唑来膦酸对破骨细胞分化中钙调蛋白、钙调磷酸酶表达的影响
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The effect of zoledronate on gene expression of calmodulin and calcineurin during osteoclast differentiation
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    摘要:

    目的:探讨唑来膦酸(zoledronate,ZOL)对破骨细胞分化中钙调蛋白(calmodulin)和钙调磷酸酶(calcineurin)基因表达的影响-方法:小鼠RAW264.7细胞分为A-B两组,均用100 ng/mL核因子κB受体激活蛋白配体(RANKL)诱导4 d;B组细胞在RANKL 诱导第2天加入1×10-6 mol/L ZOL处理48 h-RANKL 诱导4 d后收获细胞,检测破骨细胞生成及calmodulin-calcineurin基因表达情况-结果:B组多核破骨细胞数-吸收陷窝数目及面积分别为(8.8 ± 2.3)个-(6.7 ± 1.2)个和(997.1 ± 14.8)μm2,均显著低于A组的(19.7 ± 2.9)个-(13.3 ± 1.5)个和(1 676.9 ± 24.9) μm2(P < 0.01)-B组calmodulin mRNA及蛋白水平较A组分别降低了51.0%和78.7%(P < 0.05),calcineurin则分别下降了37.0%和69.5%(P < 0.01)-免疫荧光细胞化学显示,B组calmodulin-calcineurin的荧光强度较A组也明显下降-结论:ZOL可显著抑制破骨细胞生成,并下调破骨细胞分化中calmodulin和calcineurin的mRNA和蛋白水平,而calmodulin-calcineurin可能参与了ZOL对破骨细胞形成和功能的抑制-

    Abstract:

    Objective:The study is aimed to examine the effects of zoledronate(ZOL) on gene expression of calmodulin and calcineurin during differentiation of osteoclasts. Methods:RAW264.7 cells were divided into 2 groups,group A and group B. The cells in both groups were induced with 100 ng/ml RANKL(ligand of receptor activator of nuclear factor κB)for 4 d for osteoclast differentiation. At the second day of RANKL induction,cells in group B also treated with 1×10-6 mol/L ZOL for 48 h. Four days later,the cells were harvested and examined for osteoclastogenesis,as well as gene expression of both calmodulin and calcineurin. Results: TRAP positive multinuclear cells,number and size of resorption lacunaes in group B were 8.8 ± 2.3,6.7 ± 1.2 and 997.1 ± 14.8 μm2 respevtively,which were significantly lower than those (19.7 ± 2.9,13.3 ± 1.5 and 1676.9 ± 24.9 μm2)in group A (P < 0.01). mRNA and protein level of calmodulin in group B decreased about 51% and 78.7%,respectively when compared with group A (P < 0.05),and those of calcineurin also decreased about 37% and 69.5%,respectively(P < 0.05). Immunofluorescent cytochemistry showed obvious decrease of fluorescent intensity of calmodulin and calcineurin protein in group B when compared with group A. Conclusion:ZOL significantly inhibits osteoclastogenesis and down-regulates gene expression of calmodulin and calcineurin,which may play a role in ZOL-induced inhibition of osteoclastogenesis.

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阴露佳,林珏杉,温黎明,董 伟,冯晓洁,孙 红,戚孟春.唑来膦酸对破骨细胞分化中钙调蛋白、钙调磷酸酶表达的影响[J].南京医科大学学报(自然科学版),2016,(2):171-175

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  • 收稿日期:2015-07-23
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  • 在线发布日期: 2016-02-29
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