Methods Different concentrations of NGF were used to stimulate MG-63. The expression of CGRP was detected by real-time quantitative polymerase chain reaction （RT-QPCR） and enzyme-linked immunosorbent assay after 1, 2, 3, and 4 days. The proliferation of MG-63 was detected by cell counting kit-8 （CCK-8）. The expression of CGRP mRNA and the proliferation of MG-63 were then detected by RT-QPCR and CCK-8 after adding the NGF receptor blocker. Results Com-pared with the blank control group, the expression of CGRP significantly increased by stimulating the NGF. The expression of CGRP was positively related to the concentration of NGF （P〈0.05）. Moreover, the expression of CGRP increased by pro-longing the NGF stimulation time. The proliferation of MG-63 increased after stimulating the NGF （P〈0.05）. After adding the NGF receptor blocker, the expression of CGRP and the proliferation of MG-63 correspondingly decreased （P〈0.05）. Conclusion NGF can up-regulate the expression of CGRP and increase the proliferation of MG-63. Therefore, NGF plays a significant role in wound healing.