神经肽P物质通过诱导RUNX2表达对小鼠成骨细胞增殖能力的影响

发布时间：2018-04-26 10:12

本文选题：神经肽P物质 + 成骨细胞　；参考：《上海交通大学学报(医学版)》2017年01期

【摘要】：目的·探讨神经肽P物质(SP)对转录因子RUNX2表达的调控及其对成骨细胞增殖能力的影响。方法·体外分离C57BL/6J小鼠颅骨来源的原代成骨细胞,通过转染si RNA沉默成骨细胞RUNX2基因的表达。将SP、Spantide(SP抑制剂)、L703606(NK-1受体抑制剂)以及RUNX2 si RNA进行配伍作用于成骨细胞,观察SP对成骨细胞RUNX2表达的调控及对成骨细胞增殖能力的影响。成骨细胞增殖活性的检测采用细胞增殖-毒性检测试剂盒(CCK-8法)。通过real-time PCR和Western blotting技术检测SP对RUNX2在m RNA和蛋白表达水平的调控。结果·CCK-8法检测结果显示,浓度为10~(-12)~10~(-6) mol/L的SP促进小鼠成骨细胞增殖,10~(-8) mol/L为其最佳浓度。10~(-8) mol/L SP作用于成骨细胞48 h后,转录因子RUNX2的m RNA和蛋白表达水平较对照组明显升高,Spantide和L703606抑制SP对RUNX2表达的诱导。RUNX2 si RNA降低成骨细胞基础增殖水平,同时也抑制了SP对成骨细胞的促增殖作用。结论·SP通过与NK-1受体结合诱导转录因子RUNX2的表达,进而增强成骨细胞增殖能力。
[Abstract]:Aim to investigate the regulation of neuropeptide P (SP) on the expression of transcription factor RUNX2 and its effect on the proliferation of osteoblasts. Methods: isolation of primary osteoblasts from C57BL/6J mice in vitro, and the expression of RUNX2 gene in osteoblasts by transfection of Si RNA. SP, Spantide (SP inhibitor), L703606 (NK-1 receptor inhibitors), and the expression of RUNX2 gene in osteoblasts were transfected in vitro. RUNX2 Si RNA was compatible with osteoblasts, observed the regulation of SP on the expression of RUNX2 in osteoblasts and the effect on the proliferation of osteoblasts. The proliferation activity of osteoblasts was detected by cell proliferation toxicity detection kit (CCK-8 method). Real-time PCR and Western blotting techniques were used to detect SP on RUNX2 and protein expression water. Results. Results. The results of CCK-8 assay showed that the SP of 10~ (-12) ~10~ (-6) mol/L promoted the proliferation of osteoblasts in mice. 10~ (-8) mol/L was the best concentration.10~ (-8), and the expression level of transcription factor and protein expression level were significantly higher than that of the control group. The expression of induced.RUNX2 Si RNA reduces the proliferation level of osteoblast base and inhibits the proliferation promoting effect of SP on osteoblasts. Conclusion: SP can induce the expression of transcription factor RUNX2 by binding to NK-1 receptor, and then enhance the proliferation of osteoblasts.

【作者单位】：上海交通大学医学院附属新华医院骨科;
【分类号】：R336

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