脂质过氧化对培养的人内皮细胞血管细胞粘附分子-1表达的影响

Effect of lipid peroxidation on expression of vascular cell adhesion molecule-1 in cultured human endothelial cells

目的 :观察脂质过氧化对培养的人内皮细胞血管细胞粘附分子 - 1表达的影响。方法 :培养的人脐静脉内皮细胞随机分为实验组和对照组。实验组与不同浓度联胺 (1μmol/L、5 μmol/L、10 μmol/L)作用 8h ,对照组不加联胺。细胞原位杂交和细胞酶联免疫吸附法分别测定内皮细胞血管细胞粘附分子 - 1mRNA和蛋白表达。结果 :细胞原位杂交结果图像分析表明 ,实验组平均吸光度值分别为 0 .14 7± 0 .0 13、0 .2 92± 0 .0 2 0和 0 .396± 0 .0 2 2 ,是对照组 (0 .0 77± 0 .0 11)的 1.91倍、3.79倍和 5 .14倍。方差分析表明 ,各组间两两比较有显著差异 (P <0 .0 1)。细胞酶联免疫吸附测定结果 ,实验组平均吸光度值分别是 0 .15 8± 0 .0 0 8、0 .2 2 0± 0 .0 17和 0 .32 1± 0 .0 2 3,为对照组 (0 10 4±0 0 16 )的 1.5 3倍、2 .12倍和 3.0 9倍。各组间两两比较有显著差异 (P <0 .0 1)。结论 :人脐静脉内皮细胞脂质过氧化增加血管细胞粘附分子 - 1mRNA和蛋白表达 ,这可能促进单核细胞粘附血管内皮 ,在动脉粥样硬化发生发展中起重要作用。

AIM: To observe the expression of vascular cell adhesion molecule-1 (VCAM-1) in cultured human umbilical vein endothelial cells (HUVEC) by lipid peroxidation injury induced by exposure to diamide. METHODS: Expression of VCAM-1 mRNA and protein in HUVEC was determined by in situ hybridization and a cell enzyme-linked immunosorbent essay (cell ELISA), respectively. RESULTS: The HPIAS-1000 image analytic system in situ hybridization detected that the mean absorbance values in experiment groups(1, 5 and 10 μmol/L diamide for 8 hours) were 0.147±0.013, 0.292±0.020 and 0.396±0.022, which were 1.91-fold, 3.79-fold and 5.14-fold as much as that of the control group (0.077±0.011), respectively. There was significant statistical difference between groups ( P< 0.01). The cell ELISA showed that the mean absorbance values of VCAM-1 proteins in experiment groups were 0.158±0.008,0.220±0.017 and 0.321±0.023, which were 1.53-fold, 2.12-fold and 3.09-fold as much as that of the control group (0.104±0.016), respectively. The analysis of variance proved the significant difference between groups ( P <0.01). CONCLUSION: The results suggest that the increased expression of VCAM-1 is integral in promoting adhesion of monocytes to the vascular endothelium.