摘要
探讨同型半胱氨酸是否可诱导人脐静脉内皮细胞表达巨噬细胞炎性蛋白 1α ,使培养的人脐静脉内皮细胞暴露于不同浓度的同型半胱氨酸 ,采用逆转录聚合酶链反应和免疫细胞化学方法观察其巨噬细胞炎性蛋白 1α的表达。结果发现 ,培养的人脐静脉内皮细胞可表达巨噬细胞炎性蛋白 1α。暴露于同一浓度 (0 .1mmol L)同型半胱氨酸 ,分别孵育 4、8和 16h ,其巨噬细胞炎性蛋白 1αmRNA表达水平分别是对照组的 2 .0 8倍、3.2 6倍和 3.35倍 ;免疫细胞化学发现 ,上述各组巨噬细胞炎性蛋白 1α蛋白表达的平均吸光度值分别是 0 .0 71± 0 .0 0 6、0 .0 81± 0 .0 0 6和 0 .12 8± 0 .0 0 9,均显著高于对照组 (0 .0 4 9± 0 .0 0 5 ,P <0 .0 1)。暴露于不同浓度 (0 .1mmol L、0 .5mmol L和 1mmol L)的同型半胱氨酸 ,孵育 8h后 ,各组巨噬细胞炎性蛋白 1αmRNA表达水平分别是对照组的 2 .0 8倍、4 .35倍和 4 .5 7倍 ,巨噬细胞炎性蛋白 1α蛋白表达的平均吸光度值分别是 0 .0 81± 0 .0 0 6、0 .110± 0 .0 0 9和 0 .118± 0 .0 0 8,均显著高于对照组 (0 .0 4 9± 0 .0 0 5 ,P <0 .0 1)。结果提示 ,同型半胱氨酸可诱导人脐静脉内皮细胞表达高水平的巨噬细胞炎性蛋白 1αmRNA和蛋白。
Aim To investigate whether homocysteine (HCY) could induce human umbilical vein endothelial cells (hUVEC) to express macrophage inflammatory protein 1α (MIP 1α). Methods After exposure of the cultured hUVEC to HCY either at a same concentration for different incubation time or at increasing concentrations but for a same incubation time, the MIP 1α mRNA and protein expression in the cells were determined by reverse transcriptase polymerase chain reaction (RT PCR) and immunocytochemistry, respectively. Results Cultured hUVEC could express MIP 1α mRNA and protein. RT PCR showed that exposure of hUVEC to HCY at a concentration of 0.1 mmol/L for 4, 8 and 16 h, resulted in a 2.08 fold, a 3.26 fold and a 3.35 fold increase in the expression of MIP 1α mRNA in hUVEC, compared with the control group; meanwhile, exposure of hUVEC to HCY at different concentrations (0.1 mmol/L, 0.5 mmol/L and 1 mmol/L) for 8 h, resulted in a 2.08 fold, a 4.35 fold and a 4.57 fold increase in the expression of MIP 1α mRNA in hUVEC as much as the control group, respectively. Immunocytochemistry showed that the absorbance values of MIP 1α protein expression in hUVEC exposed to HCY at a same concentration but for different incubation time mentioned above were 0.071±0.006, 0.081±0.006 and 0.128±0.009 (control group 0.049±0.005), respectively. There was a significantly statistical difference between groups (F=946.70, P<0.01). The absorbance values of MIP 1α protein in hUVEC exposed to HCY at different concentrations but for a same incubation time mentioned above were 0.081±0.006, 0.110±0.009 and 0.118±0.008 (control group 0.049±0.005), respectively, which also showeol significantly statistical difference between groups (F=546.89, P<0.01). Conclusion HCY was able to induce a high expression of MIP 1α mRNA and protein in cultured hUVEC.
出处
《中国动脉硬化杂志》
CAS
CSCD
2002年第2期101-104,共4页
Chinese Journal of Arteriosclerosis
基金
国家自然科学基金 (3 973 0 2 2 0 )
