卵巢切除小鼠骨髓间充质干细胞RANKL、OPG促进破骨细胞发育并增强其功能  

RANKL and OPG expression in bone marrow-derived mesenchymal stem cells of ovariectomied promotes osteoclast development and enhances its function

作　　者：邵秉一[1,2] 于洋[1,2] 傅潇慧[3] 薛红蕾[1,2] 戚朦[4] 帅逸[4] 周志斐[4] 金岩[4] 杨德琴[1,2] 

机构地区：[1]重庆医科大学附属口腔医院牙体牙髓科,重庆400015 [2]口腔疾病与生物医学重庆市重点实验室,重庆400015 [3]浙江大学医学院附属第二医院滨江院区,浙江杭州310009 [4]第四军医大学组织工程中心,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2013年第12期1262-1266,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家重点基础研究发展计划(973)(2011CB964700);重庆市医学重点学科建设经费基金资助项目《牙体牙髓病学》(2011年)

摘　　要：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 expression of receptor activator of nuclear factor-kappa B lig （RANKL）, osteo-protegerin （OPG） in bone marrow mesenchymal stem cells （BMSCs） and its impact on osteoclast formation and function inthe ovariectomied mice. Methods An animal model of osteoporosis was established by ovariectomy （OVX bilateral ovarianresection） in 8-week-old healthy female mice. The sham group was the 8-week-old healthy female mice with bilateralresection. Macrophages from mice were inducted by M-CSF and RANKL, and co-cultured with the BMSCs collected frommice in the OVX group and sham group, respectively. The osteoclast numbers of the two groups were compared by TRAPstaining. The resorption pits were measured by toluidine blue staining. The level of RANKL/OPG expression was detected byRT-PCR and Western blotting. Results TRAP assay and toluidine blue staining showed that the numbers of osteoclasts andresorption pits in OVX group were more than that in the sham group. The expression of RANKL in BMSC was lower in shamgroup than that of the OVX group. On the contrary, the expression of OPG in BMSCs was higher in the shame group thanthat of the OVX group. Conclusion Expression of RANKL/OPG are regulated by estrogen in BMSCs. RANKL and OPGexpression promotes osteoclast development and enhances its function under the condition of estrogen deficiency.

关 键 词：雌激素 骨质疏松 骨髓间充质干细胞 RANKL OPG 破骨细胞 

分 类 号：R392-33[医药卫生—免疫学] R681[医药卫生—基础医学]