本文選題：神經(jīng)肽P物質(zhì) + 成骨細胞��； 參考：《上海交通大學學報(醫(yī)學版)》2017年01期

【摘要】：目的·探討神經(jīng)肽P物質(zhì)(SP)對轉(zhuǎn)錄因子RUNX2表達的調(diào)控及其對成骨細胞增殖能力的影響。方法·體外分離C57BL/6J小鼠顱骨來源的原代成骨細胞,通過轉(zhuǎn)染si RNA沉默成骨細胞RUNX2基因的表達。將SP、Spantide(SP抑制劑)、L703606(NK-1受體抑制劑)以及RUNX2 si RNA進行配伍作用于成骨細胞,觀察SP對成骨細胞RUNX2表達的調(diào)控及對成骨細胞增殖能力的影響。成骨細胞增殖活性的檢測采用細胞增殖-毒性檢測試劑盒(CCK-8法)。通過real-time PCR和Western blotting技術(shù)檢測SP對RUNX2在m RNA和蛋白表達水平的調(diào)控。結(jié)果·CCK-8法檢測結(jié)果顯示,濃度為10~(-12)~10~(-6) mol/L的SP促進小鼠成骨細胞增殖,10~(-8) mol/L為其最佳濃度。10~(-8) mol/L SP作用于成骨細胞48 h后,轉(zhuǎn)錄因子RUNX2的m RNA和蛋白表達水平較對照組明顯升高,Spantide和L703606抑制SP對RUNX2表達的誘導。RUNX2 si RNA降低成骨細胞基礎(chǔ)增殖水平,同時也抑制了SP對成骨細胞的促增殖作用。結(jié)論·SP通過與NK-1受體結(jié)合誘導轉(zhuǎn)錄因子RUNX2的表達,進而增強成骨細胞增殖能力。
[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】： 上海交通大學醫(yī)學院附屬新華醫(yī)院骨科;
【分類號】：R336

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