Synthesis of osteoprotegerin and RANKL by megakaryocytes is modulated by oestrogen
Synthesis of osteoprotegerin and RANKL by megakaryocytes is modulated by oestrogen
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DOI：
10.1111/j.1365-2141.2004.05024.x
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发表时间：
2004-07-01
影响因子：
6.5
通讯作者：
Compston, JE
中科院分区：
文献类型：
--
作者：
Bord, S;Frith, E;Compston, JE
为了研究巨核细胞（MK）影响骨重塑的机制，将CD34（+）细胞在有或没有17β-雌二醇（E）的情况下培养6、9和12天，并对骨保护素（OPG）、核因子（NF）-κB配体受体激活剂（RANKL）和CD61进行免疫定位。通过图像分析定量测量特定蛋白质表达。基于荧光的免疫细胞化学用于将 OPG 和 RANKL 与 CD61 共定位。在 24 和 48 小时时，在有或没有 E 的情况下评估 CD61(+) 细胞中的 OPG 和 RANKL mRNA。 6 天时，E 处理后 OPG 和 RANKL 表达没有变化。第 9 天，经过 E 处理的具有成熟 MK 的培养物显示 OPG 表达增加 1.72 倍（P < 0.01），RANKL 减少 1.8 倍（P < 0.01）。 12 天时 OPG 表达达到最大，表达诱导增加了三倍（P < 0.001），而 RANKL 水平与对照相比进一步抑制了 2.3 倍（P < 0.001）。 CD61 与 OPG 和 RANKL 共定位。 mRNA 数据与蛋白质数据一致，E 将 OPG 表达诱导 90 倍，将 RANKL 表达抑制 34 倍（P < 0.001）。因此，E 刺激巨核细胞生成并调节 OPG 和 RANKL 表达，这提供了 MK 可能在骨重塑中发挥作用的证据，特别是在 E 诱导的破骨细胞生成和骨吸收变化中。
To investigate the mechanisms by which megakaryocytes (MKs) may influence bone remodelling, CD34(+) cells were cultured for 6, 9 and 12 d with or without 17beta-oestradiol (E) and immunolocalized for osteoprotegerin (OPG), receptor activator of nuclear factor (NF)-kappaB ligand (RANKL) and CD61. Specific protein expression was measured quantitatively by image analysis. Fluorescence-based immunocytochemistry was used to co-localize OPG and RANKL with CD61. OPG and RANKL mRNA was assessed in CD61(+) cells with or without E at 24 and 48 h. At 6 d, OPG and RANKL expression was unchanged by E treatment. At 9 d, the E-treated cultures with maturing MKs showed a 1.72-fold (P < 0.01) increase in OPG expression and a 1.8-fold (P < 0.01) reduction in RANKL. Maximal OPG expression was seen at 12 d with a threefold induction of expression (P < 0.001), whilst RANKL levels were further suppressed by 2.3-fold compared with controls (P < 0.001). CD61 co-localized with OPG and RANKL. mRNA data were consistent with that of protein, with a 90-fold induction in OPG expression and a 34-fold suppression of RANKL expression by E (P < 0.001). Thus, E stimulates megakaryocytopoiesis and modulates OPG and RANKL expression, providing evidence that MKs may play a role in bone remodelling and, in particular, in E-induced changes in osteoclastogenesis and bone resorption.