去卵巢大鼠骨髓源性破骨样细胞增殖、分化的改变及雌激素的影响

Changes of osteoclast-like cell proliferation and differentiation in bone marrow stem cell culture of ovariectomized rats and the effects of estrogen

目的 通过体外骨髓细胞培养诱导破骨样细胞 (osteoclast likecells ,OLC)的形成 ,观察去卵巢对成年大鼠OLC形成及活性的影响以及给予雌激素后的改变。 方法 3月龄SD大鼠分为对照组、去卵巢组及雌激素替代组。术后 12周处死大鼠 ,取股骨分离骨髓细胞 ,在条件培养液中诱导其向破骨细胞分化。活体观察OLC形成情况并于培养的第 6天行细胞染色 ,以抗酒石酸酸性磷酸酶(TRAP)染色 (+)、细胞核≥ 3个的细胞为OLC ,计数各组OLC及骨陷窝。 结果 3组中去卵巢组OLC出现早且数量〔(2 7 75± 0 92 )个 /玻片〕明显高于其它两组〔(17 93± 0 6 9)个 /玻片和 (12 81±0 6 1)个 /玻片 ,P <0 0 1〕。雌激素处理能明显抑制OLC的形成 ,但雌激素替代组的OLC仍多于对照组 (P <0 0 5 )。骨陷窝形成的变化与OLC数量改变一致。 结论 本实验结果显示大鼠去卵巢后 ,骨髓干细胞分化形成OLC数量明显增多 ,且活性增强。补充雌激素能够有效抑制OLC形成增加及其活性增强。故雌激素抑制骨吸收的机制至少部分是作用于骨髓干细胞向破骨细胞的分化。

Objective To investigate the effect of ovariectomy and estrogen replacement on the derivation of bone marrow stem cells to osteoclast-like cells (OLC) and the cell activity in rats by bone marrow cells culture in vitro. Methods 3-month-old SD rats were divided randomly into 3 groups: normal control (N), ovariectomized (OVX) and OVX plus estrogen treatment (OVX+E2). At the end of 12 week after ovariectomy, femur bone marrow were collected and cultured. At 6-th culture day, OLC was identified by the characteristics of TRAP staining (+) and three nuclei or more. The number of OLCs and bone resorption pits were measured under microscopy. Results Compare with N and OVX+E2, the number of OLCs and bone resorption pits in OVX was more(P<0.01)and cells appeared relatively earlier. This change was greatly inhibited by estrogen treatment(P<0.01), althought the number of OLCs and bone resorption pits in OVX+E2 was still more than in normal group(P<0.05). Conclusions The loss of ovary function increases the derivation of bone marrow cells to OLC and the cells activity, and estrogen replacement therapy can inhibit the effects. Estrogen plays an important role in the process of osteoclast-like multinucleated cell proliferation and differentiation from its progenitors at the cellular level.