Strontium ranelate inhibits key factors affecting bone remodeling in human osteoarthritic subchondral bone osteoblasts
Strontium ranelate inhibits key factors affecting bone remodeling in human osteoarthritic subchondral bone osteoblasts
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DOI：
10.1016/j.bone.2011.06.005
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发表时间：
2011-09-01
期刊：
影响因子：
4.1
通讯作者：
Martel-Pelletier, Johanne
中科院分区：
文献类型：
--
作者：
Tat, Steeve Kwan;Pelletier, Jean-Pierre;Martel-Pelletier, Johanne
在骨关节炎（OA）中，软骨退变的进展与软骨下骨的重塑有关。人类OA软骨下骨成骨细胞表现出异常表型和代谢改变，导致异常的吸收过程。骨吸收的发生，至少部分是通过两种蛋白水解酶，MMP-2和MMP-9，以及RANKL水平的增加，这两种酶主要由成骨细胞产生。在本研究中，我们研究了雷乃酸锶对人OA软骨下成骨细胞上述关键因素的调节作用。方法：人软骨下成骨细胞在含0.1、1、2 mM雷奈酸锶的培养基中培养18 h，测定mRNA和蛋白含量72 h。评价雷奈酸钠对MMP-2、MMP-9、OPG、RANKL（总）、RANKL-1、RANKL-3表达的影响（VCR），对OPG （ELISA）、膜状RANKL（流式细胞术）、MT1-MMP、ADAM17、ADAM19 （Western blot）产生的影响。成骨细胞与前破骨细胞（即分化的人外周血单个核细胞）孵育后，用亚微米合成磷酸钙薄膜测量吸收表面。结果：首先检测正常和OA软骨下成骨细胞中MMP-2、MMP-9、OPG、RANKL的表达水平。正如预期的那样，正常细胞中检测不到MMP-9和RANKL的基因表达，而MMP-2的表达非常低，但可以检测到，并且OPG表现出高基因表达。进一步的实验观察雷奈酸锶对表达水平的影响，除了OPG，只在OA软骨下骨成骨细胞上进行。在OA细胞中，1mm雷奈酸锶显著降低了MMP-2和MMP-9的表达水平(p
Introduction: In osteoarthritis (OA) the progression of cartilage degeneration has been associated with remodeling of the subchondral bone. Human OA subchondral bone osteoblasts were shown to have an abnormal phenotype and altered metabolism leading to an abnormal resorptive process. Bone resorption is suggested to occur, at least in part, through the increased levels of two proteolytic enzymes, MMP-2 and MMP-9, and RANKL, which are mainly produced by osteoblasts. In this study, we investigated in human OA subchondral bone osteoblasts the modulatory effect of strontium ranelate on the above key factors.Methods: Human subchondral bone osteoblasts were cultured in a medium containing 0.1, 1 and 2 mM of strontium ranelate for 18 h for mRNA and 72 h for protein determination. The effect of strontium ranelate was evaluated on the expression ((VCR) of MMP-2, MMP-9, OPG, RANKL (total), RANKL-1, and RANKL-3, on the production of OPG (ELISA), membranous RANKL (flow cytometry), and MT1-MMP, ADAM17, and ADAM19 (Western blot). After incubation of osteoblasts with pre-osteoclasts (i.e., differentiated human peripheral blood mononuclear cells), the resorbed surface was measured using a sub-micron synthetic calcium phosphate thin film.Results: Firstly, the expression levels of MMP-2, MMP-9, OPG, and RANKL were determined in normal and OA subchondral bone osteoblasts. As expected, the gene expression of MMP-9 and RANKL were not detectable in normal cells, whereas MMP-2 was very low but detectable and OPG demonstrated high gene expression. Further experiments looking at the effect of strontium ranelate on expression levels, except for OPG, were performed only on the OA subchondral bone osteoblasts. In OA cells, the expression levels of MMP-2 and MMP-9 were significantly decreased by strontium ranelate at 1 mM (p