Takano-Yamamoto T, Rodan G A
Department of Bone Biology and Osteoporosis Research, Merck Sharp & Dohme Research Laboratories, West Point, PA 19486.
Proc Natl Acad Sci U S A. 1990 Mar;87(6):2172-6. doi: 10.1073/pnas.87.6.2172.
High-affinity nuclear binding sites for 17 beta-estradiol (17 beta E2) were recently found in bone cells; however, the mechanism by which estrogen exerts its effect on bone in vivo is still unknown. To study if estrogen acts on bone directly, we used an experimental model in which test substances are infused locally into rat femur trabecular bone. Sprague-Dawley rats weighing 150-160 g were ovariectomized (OVX) and 14 days later a polyethylene tube (1 mm in diameter) connected to an Alzet osmotic minipump was implanted into the distal femur 9 mm from the joint. 17 beta E2 (24 microliters/day at 0.01-1 nM), 17 alpha-estradiol (17 alpha E2) (24 microliters/day at 1 nM), or phosphate-buffered saline (NaCl, 8 g/liter; KCl, 0.2 g/liter; KH2PO4, 0.2 g/liter; Na2HPO4.7H2O, 2.16 g/liter) was infused for 8 days. The contralateral limb remained intact. Animals were sacrificed and bones were examined by histomorphometry. Ovariectomy caused a 50% loss in trabecular bone volume (TBV) in the secondary spongiosa (from 20.3% +/- 1.7% to 9.6% +/- 1.1%; mean +/- SEM), a 2-fold increase in osteoclast number (to 4.0 +/- 0.4 per mm), a 3-fold increase in relative resorption surfaces (to 24.8% +/- 2.9%), a 9-fold increase in osteoblast number (to 11.3 +/- 2.1 per mm), and an 8-fold increase in relative osteoid surface (to 9.6% +/- 1.7%). The local infusion of 17 beta E2 for 8 days into OVX rats (i) restored the TBV dose dependently to 75% and 85% of control (non-OVX) levels, at 0.1 nM and 1 nM 17 beta E2, respectively; (ii) decreased osteoclast number and the relative resorption surface to control (non-OVX) levels; and (iii) further increased osteoblast number and the relative osteoid surface dose dependently (by 5-fold at 1 nM 17 beta E2). Phosphate-buffered saline infusion was without effect. Infusion of 17 alpha E2 had no effect on TBV, osteoclast number, or resorption surface but increased slightly the osteoblast number and the osteoid surface. Its potency was 1/100 that of 17 beta E2. The local infusion of 17 beta E2 or 17 alpha E2 had no effect on body or uterine weight. We conclude from these findings that estrogen delivered directly to the bone of OVX rats in vivo at 2.4 and 24 fmol/day acted locally to inhibit bone resorption and stimulate bone formation.
最近在骨细胞中发现了17β-雌二醇(17β-E2)的高亲和力核结合位点;然而,雌激素在体内对骨骼发挥作用的机制仍不清楚。为了研究雌激素是否直接作用于骨骼,我们使用了一种实验模型,即将受试物质局部注入大鼠股骨小梁骨。将体重150 - 160克的Sprague-Dawley大鼠进行卵巢切除术(OVX),14天后,将连接到Alzet渗透微型泵的聚乙烯管(直径1毫米)植入距关节9毫米的股骨远端。注入17β-E2(0.01 - 1 nM,每天24微升)、17α-雌二醇(17α-E2)(1 nM,每天24微升)或磷酸盐缓冲盐水(NaCl,8克/升;KCl,0.2克/升;KH2PO4,0.2克/升;Na2HPO4·7H2O,2.16克/升),持续8天。对侧肢体保持完整。处死动物后,通过组织形态计量学检查骨骼。卵巢切除术导致次级海绵体中的小梁骨体积(TBV)减少50%(从20.3%±1.7%降至9.6%±1.1%;平均值±标准误),破骨细胞数量增加2倍(至每毫米4.0±0.4个),相对吸收表面增加3倍(至24.8%±2.9%),成骨细胞数量增加9倍(至每毫米11.3±2.1个),相对类骨质表面增加8倍(至9.6%±1.7%)。对OVX大鼠局部注入17β-E2 8天,(i)在0.1 nM和1 nM 17β-E2时,分别将TBV剂量依赖性地恢复至对照(非OVX)水平的75%和85%;(ii)将破骨细胞数量和相对吸收表面降至对照(非OVX)水平;(iii)剂量依赖性地进一步增加成骨细胞数量和相对类骨质表面(在1 nM 17β-E2时增加5倍)。注入磷酸盐缓冲盐水没有效果。注入17α-E2对TBV、破骨细胞数量或吸收表面没有影响,但略微增加了成骨细胞数量和类骨质表面。其效力为17β-E2的1/100。局部注入17β-E2或17α-E2对体重和子宫重量没有影响。我们从这些发现中得出结论,以每天2.4和24飞摩尔的剂量直接输送到OVX大鼠体内骨骼的雌激素在局部发挥作用,抑制骨吸收并刺激骨形成。