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羧甲基赖氨酸对人外周血单核细胞破骨样转化的影响 预览 被引量:4

Effects of N ^ε- (Carboxymethyl)lysine on human peripheral blood mononcytes differentiation to osteoclast-like cells
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摘要 目的:探讨羧甲基赖氨酸( CML)对外周血单核细胞破骨样转化的影响。方法:从外周血分离提取单核细胞,分为3组:细胞核因子κB受体活化因子配基( RANK-L)+巨噬细胞集落刺激因子( M-CSF)诱导组( A组),CML+RANK-L+M-CSF干预组( B组),空白对照组( C组)。利用RANK-L和M-CSF诱导其破骨样转化,每天通过倒置显微镜观察细胞生长形态及计算细胞核个数,培养8d后通过抗酒石酸酸性磷酸酶(TRAP)染色法观察破骨样细胞的形成,Western blotting 检测破骨细胞标志物组织蛋白酶K蛋白表达。结果:随着培养时间的延长,A组TRAP阳性多核细胞数明显增加,B组TRAP阳性多核细胞数较A组明显减少(P<0.05),B组组织蛋白酶K表达亦较A组明显降低(P<0.05)。结论:CML能够抑制外周血单核细胞向破骨样细胞转化。 Objective:To investigate the effects of Nε-(Carboxymethyl)lysine(CML)on osteoclast-like differenti-ation in human peripheral blood monocytes (PBMCs).Methods:PBMCs were isolated from blood of healthy donors by centrifugation on Ficoll-Hypaque Plus .Cells were inducted differentiation into osteoclasts in the presence of receptor activator of nuclear factor kappa-B ligand(RANKL) and macrophage colony stimulating factor (M-CSF). After 8 d culture, morphologic characteristics and multinucleated osteoclast-like cell formation were observed through inverted microscope .Tartrate-resistant acid phosphatase ( TRAP ) positive cells were stained by using a TRAP staining kit . Cathepsin K protein expression was examined by Western blotting . Results: With the prolongation of the culture time , a great amount of multinucleated and TRAP positive cells could be found in induction group . Multinucleated and TRAP positive cells decreased significantly in CML treatment group . Conclusion:CML suppressed osteoclast-like differentiation in human PBMCs .
作者 魏芹 金虹 任晓妹 蒋益波 刘乃丰 WEI Qin, JIN Hong, REN Xiao-mei, JIANG Yi-bo, LIU Nai-feng ( 1. College of Medicine, Southeast Universtity, Nanjing 210009, China ; 2. Department of Cardiology, Zhongda Hospital, Southeast Universtity, Nanjing 210009, China; 3. Department of Cardiology, Ren Min Hospital of Taixing, Taixing 225400, China)
出处 《东南大学学报:医学版》 CAS 2014年第3期283-287,共5页 Journal of Southeast Univ: Medical Sci Ed
基金 国家自然科学基金资助项目(81170254,81300227) 江苏省社会发展基金资助项目(BE2011793)
关键词 羧甲基赖氨酸 单核细胞 破骨样细胞 分化 N^ε- (Carboxymethyl) lysine mononcyte osteoclast-like cell differentiation
作者简介 魏芹(1978-),女,江苏铜山人,主治医师,医学硕士,在读博士研究生。E—mail:weiqin1102@163.com [通信作者]刘乃丰E—mail:liunf_2006@126.com
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