摘要
目的探讨17-β雌二醇对大鼠成骨细胞β2肾上腺素受体(β2AdR)表达的影响。方法新生SD大鼠颅骨经多次酶消化、反复贴壁和分离纯化培养得到纯净成骨细胞并进行碱性磷酸酶(ALP)染色。加入不同浓度17-β雌二醇(10-6～10-9mmol/L)培养,在不同时间点采用RT-PCR法检测细胞β2AdR mRNA基因表达,采用Western blot法检测细胞中β2AdR蛋白的表达。结果 RT-PCR和Western blot检测结果表明,随着17-β雌二醇浓度的升高,7、14d培养组成骨细胞β2AdR mRNA和β2AdR蛋白表达水平均逐渐下降,并呈现药物浓度依赖性,对照组表达水平最高。结论 17-β雌二醇可抑制成骨细胞β2AdR的表达,可能是其调控骨代谢的机制之一。
Objective To evaluate the effect of 17β-estradiol on the expression of β2 adrenergic receptor of osteoblast cells.Methods A large amount of pure osteoblast cells collected from cranial bones of female newly born SD rats with many times enzyme digestion and repetitive adhesion method were isolated and cultured.Osteoblast were confirmed by morphology and alkaline phosphatase staining.Different concentrations 17β-estradiol was added into the culture.RT-PCR was used to detect the β2AdR gene expression and protein level of β2 adrenergic receptor were measured by western blot at 7th and 14th days.Results With increased concentration of 17β-estradiol,the level of β2AdR mRNA and β2-adrenergic receptor protein decreased at both time,comparing with the control group.Conclusion 17β-estradiol can inhibit the expressions of β2AdR gene and β2-adrenergic receptor in the rat osteoblast cells in a concentration dependent manner,and that may be one of the regulatory mechanisms of bone metabolism.
出处
《中华骨质疏松和骨矿盐疾病杂志》
2012年第1期35-40,共6页
Chinese Journal Of Osteoporosis And Bone Mineral Research
基金
国家自然科学基金面上项目(30600626)
