摘要
为探讨同型半胱氨酸对人单核细胞表达和分泌巨噬细胞炎性蛋白 1α的影响并检测其活性 ,在体外培养的THP 1单核细胞培养基中加入终浓度为 0 .0 5mmol L、0 .1mmol L和 0 .2mmol L的同型半胱氨酸 ,孵育 8h ,或加入终浓度 0 .1mmol L的同型半胱氨酸 ,分别孵育 4、8及 16h ,用酶联免疫吸附法检测其培养基中的分泌性巨噬细胞炎性蛋白 1α。用改良Boyden小室微孔滤膜法检测THP 1单核细胞条件培养基中巨噬细胞炎性蛋白 1α蛋白对外周血单核细胞的趋化活性。结果发现 ,培养的THP 1单核细胞暴露于不同浓度或同一浓度不同孵育时间的同型半胱氨酸后 ,其分泌至条件培养基中巨噬细胞炎性蛋白 1α蛋白呈浓度和时间依赖性递增 (P <0 .0 1)。趋化实验表明 ,经同型半胱氨酸刺激的THP 1条件培养基引起单核细胞迁移距离 ( 83 .2± 4.8μm)明显大于未经同型半胱氨酸刺激的条件培养基组 ( 73 .2± 2 .3 μm)和随机移动组 ( 62 .4± 3 .5 μm)。方差分析显示 ,三组间均有显著性差异 (F =3 10 .70 ,P <0 .0 1)。在经同型半胱氨酸刺激的THP 1条件培养基中加入巨噬细胞炎性蛋白 1α抗体后 ,单核细胞的移动距离较不含抗体组明显缩短 (P <0 .0 1)。结果提示 ,同型半胱氨酸能诱导THP 1单核细胞表达有趋化活性的分泌性巨噬细胞炎性蛋?
Aim To investigate whether homocysteine (HCY) induce the expression of secretory macrophage inflammatory protein-1α (MIP-1α)in THP-1 monocytes. Methods After exposure of the cultured THP-1 monocytes to HCY at increasing concentrations (0.05, 0.1 and 0.2 mmol/L) for 8 h, or at a concentration of 0.1 mmol/L for different incubation times (4, 8 and 16 h), enzyme-linked immunosorbent assay (ELISA) was used to determine MIP-1α protein expression in the cultured supernate of each group. Conditioned media with or without HCY (0.01 mmol/L) were collected for chemotaxis assay by micropore filter method using a modified Boyden Chamber. Results ELISA showed that the exposure of THP-1 monocytes to HCY at different concentrations and for different times induced higher production of MIP-1α protein compared with the control (P<0.01). The MIP-1α protein in cultured supernate was increased in a dose- and time-dependent manner. In chemotaxis assay, the migration distance of monocytes in HCY stimulating group was 83.2±4.8 μm, which was significantly longer than that in the non-HCY stimulating group (73.2±2.3 μm) and random migration group (62.4±3.5 μm)(F=310.70, P<0.01). After adding goat anti-human MIP-1α antibody, the monocyte chemotactic activity was noticeably inhibited, compared with the HCY stimulating group without specific antibody (P<0.01). Conclusions HCY can induce THP-1 monocytes to express MIP-1αand secret into culture medium, which share partial chemotactic activity for peripheral monocytes, and may play an important role in atherogenesis through promoting the recruitment of monocytes into arterial intima.
出处
《中国动脉硬化杂志》
CAS
CSCD
2003年第1期9-12,共4页
Chinese Journal of Arteriosclerosis
基金
国家自然科学基金 ( 39730 2 2 0 )资助
