Department of Internal Medicine, Endocrine Section, VU University Medical Center, Amsterdam, The Netherlands.
PLoS One. 2013 Nov 8;8(11):e79672. doi: 10.1371/journal.pone.0079672. eCollection 2013.
Skeletal integrity in humans and animals is maintained by daily mechanical loading. It has been widely accepted that osteocytes function as mechanosensors. Many biochemical signaling molecules are involved in the response of osteocytes to mechanical stimulation. The aim of this study was to identify genes involved in the translation of mechanical stimuli into bone formation. The four-point bending model was used to induce a single period of mechanical loading on the right tibia, while the contra lateral left tibia served as control. Six hours after loading, the effects of mechanical loading on gene-expression were determined with microarray analysis. Protein expression of differentially regulated genes was evaluated with immunohistochemistry. Nine genes were found to exhibit a significant differential gene expression in LOAD compared to control. MEPE, Garnl1, V2R2B, and QFG-TN1 olfactory receptor were up-regulated, and creatine kinase (muscle form), fibrinogen-B beta-polypeptide, monoamine oxidase A, troponin-C and kinesin light chain-C were down-regulated. Validation with real-time RT-PCR analysis confirmed the up-regulation of MEPE and the down-regulation of creatine kinase (muscle form) and troponin-C in the loaded tibia. Immunohistochemistry showed that the increase of MEPE protein expression was already detectable six hours after mechanical loading. In conclusion, these genes probably play a role during translation of mechanical stimuli six hours after mechanical loading. The modulation of MEPE expression may indicate a connection between bone mineralization and bone formation after mechanical stimulation.
在人类和动物中,骨骼的完整性是通过日常的机械负荷来维持的。骨细胞作为机械感受器的功能已被广泛接受。许多生化信号分子参与了骨细胞对机械刺激的反应。本研究旨在鉴定将机械刺激转化为骨形成的相关基因。四点弯曲模型用于对右侧胫骨施加单次机械加载,而对侧左侧胫骨作为对照。在加载 6 小时后,通过微阵列分析确定机械加载对基因表达的影响。采用免疫组织化学法评估差异调节基因的蛋白表达。与对照组相比,在 LOAD 中发现 9 个基因的表达存在显著差异。MEPE、Garnl1、V2R2B 和 QFG-TN1 嗅觉受体上调,而肌型肌酸激酶、纤维蛋白原-B 亚基多肽、单胺氧化酶 A、肌钙蛋白-C 和驱动蛋白轻链-C 下调。实时 RT-PCR 分析的验证证实了 MEPE 的上调和负载胫骨中肌型肌酸激酶和肌钙蛋白-C 的下调。免疫组织化学显示,MEPE 蛋白表达的增加在机械加载后 6 小时即可检测到。总之,这些基因可能在机械加载后 6 小时机械刺激的翻译过程中发挥作用。MEPE 表达的调节可能表明机械刺激后骨矿化和骨形成之间存在联系。