降钙素对大鼠颅骨成骨细胞调控作用的体外观察

发布时间：2018-02-23 22:33

本文关键词： 骨折降钙素成骨细胞分化蛋白激酶类转录因子细胞外信号调节激酶核心结合因子促分裂原活化蛋白激酶信号通路抑制剂　出处：《中国组织工程研究与临床康复》2009年02期 　论文类型：期刊论文

【摘要】：背景:降钙素是强有力的破骨细胞抑制剂,可直接、快速而广泛地抑制骨吸收,近年来有学者报道其有促进骨折愈合的功能,推测降钙素对成骨细胞也有直接的作用。目的:体外观察降钙素对大鼠成骨细胞的药理作用,分析其是否通过细胞外信号调节激酶信号通路产生。设计、时间及地点:分组对照观察,于2007-06/2008-05在南京医科大学药理学实验室完成。材料:选用新生SD大鼠,分离培养颅骨成骨细胞。方法:取第2代成骨细胞进行分组实验:0.01μg/L降钙素组、0.1μg/L降钙素组、1μg/L降钙素组分别加入不同剂量降钙素进行干预,U0126+1μg/L降钙素组提前30min加入100μmol/L的细胞外信号调节激酶抑制剂U0126,并设空白对照组。主要观察指标:通过四甲基偶氮唑盐比色法、对硝基苯磷酸盐法和钙结节茜素红染色法分别检测降钙素对成骨细胞的增殖、分化、矿化能力的影响;应用反转录-聚合酶链反应技术检测促骨形成关键基因核心结合因子a1mRNA的表达情况;应用WesternBlotting技术检测磷酸化细胞外信号调节激酶1/2蛋白表达情况。结果:加入0.01,0.1,1μg/L的降钙素干预后,成骨细胞的增殖、分化、矿化能力以及核心结合因子a1mRNA的表达均随降钙素剂量增加而增强,且0.1,1μg/L组与空白对照组比较差异具有显著意义(P0.05～0.01);此外降钙素促进了磷酸化细胞外信号调节激酶1/2蛋白的表达(P0.05)。U0126可阻断以上所有效应(P0.05～0.01)。结论:降钙素可能通过细胞外信号调节激酶信号通路调控促骨形成关键基因核心结合因子a1mRNA的表达,进而促进成骨细胞的增殖、分化、矿化能力。
[Abstract]:Background: calcitonin is a powerful osteoclast inhibitor that can inhibit bone resorption directly, quickly and extensively. In recent years, it has been reported that calcitonin can promote fracture healing. Objective: to observe the pharmacological effect of calcitonin on rat osteoblasts in vitro and analyze whether calcitonin regulates the production of kinase signal pathway by extracellular signal. Time and place: group controlled observation, completed in the Laboratory of Pharmacology, Nanjing Medical University, 2007-06 / 2008-05. Materials: newborn SD rats, Methods: the osteoblasts of the second passage of calcitonin group were divided into 0. 1 渭 g / L calcitonin group and 1 渭 g / L calcitonin group. U0126, an extracellular signal-regulated kinase inhibitor, and set up a blank control group. Main outcome measures: tetramethylazolium colorimetry, The effects of calcitonin on the proliferation, differentiation and mineralization of osteoblasts were detected by p-nitrophenyl phosphate method and alizarin red staining. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of core binding factor A1 mRNA. WesternBlotting technique was used to detect the expression of extracellular signal-regulated kinase 1/2. Results: osteoblasts proliferated and differentiated after the intervention of 0.01 渭 g / L calcitonin. The mineralization ability and the expression of core binding factor A1 mRNA increased with the increase of calcitonin dose. In addition, calcitonin promoted the expression of extracellular signal-regulated kinase 1/2 protein (P0.05N. U0126). Conclusion: calcitonin may pass through the extracellular pathway. Signal regulated kinase signaling pathway regulates the expression of core binding factor A1 mRNA, which is a key gene for bone formation. And then promote the proliferation, differentiation and mineralization of osteoblasts.
【作者单位】：南京医科大学附属南京儿童医院;
【分类号】：R96

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