破骨细胞骨吸收上清液对骨髓间充质干细胞增殖分化的影响  

Effect of the supernatant of osteoclasts during bone resorption on the proliferation and differentiation of bone marrow-derived mesenchymal stem cells

作　　者：邵秉一[1,2] 于洋[1,2] 蒋程程[3] 帅逸[4] 戚朦[4] 景欢[4] 尚冯青[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 [2]口腔疾病与生物医学重庆市重点实验室,重庆400015 [3]杭州市红十字会医院口腔科,浙江杭州310003 [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》2014年第3期241-244,249,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<牙体牙髓病学>(2011年)

摘　　要：目的研究破骨细胞骨吸收活动中的分泌产物对骨髓间充质干细胞增殖、分化的影响。方法诱导小鼠脾脏细胞为破骨细胞,用抗酒石酸盐酸性磷酸酶(TRAP)染色。破骨细胞与牛骨磨片共培养,扫描电镜观察骨吸收陷窝。收集骨吸收实验破骨细胞培养上清液作用于小鼠骨髓间充质干细胞(BMSC),MTT法检测BMSC生长曲线;成骨诱导后钙化结节茜素红染色法检测BMSC成骨能力;成脂诱导后油红O染色检测BMSC成脂能力;Western blot法检测小鼠BMSC成骨相关蛋白RUNX2、碱性磷酸酶(ALP)及成脂相关蛋白过氧化物酶体增殖物激活受体γ(PPAR-γ)的表达。结果 TRAP染色、扫描电镜显示脾脏细胞可诱导分化为具有骨吸收能力的破骨细胞;与对照组相比,加入破骨细胞培养上清液,BMSC的增殖受到抑制,成骨分化增强,成脂分化减弱(P<0.05)。结论破骨细胞骨吸收上清液具有使BMSC增殖能力降低,成骨分化增强,成脂分化减弱的作用。Objective To investigate the effect of the supernatant of osteoclasts during bone resorption on the proliferation and differentiation of bone marrow-derived mesenchymal stem cells （BMSCs）. Methods Spleen cells were induced into osteoclasts which were identified with tartrate-resistant acid phosphatase （TRAP） staining. The osteoclasts were cultured with bovine bone slices and the resorption pits on the slices were observed by scanning electron microscopy. The supernatant of the osteoclasts during bone absorption were taken and added into BMSCs. M-I-F assay was used to analyze the growth curves of BMSCs in experimental and control groups. After adipogenic induction, lipid droplets were observed by oil red O staining to compare the adipogenic potential between the two groups. After osteogenic induction, calcium nodules were observed by alizarin red staining. The expression levels of osteogenesis-related proteins RUNX2 and alkaline phosphatase （ALP）, and adipogenesis-related protein PPAR-y were measured by Western blotting. Results TRAP staining and scanning electron microscopy indicated that BMSCs could be differentiated into functional osteoclasts. Compared with the control group, the proliferation capacity of BMSCs in the experimental group was suppressed, the osteogenic potential was enhanced, and the adipogenic potential was weakened. Conclusion The supernatant of osteoclasts during bone absorption might increase the osteogenic activity of BMSCs by inhibiting proliferation and adipogenesis and promoting differentiation.

关 键 词：破骨细胞 骨髓来源的间充质干细胞 增殖 成骨成脂 

分 类 号：R392.12[医药卫生—免疫学] Q254[医药卫生—基础医学]