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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 effect of homocysteine (Hcy) on the expression of monocyte chemoattractant protein 1 (MCP 1) in macrophages (THP 1) differentiated from human monocytic (THP 1) Regulatory effect of statins. Methods: The cell density was adjusted to 1.0 × 10 6 cells / ml. After the medium was added with PMA, the final concentration was 20 ng / ml. After cultured for 48 h, the suspension cells were discarded and the adherent cells were macrophage-like Differentiation. The statin group and the Hcy group were incubated with simvastatin (10 μmol / L, 50 μmol / L) or without simvastatin for 24 h, and then completely cultured with DL Hcy (0.1 mmol / L) The control group only with complete medium, cultured at 37 ℃ 2 ~ 2 4h, the supernatant collected 500g centrifuged 10min, -20 ℃ preservation. The amount of MCP 1 protein in the supernatant was measured by ELISA and was repeated 3 times per well. Results: Compared with the control group, the MCP1 protein expression in THP1 macrophages increased at 0 .1 mmol / LHcy and significantly increased at 4 h (P <0.05), reaching the peak at 6 h (P <0. 0 1), and then decreased gradually after 12 hours (P <0.05), which were 2.9 times, 5.8 times and 2.6 times higher than that of the control group respectively. There was no significant difference between the control group and the 24th hour. Simvastatin at 10 μmol / L and 50 μmol / L, respectively, decreased the MCP 1 protein level of 6 h peak when incubated with Hcy to 57.11% and 23.6% of the statin group, respectively. CONCLUSION: Hcy with pathological concentration (0.1 mmol / L) can promote the MCP 1 protein expression in THP 1 macrophages in a time-dependent manner, which can be down-regulated by simvastatin.