雷奈酸锶对大鼠骨髓间充质干细胞增生和成脂分化的影响  

The effect of strontium ranelate on cell proliferation and adipogenic differentiation of rat mesenchymal stem cells

作　　者：王雪鹏[1] 郝永强[1] 曹磊[1] 陆耀刚[1] 金芳纯[1] 王磊[1] 

机构地区：[1]上海市骨科内植物重点实验室,上海交通大学医学院附属第九人民医院骨科,上海200011

出　　处：《中华骨质疏松和骨矿盐疾病杂志》2011年第1期36-43,共8页Chinese Journal Of Osteoporosis And Bone Mineral Research

基　　金：国家自然科学基金项目(81071472);国家高技术研究发展计划(863计划)(2009AA03Z311);上海市教育委员会重点学科建设基金(J50206);上海市骨科内植物重点实验室建设基金(08DZ2230330)

摘　　要：目的从细胞和基因水平研究不同浓度雷奈酸锶对大鼠骨髓间充质干细胞成脂分化的影响。方法分离培养并鉴定大鼠骨髓间充质干细胞。以不同浓度雷奈酸锶(0、0.05、0.1、0.5、1.0和5.0 mmol/L)刺激骨髓间充质干细胞,测定其增生活性。在成脂诱导条件下,加入不同浓度雷奈酸锶(0、1.0、5.0 mmol/L),倒置显微镜下检测油红染色并进行成脂相关基因检测。结果在非诱导条件下,浓度为1.0和5.0 mmol/L的雷奈酸锶对细胞增生具有明显促进作用。在成脂诱导6及9 d后,1.0和5.0 mmol/L雷奈酸锶均对细胞成脂分化具有抑制作用,表现为细胞内脂滴缩小,油红染色阳性细胞数量减低,成脂分化关键基因PPAR-γ、C/EBP-β、aP-2以及Glut-4的表达量降低。结论在体外雷奈酸锶具有促进骨髓间充质干细胞增生作用,并抑制其向脂肪细胞分化,这可能与其具有促进前体干细胞成骨分化并维持成骨与成脂分化平衡作用有关。Objective To study the effect of strontium ranelate at a series of concentrations on proliferation and adipogenesis of rat bone marrow mesenchymal stem cells from the cellular and genetic levels respectively. Methods Rat bone marrow mesenchymal stem cells were isolated, cultured and identified. MSCs were treated with in six experimental concentrations Strontium Ranelate （0 mmol/L, 0. 05 mmol/L, 0. 1 mmol/L, 0. 5 mmol/L, 1.0 mmol/L, 5.0mmol/L）, and cell proliferation was analyzed by MTF assay. Adipogenic differentiation was characterized by inverted phase contrast microscope, oil red O staining, inhibiting rate of adipogenesis and real-time PCR for adipocytes markers （PPAR-γ, C/EBP-β, aP-2 and Glut-4 ） and relative gene expression. Results In non-inducing conditions, strontium ranelate （ 1.0 mmol/L and 5.0 mmol/L） has significantly promoted MSCs proliferation. Adipogenic differentiation was inhibited by strontium ranelate （ 1.0 mmol/L and 5.0 mmol/L）. After 6 days or 9 days treated with strontium ranelate in adipo- genic medium, we found a significant deduction of Red O positive cells and intracellular lipid droplets compared to MSCs untreated strontium ranelate. In addition, the mRNA expression level of adipogenic differentiation key gene such as PPAR-γ / EBP-β, aP-2 and Glut-4 was reduced. Conclusion Strontium ranelate increased MSCs proliferation and inhibited its adipogenic differentiation at 1.0 mmol/L and 5.0 mmol/L, which could be associated the facts that it promoted osteogenic differentiation and maintain the balance between osteoblastogenesis and adipogenesis.

关 键 词：雷奈酸锶 骨髓间充质干细胞 细胞增生 成脂分化 骨质疏松 

分 类 号：R681[医药卫生—骨科学]