【摘要】 目的 :探讨同型半胱氨酸是否诱导培养的人脐静脉内皮细胞表达巨噬细胞炎性蛋白 1α。方法 :在内皮细胞培养基中分别加入终浓度为 0 1mmol/L、0 5mmol/L、和 1mmol/L的同型半胱氨酸 ,孵育 8h ,用原位杂交法检测其巨噬细胞炎性蛋白 1αmRNA的表达 ,用细胞酶联免疫吸附实验 (ELISA)检测其巨噬细胞炎性蛋白 1α蛋白的表达。结果 :原位杂交显示 ,培养的人脐静脉内皮细胞能低水平表达巨噬细胞炎性蛋白 1αmRNA ,为胞质内紫蓝色颗粒。培养的内皮细胞暴露于梯度浓度的同型半胱氨酸后其胞质内的巨噬细胞炎性蛋白 1αmRNA明显增加 ,并呈剂量依赖关系 ,组间差异极为显著 (P <0 0 1) ;细胞ELISA显示 ,随着同型半胱氨酸浓度的升高 ,各实验组内皮细胞巨噬细胞炎性蛋白 1α蛋白的表达逐步升高 ,组间差异亦极显著 (P <0 0 1)。结论 :同型半胱氨酸能诱导内皮细胞表达巨噬细胞炎性蛋白 1αmRNA和蛋白 ,可能在单核细胞向动脉内膜的募集中起重要作用。更多还原

【Abstract】 AIM: To investigate the effect of homocysteine (HCY) on the induction of macrophage inflammatory protein-1α (MIP-1α) expression in cultured human umbilical vein endothelial cells (HUVECs). METHODS: After exposure of the cultured HUVECs to HCY at increasing concentrations (0.1, 0.5 and 1 mmol/L) for 8 h, the MIP-1α mRNA expression was determined by in situ hybridization using a MIP-1α cDNA probe, and the MIP-1α protein expression was measured by cell enzyme-linked immunosorbent assay (ELISA) using a goat anti-human MIP-1α monoclonal antibody. RESULTS: The in situ hybridization showed that cultured HUVECs were able to express MIP-1α mRNA at a low level that was purplish blue granules in cytoplasm. After exposure to HCY at the concentrations mentioned above, the expression of MIP-1α mRNA was significantly increased in a dose-dependent manner. Analysis of variance showed that there was significant difference between groups ( F= 606.38, P< 0.01). Cell ELISA demonstrated that the MIP-1α protein expression in HUVECs of the experimental groups was also significantly enhanced by HCY in a dose-dependent manner. There was also significant difference between groups ( F =19.5, P< 0.01). CONCLUSION: These results demonstrate that HCY induces HUVECs to express MIP-1α mRNA and protein, which may play an important role in the recruitment of monocytes into arterial intima.更多还原