Department of Trauma Surgery, University of Duisburg-Essen, Hufelandstraße 55, 45147 Essen, Germany.
Int J Biol Sci. 2010 Sep 15;6(6):525-36. doi: 10.7150/ijbs.6.525.
A linkage between the neurotransmitter alpha-calcitonin gene-related peptide (alpha-CGRP) and particle-induced osteolysis has been shown previously. The suggested osteoprotective influence of alpha-CGRP on the catabolic effects of ultra-high molecular weight polyethylene (UHMWPE) particles is analyzed in this study in primary human osteoblasts.
Primary human osteoblasts were stimulated by UHMWPE particles (cell/particle ratios 1:100 and 1:500) and different doses of alpha-CGRP (10(-7 )M, 10(-9 )M, 10(-11 )M). Receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG) mRNA expression and protein levels were measured by RT-PCR and Western blot.
Particle stimulation leads to a significant dose-dependent increase of RANKL mRNA in both cell-particle ratios and a significant down-regulation of OPG mRNA in cell-particle concentrations of 1:500. A significant depression of alkaline phosphatase was found due to particle stimulation. Alpha-CGRP in all tested concentrations showed a significant depressive effect on the expression of RANKL mRNA in primary human osteoblasts under particle stimulation. Comparable reactions of RANKL protein levels due to particles and alpha-CGRP were found by Western blot analysis. In cell-particle ratios of 1:100 after 24 hours the osteoprotective influence of alpha-CGRP reversed the catabolic effects of particles on the RANKL expression.
The in-vivo use of alpha-CGRP, which leads to down-regulated RANKL in-vitro, might inhibit the catabolic effect of particles in conditions of particle induced osteolysis.
先前已经证明，神经递质α-降钙素基因相关肽（α-CGRP）与颗粒诱导性骨溶解之间存在关联。本研究旨在分析 α-CGRP 对超高分子量聚乙烯（UHMWPE）颗粒的分解代谢作用的保护作用，方法是在原代人成骨细胞中进行研究。
用人 UHMWPE 颗粒（细胞/颗粒比为 1:100 和 1:500）和不同剂量的 α-CGRP（10^(-7) M、10^(-9) M、10^(-11) M）刺激原代人成骨细胞。通过 RT-PCR 和 Western blot 测量核因子-κB 配体受体激活剂（RANKL）和骨保护素（OPG）的 mRNA 表达和蛋白水平。
颗粒刺激导致两种细胞/颗粒比的 RANKL mRNA 呈显著的剂量依赖性增加，而在细胞/颗粒浓度为 1:500 时，OPG mRNA 的表达显著下调。由于颗粒刺激，碱性磷酸酶显著降低。在颗粒刺激下，α-CGRP 在所有测试浓度下均显示出对原代人成骨细胞中 RANKL mRNA 表达的显著抑制作用。通过 Western blot 分析发现颗粒和 α-CGRP 引起的 RANKL 蛋白水平的相似反应。在 24 小时的细胞/颗粒比为 1:100 的情况下，α-CGRP 的骨保护作用逆转了颗粒对 RANKL 表达的分解代谢作用。
体内使用 α-CGRP 可导致体外 RANKL 下调，可能抑制颗粒诱导性骨溶解条件下颗粒的分解代谢作用。
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