本文選題：神經(jīng)肽P物質(zhì) + 成骨細(xì)胞��； 參考：《上海交通大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)》2017年01期

【摘要】：目的·探討神經(jīng)肽P物質(zhì)(SP)對(duì)轉(zhuǎn)錄因子RUNX2表達(dá)的調(diào)控及其對(duì)成骨細(xì)胞增殖能力的影響。方法·體外分離C57BL/6J小鼠顱骨來(lái)源的原代成骨細(xì)胞,通過(guò)轉(zhuǎn)染si RNA沉默成骨細(xì)胞RUNX2基因的表達(dá)。將SP、Spantide(SP抑制劑)、L703606(NK-1受體抑制劑)以及RUNX2 si RNA進(jìn)行配伍作用于成骨細(xì)胞,觀察SP對(duì)成骨細(xì)胞RUNX2表達(dá)的調(diào)控及對(duì)成骨細(xì)胞增殖能力的影響。成骨細(xì)胞增殖活性的檢測(cè)采用細(xì)胞增殖-毒性檢測(cè)試劑盒(CCK-8法)。通過(guò)real-time PCR和Western blotting技術(shù)檢測(cè)SP對(duì)RUNX2在m RNA和蛋白表達(dá)水平的調(diào)控。結(jié)果·CCK-8法檢測(cè)結(jié)果顯示,濃度為10~(-12)~10~(-6) mol/L的SP促進(jìn)小鼠成骨細(xì)胞增殖,10~(-8) mol/L為其最佳濃度。10~(-8) mol/L SP作用于成骨細(xì)胞48 h后,轉(zhuǎn)錄因子RUNX2的m RNA和蛋白表達(dá)水平較對(duì)照組明顯升高,Spantide和L703606抑制SP對(duì)RUNX2表達(dá)的誘導(dǎo)。RUNX2 si RNA降低成骨細(xì)胞基礎(chǔ)增殖水平,同時(shí)也抑制了SP對(duì)成骨細(xì)胞的促增殖作用。結(jié)論·SP通過(guò)與NK-1受體結(jié)合誘導(dǎo)轉(zhuǎn)錄因子RUNX2的表達(dá),進(jìn)而增強(qiáng)成骨細(xì)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.

【作者單位】： 上海交通大學(xué)醫(yī)學(xué)院附屬新華醫(yī)院骨科;
【分類號(hào)】：R336

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