In vitro expression of receptor activator of nuclear factor-kappaB ligand and osteoprotegerin in cultured equine articular cells.
In vitro expression of receptor activator of nuclear factor-kappaB ligand and osteoprotegerin in cultured equine articular cells.
复制标题
核因子-κB 配体受体激活剂和骨保护素在培养的马关节细胞中的体外表达。
DOI：
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发表时间：
2010
影响因子：
1
通讯作者：
T. Fan
中科院分区：
文献类型：
--
作者：
C. Byron;A. Barger;A. Stewart;H. Pondenis;T. Fan
目的
测定马软骨细胞和滑膜细胞中核因子-κ B配体受体激活剂（RANKL）和骨保护素（OPG）的浓度，并定量OPG：RANKL比值响应外源性因素的变化。
样本群体
从5匹成年（1- 8岁）马的掌指关节和跖趾关节获得外观大体正常的关节软骨和滑膜样本。
程序
将软骨细胞和滑膜细胞的细胞培养物与人重组白细胞介素-1 β（hrIL-1 β; 10 ng/mL）、脂多糖（LPS; 10 μ g/mL）或地塞米松（100 nM）孵育48小时。阴性对照培养物未接受处理。通过使用蛋白质印迹和免疫细胞化学分析测定细胞和用过的培养基的RANKL和OPG浓度。还通过使用ELISA测定用过的培养基的OPG浓度。
结果
RANKL和OPG在体外培养的马软骨细胞和滑膜细胞中均有表达。细胞相关RANKL和OPG浓度不受外源性因素的影响。软骨细胞中hrIL-1 β显著增加了培养基中可溶性RANKL的释放，但在滑膜细胞培养物中没有增加。hrIL-1 β和LPS可显著增加软骨细胞中可溶性OPG向培养基中的释放，但在滑膜细胞培养物中无此作用。软骨细胞培养物中的可溶性OPG：RANKL比率被LPS显著增加。地塞米松降低滑膜细胞OPG表达。
结论和临床相关性
RANKL和OPG蛋白在马关节细胞中表达。这些蛋白质的释放可能会影响邻近软骨下骨内的破骨细胞生成。因此，RANKL和OPG可用作关节疾病马的生物标志物和治疗靶点。
OBJECTIVE
To determine concentrations of receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) in equine chondrocytes and synoviocytes and to quantify changes in the OPG:RANKL ratio in response to exogenous factors.
SAMPLE POPULATION
Samples of articular cartilage and synovium with grossly normal appearance obtained from metacarpophalangeal and metatarsophalangeal joints of 5 adult (1- to 8-year-old) horses.
PROCEDURES
Cell cultures of chondrocytes and synoviocytes were incubated with human recombinant interleukin-1beta (hrIL-1beta; 10 ng/mL), lipopolysaccharide (LPS; 10 microg/mL), or dexamethasone (100nM) for 48 hours. Negative control cultures received no treatment. Cells and spent media were assayed for RANKL and OPG concentrations by use of western blot and immunocytochemical analyses. Spent media were also assayed for OPG concentration by use of an ELISA.
RESULTS
RANKL and OPG were expressed in equine chondrocytes and synoviocytes in vitro. Cell-associated RANKL and OPG concentrations were not impacted by exogenous factors. Soluble RANKL release into media was significantly increased by hrIL-1beta in chondrocyte but not in synoviocyte cultures. Soluble OPG release into media was significantly increased by hrIL-1beta and LPS in chondrocyte but not in synoviocyte cultures. The soluble OPG:RANKL ratio was significantly increased by LPS in chondrocyte cultures. Dexamethasone decreased OPG expression in synoviocytes.
CONCLUSIONS AND CLINICAL RELEVANCE
RANKL and OPG proteins were expressed in equine articular cells. Release of these proteins may affect osteoclastogenesis within adjacent subchondral bone. Thus, RANKL and OPG may have use as biomarkers and treatment targets in horses with joint disease.