摘要
目的 :探讨同型半胱氨酸 (Hcy)促进人单核细胞系 (THP 1)分化而来的巨噬细胞 (THP 1巨噬细胞 )表达单核细胞趋化蛋白 1(MCP 1)及辛伐他汀的调节作用。方法 :调细胞密度到 1.0× 10 6个 /ml,培养液中加入佛波脂 (PMA) ,终浓度为 2 0ng/ml,培养 4 8h ,弃去悬浮细胞 ,贴壁细胞呈巨噬细胞样分化。他汀组和Hcy组分别加入含有辛伐他汀 (10 μmol/L ,5 0 μmol/L)或不含辛伐他汀完全培养基培养 2 4h ,再加入含DL Hcy(0 .1mmol/L)完全培养基 ,对照组只用完全培养基 ,37℃培养 2～ 2 4h ,收集上清 5 0 0g离心 10min ,- 2 0℃保存。用ELISA法检测上清中MCP 1蛋白的量 ,每孔重复 3次。结果 :与对照组比较 ,加入 0 .1mmol/LHcy可使THP 1巨噬细胞MCP 1蛋白表达升高 ,4h后显著升高 (P <0 .0 5 ) ,6h达高峰 (P <0 .0 1) ,12h后逐渐下降 (P <0 .0 5 ) ,分别为对照组的 2 .9倍、5 .8倍和 2 .6倍 ,2 4h与对照组比较差异无显著性意义。 10 μmol/L、5 0 μmol/L辛伐他汀分别使与Hcy共孵育 6h峰值时的MCP 1蛋白量下降至他汀组的5 7.11%、2 3.6 4 %。结论 :病理浓度(0 .1mmol/L)的Hcy可时间依赖性促进THP 1巨噬细胞表达MCP 1蛋白 ,该作用可被辛伐他汀下调。
Objective:To investigate the inductive effect of homocysteine(Hcy) on MCP 1 protein expression in THP 1 derived macrophages (THP 1 macrophage) and regulatory effect of simvastatin. Methods:Human monocytic cells(THP 1) were induced differentiation into macrophages in the present of phorbol 12 myristate 13 acetate(PMA) for 48 hours. Then THP 1 macrophages were incubated in the present or absent of simvastatin prior to Hcy for 24 hours. After washed with 1×PBS slightly, cells were further cultured with Hcy for 2 to 24 hours, while the control were in absent of Hcy. At the end of incubation,the supernatants were harvested. The amount of MCP 1 protein in the supernatants was determined by ELISA. Results:The amount of MCP 1 protein obviously increased at 4, 6, 12 hours after addition of Hcy (P< 0.05 , P< 0.01 ,P< 0.05 ), but decreased to basal level within 24 hours.Pretreatment with simvastatin caused a dose dependent inhibition of MCP 1 protein production in THP 1 macrophages exposed to Hcy. Conclusion:Hcy, at pathological concentration, induces MCP 1 protein expression in THP 1 macrophages, and simvastatin inhibits this effect.
出处
《临床心血管病杂志》
CAS
CSCD
北大核心
2003年第2期109-111,共3页
Journal of Clinical Cardiology
