Cu^(2+)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 Cu^(2+) and Cu^+ on the Proliferation,Differentiation and Calcification of Primary Mouse Osteoblasts in vitro

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1×10-5mol.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,1×10-5mol.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。

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.