本文關(guān)鍵詞： Klotho 破骨細(xì)胞 RAW.細(xì)胞 抗酒石酸酸性磷酸酶　出處：《細(xì)胞與分子免疫學(xué)雜志》2017年06期 　論文類型：期刊論文

【摘要】：目的探討過表達(dá)Klotho(KL)對可溶性核因子κB受體激活蛋白配體(sRANKL)誘導(dǎo)小鼠RAW264.7細(xì)胞向破骨細(xì)胞分化的影響。方法實(shí)驗(yàn)分為過表達(dá)KL組(Ad-KL組)、空載體組(Ad-GFP組)與空白對照組。倒置熒光顯微鏡觀察重組腺病毒轉(zhuǎn)染RAW264.7細(xì)胞情況,實(shí)時(shí)熒光定量(qRT-PCR)和Western blot法檢測KL的mRNA或蛋白水平;抗酒石酸酸性磷酸酶(TRAP)染色檢測過表達(dá)KL對sRANKL誘導(dǎo)RAW264.7細(xì)胞向破骨細(xì)胞分化的形態(tài)學(xué)影響;qRT-PCR和Western blot法檢測各組破骨細(xì)胞成熟標(biāo)志物TRAP、組織蛋白酶K(CTSK)的mRNA或蛋白水平。結(jié)果與Ad-GFP組和空白對照組比較,Ad-KL組細(xì)胞KL mRNA和蛋白水平顯著升高;TRAP染色顯示,Ad-KL組TRAP陽性細(xì)胞數(shù)量顯少于Ad-GFP組和空白對照組,體積也更小;與Ad-GFP組和空白對照組比較,Ad-KL組TRAP和CTSK mRNA和蛋白水平也明顯降低。結(jié)論過表達(dá)KL抑制RAW264.7細(xì)胞向破骨細(xì)胞分化。
[Abstract]:Objective to investigate the effect of overexpression of Klothotrol (KL) on the differentiation of RAW264.7 cells into osteoclasts induced by soluble nuclear factor 魏 B receptor activator ligand (sRANKL). Methods the cells were divided into two groups: overexpression KL group (Ad-KL group), empty vector group (Ad-GFP group) and blank control group (control group). The transfection of recombinant adenovirus into RAW264.7 cells was observed by inverted fluorescence microscope. The mRNA or protein levels of KL were detected by real-time fluorescence quantitative analysis (qRT-PCR) and Western blot. Morphological effects of overexpression of KL on differentiation of RAW264.7 cells into osteoclasts induced by sRANKL by tartrate-resistant acid phosphatase assay and Western blot assay for detection of mRNA or protein levels of osteoclast maturation marker TRAPK. Results compared with Ad-GFP group and blank control group, the levels of KL mRNA and protein in Ad-KL group were significantly higher than those in Ad-KL group. The results of trap staining showed that the number of TRAP positive cells in Ad-KL group was significantly lower than that in Ad-GFP group and blank control group. Compared with Ad-GFP group and blank control group, the levels of TRAP and CTSK mRNA and protein in Ad-KL group were significantly decreased. Conclusion overexpression of KL inhibits the differentiation of RAW264.7 cells into osteoclasts.
【作者單位】： 重慶醫(yī)科大學(xué)附屬第一醫(yī)院老年病科;重慶市急救醫(yī)療中心老年科;
【基金】：國家自然科學(xué)基金(30672212) 重慶市醫(yī)學(xué)科研計(jì)劃項(xiàng)目(2013-2-029) 重慶市衛(wèi)生計(jì)生委醫(yī)學(xué)科研項(xiàng)目(2016MSXM080) 重慶市渝中區(qū)科技計(jì)劃項(xiàng)目(20150139) 國家臨床重點(diǎn)�？平ㄔO(shè)項(xiàng)目[國衛(wèi)辦醫(yī)函[2013]544號]
【分類號】：R580

【參考文獻(xiàn)】

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