Cu2+和Cu+对原代培养的小鼠成骨细胞增殖、分化和钙化的影响(英文)
Effects of Cu2+ and Cu+ on the Proliferation, Differentiation and Calcification of Primary Mouse Osteoblasts in vitro
作者单位
张金超 河北大学化学与环境科学学院河北省化学生物学重点实验室保定 071002 
李亚平 河北大学化学与环境科学学院河北省化学生物学重点实验室保定 071002 
杨康宁 河北大学化学与环境科学学院河北省化学生物学重点实验室保定 071002 
郝晓红 河北大学化学与环境科学学院河北省化学生物学重点实验室保定 071002 
摘要: 利用噻唑蓝(MTT)法、碱性磷酸酶(ALP)比活性测定、油红O染色和矿化结节染色及定量分析,研究了Cu2+和Cu+对原代培养的成骨细胞增殖、分化及钙化的影响。结果显示:Cu2+(1×10-9~1×10-6 mol·L-1)促进成骨细胞增殖,随时间延长,促进作用变弱。Cu+(1×10-7~1×10-5 mol·L-1)抑制成骨细胞增殖,随时间延长,浓度为1×10-6 mol·L-1的Cu+为促进作用,其余浓度则没有影响。对于成骨细胞分化,Cu2+和Cu+表现出相似的影响,浓度为1×10-9和1×10-6 mol·L-1时均促进成骨细胞分化,而当浓度为1×10-7和1×10-5 mol·L-1时,则抑制成骨细胞分化,随作用时间延长,大多数浓度均表现为促进作用。测试浓度下的Cu2+和Cu+均对成骨细胞向脂肪细胞的横向分化表现为促进效应。对矿化功能的影响,1×10-5 mol·L-1的Cu2+和Cu+表现出显著的抑制效应,但随浓度降低,抑制效应变弱。1×10-7 mol·L-1的Cu2+ 促进成骨细胞矿化结节的形成。结果提示:作用浓度、作用时间及铜离子的价态都是影响Cu2+和Cu+生物效应转变(从毒性到活性,从损伤到保护,从下调到上调)的关键因素。
关键词:   成骨细胞  增殖  分化  钙化
基金项目: 
Abstract: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), alkaline phosphatase (ALP) activity, oil red O assays and alizarin red-S (ARS) stain were employed to evaluate the effects of Cu2+ and Cu+ on proliferation, differentiation and calcification of primary osteoblasts (OBs) in vitro. The results showed that Cu2+ (1×10-9~1×10-6 mol·L-1) promoted the proliferation of OBs for 48 h, and the promotive effect turned to weaken with prolonging incubation time. Cu+(1×10-7~1×10-5 mol·L-1) inhibited the proliferation of OBs for 48 h, but turned to promote the proliferation of OBs at a concentration of 1×10-6 mol·L-1, had no effect on proliferation of OBs at other concentrations for 72 h. Both Cu2+ and Cu+ had similar effects on the differentiation of OBs, they promoted the differentiation of OBs at concentrations of 1×10-9 and 1×10-6 mol·L-1 and inhibited the differentiation at concentrations of 1×10-7 and 1×10-5 mol·L-1 for 48 h, promoted the differentiation at most concentrations for 72 h. Both Cu2+ and Cu+ promoted the adipocytic transdifferentiation of OBs at all tested concentrations. Both Cu2+ and Cu+ (1×10-5 mol·L-1) obviously inhibited the formation of mineralized matrix nodules of OBs, the inhibitory effect turned to weaken with decreasing concentration, Cu2+ promoted the formation of mineralized matrix nodules of OBs at a concentration of 1×10-7 mol·L-1. The results suggest that concentration, culture time and valence state of copper ion are key factors for switching the biological effects of Cu2+ and Cu+ from toxicity to activity, from damage to protection, or from down-regulation to up-regulation.
Keywords: copper  osteoblasts  proliferation  differentiation  calcification
 
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张金超,李亚平,杨康宁,郝晓红.Cu2+和Cu+对原代培养的小鼠成骨细胞增殖、分化和钙化的影响(英文)[J].无机化学学报,2010,26(12):2251-2258.
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