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福莫特罗和ICI118551对大鼠骨髓间充质干细胞来源的成骨样细胞增殖和分化的影响

The Effects of Formoterol and ICI118551 on the Proliferation and Differention of Rat Osteoblast-like Cells Induced from Marrow-derived Mesenchymal Stem Cells in Vitro

作者: 专业:骨科学 导师:韦永中 年度:2009 学位:硕士  院校: 南京医科大学

Keywords

Formoterol, ICI118551, Rat marrow-derived mesenchymal stem cells, alkali phosphatase, osteocalcin, proliferation, apoptosis

        [目的]:研究选择性β2肾上腺素能受体激动剂福莫特罗(Formoterol)和选择性β2肾上腺素能受体阻滞剂ICI118551对体外大鼠骨髓间充质干细胞(MMSC)来源的成骨样细胞增殖和分化的影响,探讨β2肾上腺素能受体信号对骨代谢的影响。[方法]:取三周龄雌性SD大鼠,全贴壁筛选法培养MMSC,用条件培养液诱导向成骨细胞分化后分别加入不同浓度Formoterol和ICI118551,硝基苯磷酸二钠(PNPP)法检测细胞碱性磷酸酶,放射免疫法检测骨钙素的分泌水平,噻唑蓝(MTT)法检测细胞增殖,Tunel末端标记法定性检测细胞凋亡,Annexin V-FITC/PI双标法定量检测细胞凋亡率。[结果]:MMSC成功诱导分化为成骨样细胞。同对照组相比,不同浓度(10-5-10-9mol/L)的Formoterol均可抑制成骨样细胞分泌碱性磷酸酶和骨钙素,并可抑制该细胞增殖,但对细胞凋亡无明显影响。同对照组相比,高浓度(10-5和10-6mol/L)的ICI118551抑制成骨样细胞分泌碱性磷酸酶和骨钙素,并可抑制该细胞增殖,低浓度(10-8和10-9 mol/L)的ICI118551则可促进成骨样细胞分泌碱性磷酸酶和骨钙素,并可促进细胞增殖,但是不同浓度(10-5-10-9mol/L)的ICI118551对成骨样细胞的凋亡无明显影响。[结论]:β2肾上腺素能受体激动剂可抑制体外成骨样细胞的增殖和分化;β2肾上腺素能受体阻滞剂对成骨样细胞增殖和分化的影响与其浓度有关;β2肾上腺素能受体激动剂和阻滞剂对成骨样细胞的凋亡无明显影响。
    【Objectives】:To evaluate the effects of Formoterol(β2-adrenergic receptor-specific agonist) and ICI118551(β2-adrenergic receptor-specific antagonist) on rat osteoblast- like cells induced from marrow-derived mesenchymal stem cells(MMSC) .【Methods】:MMSCs by whole bone marrow adherent culture derived from a three-week-old female SD rat are induced to osteoblast-like cells.After different concentrations of Formoterol and ICI118551 are added into the culture medium, the alkali phosphatase is detected by PNPP and the osteocalcin is detected by radio-immunity.The cell proliferation is evaluated with the method of MTT.The cell apoptosis is evaluated by the methods of Annexin V-FITC/PI and Tunel.【Results】: The alkali phosphatase,osteocalcin and proliferation of cells cultured with all different concentrations(10-5-10-9 mol/L) of Formoterol are obviously lower that of the control.As to ICI118551, high concentrations(10-5 and 10-6 mol/L) can inhibit the alkali phosphatase, osteocalcin and proliferation of cells while low concentrations (10-8 and 10-9mol/L) can enhance the alkali phosphatase,osteocalcin and proliferation.Both Formoterol and ICI118551 have minimal effects on the cell apoptosis.【Conclusions】: Theβ2-adrenergic receptor-specific agonist Formoterol can inhibit the activity of rat osteoblast-like cells in vitro while the effects of theβ2-adrenergic receptor-specific antagonist ICI118551 are not single, depending on its concentration.Both of them have minimal effects on the cell apoptosis.
        

福莫特罗和ICI118551对大鼠骨髓间充质干细胞来源的成骨样细胞增殖和分化的影响

缩略语4-5
中文摘要5-7
英文摘要7-8
前言9-10
材料与方法10-20
结果20-38
分析和讨论38-42
小结42-43
参考文献43-47
综述47-61
    参考文献55-61
攻读学位期间发表文章情况61-62
致谢62
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