de Souza Malaspina Tatiana Salles, dos Santos Célio Xavier, Campanelli Ana Paula, Laurindo Francisco Rafael Martins, Sogayar Mari Cleide, Granjeiro José Mauro
Department of Biological Sciences, Bauru Dental School-University of São Paulo, Bauru, SP, Brazil.
J Mol Histol. 2008 Dec;39(6):627-34. doi: 10.1007/s10735-008-9203-0. Epub 2008 Nov 1.
Tartrate-resistant acid phosphatase (TRAP) is a well-known marker of osteoclasts and bone resorption. Here we have investigated whether osteoblast-like cells (hFOB 1.19) present TRAP activity and how would be its pattern of expression during osteoblastic differentiation. We also observed how the osteoblastic differentiation affected the reduced glutathione levels. TRAP activity was measured using the p-nitrophenylphosphate substrate. The osteogenic potential of hFOB 1.19 cells was studied by measuring alkaline phosphatase activity and mineralized nodule formation. Oxidative stress was determined by HPLC and DNTB assays. TRAP activity and the reduced glutathione-dependent microenvironment were modulated during osteoblastic differentiation. During this phase, TRAP activity, as well as alkaline phosphatase and glutathione increased progressively up to the 21st day, decreasing thereafter. We demonstrate that TRAP activity is modulated during osteoblastic differentiation, possibly in response to the redox state of the cell, since it seemed to depend on suitable levels of reduced glutathione.
抗酒石酸酸性磷酸酶（TRAP）是破骨细胞和骨吸收的著名标志物。在此，我们研究了成骨样细胞（hFOB 1.19）是否具有TRAP活性，以及在成骨细胞分化过程中其表达模式如何。我们还观察了成骨细胞分化如何影响还原型谷胱甘肽水平。使用对硝基苯磷酸底物测量TRAP活性。通过测量碱性磷酸酶活性和矿化结节形成来研究hFOB 1.19细胞的成骨潜能。通过高效液相色谱法（HPLC）和二硝基苯磺酸（DNTB）测定法确定氧化应激。在成骨细胞分化过程中，TRAP活性和依赖于还原型谷胱甘肽的微环境受到调节。在此阶段，TRAP活性以及碱性磷酸酶和谷胱甘肽在第21天之前逐渐增加，此后下降。我们证明，TRAP活性在成骨细胞分化过程中受到调节，可能是对细胞氧化还原状态的反应，因为它似乎依赖于适当水平的还原型谷胱甘肽。
