Comparative effects of five bisphosphonates on apoptosis of macrophage cells in vitro
Comparative effects of five bisphosphonates on apoptosis of macrophage cells in vitro
复制标题
DOI：
10.1016/j.bcp.2006.09.031
复制
发表时间：
2007-03-01
影响因子：
5.8
通讯作者：
Chappard, D.
中科院分区：
文献类型：
--
作者：
Moreau, M. F.;Guillet, C.;Chappard, D.
双磷酸盐（BPs）通过降低骨细胞活性抑制骨吸收;它们诱导破骨细胞凋亡。假体松动的病理生理学是复杂的，意味着继发于巨噬细胞吞噬磨损碎屑的炎症反应，继发于破骨细胞增加的骨吸收。BPs通过诱导细胞凋亡抑制巨噬细胞的增殖并导致细胞死亡。我们使用小鼠巨噬细胞样J774.1细胞来评价五种BP的作用。J774 A. 1细胞在标准培养基中培养2天。在3天内将BP（阿仑膦酸盐、帕米膦酸盐、依替膦酸盐、利塞膦酸盐、唑来膦酸）以10（-6）至10（-4）M的浓度加入培养基中。在用荧光染料Hoescht H33342染色后通过荧光显微镜研究细胞，并在300个细胞核上测定凋亡细胞的百分比。在用膜联蛋白V-FITC（用于计数凋亡细胞）和碘化丙啶（用于坏死/晚期凋亡细胞）对2000个细胞染色后，通过流式细胞荧光测定法分析细胞。阿仑膦酸盐和帕米膦酸盐仅在非常高的浓度[10（-4）M]下引起细胞凋亡和死亡。相反，细胞凋亡和坏死的细胞证明与利塞膦酸盐或唑来膦酸在较低的浓度。这些效应具有剂量依赖性，并在浓度达到[10（-5）M]时发生。唑来膦酸组凋亡细胞数高于利塞膦酸组。在巨噬细胞凋亡的研究中，细胞荧光测定法似乎上级于细胞学分析，因为坏死细胞与载玻片松散接触，并且在细胞学计数中无法识别。一些氨基-BP似乎诱导巨噬细胞凋亡。(c)2006年爱思唯尔公司All rights reserved.
Bisphosphonates (BPs) inhibits bone resorption by reducing osteoclastic activity; they induce osteoclast apoptosis. Pathophysiology of prostheses loosening is complex and implies an inflammatory reaction secondary to the phagocytosis of wear debris by macrophages with a secondary increased bone resorption by osteoclasts. BPs inhibit proliferation and cause cell death in macrophages by induction of apoptosis. We have used mouse macrophage-like J774.1 cells to evaluate the effects of five BPs.J774A.1 cells were cultured in a standard culture medium for 2-days. BPs (alendronate, pamidronate, etidronate, risedronate, zoledronic acid) were added in the medium at concentration of 10(-6) to 10(-4) M during 3 days. Cells were studied by fluorescence microscopy after staining with the fluorescent dye Hoescht H33342 and the percentage of apoptotic cells was determined on 300 nuclei. Cells were analyzed by flow cytofluorometry after staining with annexin V-FITC (for counting apoptotic cells) and propidium iodide (for necrotic/late-apoptotic cells) on 2000 cells.Etidronate did not cause significant apoptosis or necrosis, at any concentration. Alendronate and pamidronate caused apoptosis and death only at very high concentration [10(-4) M]. On the contrary, apoptotic and necrotic cells were evidenced with risedronate or zoledronic acid at lower concentrations. These effects were dose-dependant and occurred when concentration reached [10(-5) M]. The number of apoptotic cells was higher with zoledronic acid and then with risedronate. Cytofluorometry appeared superior to cytologic analysis in the investigation of macrophage apoptosis, since necrotic cells loose contact with the glass slides and are not identifiable in cytological counts. Some amino-BPs appear to induce apoptosis in macrophages. (c) 2006 Elsevier Inc. All rights reserved.