羧甲基赖氨酸对人外周血单核细胞破骨样转化的影响-东南大学学报医学版

羧甲基赖氨酸对人外周血单核细胞破骨样转化的影响

单位：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

分类号：R543

出版年·卷·期（页码）：2014·33·第三期（283-287）

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κ B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 N^ε-(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.

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