羧甲基赖氨酸对人外周血单核细胞破骨样转化的影响  

Effects of N ^ε- （Carboxymethyl）lysine on human peripheral blood mononcytes differentiation to osteoclast-like cells

作　　者：魏芹[1,2] 金虹[2] 任晓妹[2] 蒋益波[3] 刘乃丰[1,2] 

机构地区：[1]东南大学医学院,江苏南京210009 [2]东南大学附属中大医院心内科,江苏南京210009 [3]泰兴市人民医院心内科,江苏泰兴225400

出　　处：《东南大学学报（医学版）》2014年第3期283-287,共5页Journal of Southeast University(Medical Science Edition)

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)

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8 d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 .

关 键 词：羧甲基赖氨酸 单核细胞 破骨样细胞 分化 

分 类 号：R543[医药卫生—心血管疾病]