论文部分内容阅读
目的研究雌激素调控骨髓间充质干细胞(BMSC)表达核因子κB受体活化因子配体(RANKL)、骨保护因子(OPG)对破骨细胞发育和功能的影响。方法选用健康雌性小鼠行双侧卵巢切除术(OVX),建立绝经后骨质疏松模型。选用同一批次健康小鼠行双侧卵巢脂肪组织部分切除,建立假手术组(sham)。用巨噬细胞集落刺激因子(M-CSF)、RANKL诱导小鼠单核细胞为破骨细胞,同时分别加入sham和OVX组BMSC与单核细胞共培养。抗酒石酸酸性磷酸酶(TRAP)染色计数2组BMSC对骨髓诱导发育为破骨细胞的数量。甲苯胺蓝染色检测2组共培养体系中破骨细胞骨吸收陷窝的数量。RT-PCR和Western blot法检测sham和OVX组BMSC中OPG、RANKL的表达。结果 TRAP和甲苯胺蓝染色显示,OVX组中破骨细胞数目及吸收陷窝数大于sham组(P<0.05)。RT-PCR和Western blot法结果显示OVX组较sham组OPG表达显著降低,RANKL显著增强(P<0.05)。结论雌激素能够影响BMSC中OPG和RANKL的表达,雌激素缺乏环境下,BMSC促进破骨细胞发育并增强其功能。
Objective To investigate the effects of estrogen-regulated bone marrow mesenchymal stem cells (BMSCs) on the development and function of osteoclasts by expressing RANKL and osteoprotegerin (OPG). Methods Healthy female mice underwent bilateral ovariectomy (OVX) to establish a model of postmenopausal osteoporosis. Select the same batch of healthy mice bilateral ovarian adipose tissue resection, the establishment of sham group (sham). Macrophage colony stimulating factor (M-CSF) was used to induce the mononuclear cells of mice to be osteoclasts. At the same time, BMSCs were co-cultured with monocytes in sham and OVX groups respectively. Tartrate resistance acid phosphatase (TRAP) staining The number of bone marrow-induced osteoclasts was counted in two groups of BMSCs. Toluidine blue staining was used to detect the number of osteoclast resorption lacunae in two co-culture systems. The expression of OPG and RANKL in BMSC were detected by RT-PCR and Western blot in sham and OVX groups. Results TRAP and toluidine blue staining showed that the number of osteoclasts and the number of lacunae in OVX group were greater than those in sham group (P <0.05). The results of RT-PCR and Western blot showed that OPG expression in OVX group was significantly lower than sham group, and RANKL was significantly increased (P <0.05). Conclusion Estrogen can affect the expression of OPG and RANKL in BMSCs. In the absence of estrogen, BMSC promotes osteoclast development and enhances its function.