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作 者:张金超[1] 李亚平[1] 杨康宁[1] 郝晓红[1]
机构地区:[1]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
出 处:《无机化学学报》2010年第12期2251-2258,共8页Chinese Journal of Inorganic Chemistry
基 金:国家自然科学基金(No.20971034);河北省自然科学基金(No.B2009000161);教育部科学技术研究重点项目(No.208018);河北省留学人员科技活动项目;河北大学自然科学基金资助项目
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1×10-5mol.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.
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