同型半胱氨酸诱导人脐静脉内皮细胞表达RANTES蛋白

为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RANTES蛋白 ,使人脐静脉内皮细胞暴露于不同浓度的同型半胱氨酸孵育 8h后 ,用免疫细胞化学和Westernblot方法检测RANTES蛋白的表达。免疫细胞化学检测结果发现 ,培养的人脐静脉内皮细胞能表达RANTES蛋白。培养的内皮细胞与 0 .1、0 .5及 1mmol L同型半胱氨酸共同孵育 8h后 ,其RANTES蛋白表达的平均吸光度值分别为 0 .0 4 34± 0 .0 0 6 3、0 .0 788± 0 .0 0 5 3和 0 .10 6 1± 0 .0 2 15 ,均显著高于对照组 (0 .0 2 0 0± 0 .0 0 32 ) ,方差分析发现 ,组间均存在显著性差异 (P <0 .0 1)。Westernblot检测结果发现 ,当内皮细胞与 0 .1、0 .5和 1mmol L同型半胱氨酸共同孵育 8h后 ,其免疫染色条带的积分吸光度值分别为 8873、10 2 0 0和 10 80 0 ,分别是对照组 (3881)的 2 .2 9倍、2 .6 3倍和 2 .78倍。此结果提示 ,培养的人脐静脉内皮细胞表达低水平的RANTES蛋白 ,同型半胱氨酸能提高其RANTES蛋白的表达

Homocysteine Induces the Expression of Regulated upon Activation, Normal Txpressed and Secred Protein in Human Umbilical Vein Endothelial Cells

Aim To investigate whether homocysteine (HCY) can induce cultured human umbilical vein endothelial cells (hUVEC) to express regulated upon activation, normal T expressed and secreted (RANTES) protein. Methods After exposure of the cultured hUVEC to HCY at increasing concentrations for 8 h, the RANTES protein expression was determined by immunocytochemistry and Western blot analysis. Results Cultured hUVEC could express RANTES protein. Immunocytochemistry showed the mean absorbance values of RANTES protein expression in hUVEC exposed to HCY at different concentrations (0.1, 0.5 and 1 mmol/L HCY) for 8 h were 0.0434±0.0063, 0.0788±0.0053 and 0.1061±0.0215, respectively, which were significantly higher than that of the control group (0.0200±0.0032). Analysis of variance proved a significant difference between groups (F=319.03, p<0.01). Western blot analysis displayed that exposure of hUVEC to HCY at gradient concentrations (0.1, 0.5 and 1 mmol/L) for 8 h resulted in a 2.29 fold, a 2.63 fold and a 2.78 fold increase in the expression of RANTES protein in the cells, compared with the control group. Conclusions The cultured hUVEC could express RANTES protein, and HCY was able to induce hUVEC to express RANTES protein at a higher level.