Xu Jie, Kauther Max D, Hartl Julia, Wedemeyer Christian
Department of Orthopaedics, University of Duisburg-Essen, Pattbergstrasse 1-3, 45239 Essen, Germany.
J Orthop Surg Res. 2010 Nov 4;5:83. doi: 10.1186/1749-799X-5-83.
Recent studies demonstrated an impact of the nervous system on particle-induced osteolysis, the major cause of aseptic loosening of joint replacements.
In this study of MG-63 osteoblast-like cells we analyzed the influence of ultra-high molecular weight polyethylene (UHMWPE) particles and the neurotransmitter alpha-calcitonin gene-related peptide (CGRP) on the osteoprotegerin/receptor activator of nuclear factor-κB ligand/receptor activator of nuclear factorκB (OPG/RANKL/RANK) system. MG-63 cells were stimulated by different UHMWPE particle concentrations (1:100, 1:500) and different doses of alpha-CGRP (10-7 M, 10-9 M, 10-11 M). RANKL and OPG mRNA expression and protein levels were measured by RT-PCR and Western blot.
Increasing particle concentrations caused an up-regulation of RANKL after 72 hours. Alpha-CGRP showed a dose-independent depressive effect on particle-induced expression of RANKL mRNA in both cell-particle ratios. RANKL gene transcripts were significantly (P < 0.05) decreased by alpha-CGRP treatment after 48 and 72 hours. OPG mRNA was significantly down-regulated in a cell-particle ratio of 1:500 after 72 hours. Alpha-CGRP concentrations of 10-7 M lead to an up-regulation of OPG protein.
In conclusion, a possible osteoprotective influence of the neurotransmitter alpha-CGRP on particle stimulated osteoblast-like cells could be shown. Alpha-CGRP might be important for bone metabolism under conditions of particle-induced osteolysis.
近期研究表明,神经系统对颗粒诱导的骨溶解有影响,而颗粒诱导的骨溶解是关节置换无菌性松动的主要原因。
在这项针对MG-63成骨细胞样细胞的研究中,我们分析了超高分子量聚乙烯(UHMWPE)颗粒和神经递质α-降钙素基因相关肽(CGRP)对骨保护素/核因子κB受体活化因子配体/核因子κB受体活化因子(OPG/RANKL/RANK)系统的影响。MG-63细胞分别用不同浓度的UHMWPE颗粒(1:100、1:500)和不同剂量的α-CGRP(10-7 M、10-9 M、10-11 M)进行刺激。通过逆转录聚合酶链反应(RT-PCR)和蛋白质印迹法检测RANKL和OPG的mRNA表达及蛋白水平。
颗粒浓度增加导致72小时后RANKL上调。α-CGRP在两种细胞与颗粒比例下均对颗粒诱导的RANKL mRNA表达呈现剂量非依赖性抑制作用。α-CGRP处理48小时和72小时后,RANKL基因转录物显著减少(P < 0.05)。72小时后,在细胞与颗粒比例为1:500时,OPG mRNA显著下调。10-7 M的α-CGRP浓度导致OPG蛋白上调。
总之,可证明神经递质α-CGRP对颗粒刺激的成骨细胞样细胞可能具有骨保护作用。在颗粒诱导的骨溶解情况下,α-CGRP可能对骨代谢很重要。