【摘要】：目的研究活化T細(xì)胞核因子蛋白c1(NFATc1)過(guò)表達(dá)對(duì)唑來(lái)膦酸誘發(fā)的破骨細(xì)胞生成抑制的挽救效應(yīng)。方法小鼠NFATc1重組慢病毒轉(zhuǎn)染RAW264.7細(xì)胞,驗(yàn)證NFATc1蛋白表達(dá)。細(xì)胞分為A、B兩組,分別轉(zhuǎn)染空白載體和NFATc1重組載體。兩組細(xì)胞均用50 ng/mL核因子κB受體激活蛋白配體(RANKL)誘導(dǎo)5 d,并在第2天加入1×10~(-6)mmol/mL唑來(lái)膦酸作用2 d。第6天收獲細(xì)胞,檢測(cè)破骨細(xì)胞的生成和功能及NFATc1、抗酒石酸酸性磷酸酶(TRAP)、酪氨酸激酶(c-Src)基因的表達(dá)情況。結(jié)果 NFATc1蛋白在細(xì)胞中均有表達(dá)。B組TRAP陽(yáng)性破骨細(xì)胞數(shù)、牙本質(zhì)吸收陷窩數(shù)目和面積顯著高于A組(P0.01)。B組NFATc1、TRAP、c-Src的蛋白水平顯著高于A組(P0.01);B組3個(gè)基因mRNA水平顯著高于A組(P0.01)。免疫熒光細(xì)胞化學(xué)檢測(cè)也得到相似的結(jié)果。結(jié)論 NFATc1過(guò)表達(dá)對(duì)唑來(lái)膦酸誘發(fā)的破骨細(xì)胞生成及NFATc1、TRAP、c-Src基因表達(dá)的抑制具有挽救效應(yīng);NFATc1作為Ca~(2+)信號(hào)的關(guān)鍵分子,在唑來(lái)膦酸誘發(fā)的破骨細(xì)胞生成抑制中起著關(guān)鍵作用。
文內(nèi)圖片：
圖片說(shuō)明：Westernblot檢測(cè)ＲAW264．7細(xì)胞內(nèi)NFATc1蛋白表達(dá)2．2破骨細(xì)胞生成及骨吸收功能檢測(cè)經(jīng)TＲAP
[Abstract]:Objective to study the effect of overexpression of activated T nuclear factor protein C1 (NFATc1) on osteoclast formation induced by zoledronic acid. Methods RAW264.7 cells were transfected with mouse NFATc1 recombinant lentivirus and the expression of NFATc1 protein was verified. The cells were divided into two groups: group A and group B, and were transfected with blank vector and NFATc1 recombinant vector, respectively. Both groups were induced with 50 ng/mL nuclear factor kappa B receptor activating protein ligand (RANKL) for 5 days, and 1 脳 10 ~ (- 6) mmol/mL zoledronic acid was added on the 2nd day for 2 d. On the 6th day, the formation and function of osteoclasts and the expression of NFATc1, anti-tartrate acid phosphatase (TRAP), tyrosine kinase (c-Src) gene were detected. Results NFATc1 protein was expressed in all the cells. The number of TRAP positive osteoclasts, the number and area of dentin resorption lacunae in group B were significantly higher than those in group A (P 0.01, the protein level of NFATc1,TRAP,c-Src in group). B was significantly higher than that in group A (P 0.01), and the level of mRNA of three genes in group B was significantly higher than that in group A (P 0.01). Similar results were obtained by immunofluorescence cytochemistry. Conclusion NFATc1 overexpression has a rescue effect on osteoclast formation and NFATc1,TRAP,c-Src gene expression induced by zoledronic acid. NFATc1, as a key molecule of Ca~ (2) signal, plays a key role in osteoclast formation inhibition induced by zoledronic acid.
【作者單位】： 華北理工大學(xué)口腔醫(yī)學(xué)院口腔頜面外科教研室;華北理工大學(xué)基礎(chǔ)醫(yī)學(xué)院病理教研室;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(編號(hào):81270965) 河北省高等學(xué)�？茖W(xué)技術(shù)研究重點(diǎn)項(xiàng)目(編號(hào):ZD2015005) 河北省自然科學(xué)基金資助項(xiàng)目(編號(hào):2011401044)
【分類號(hào)】：R965

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