联胺对培养的人脐静脉内皮细胞表达细胞间粘附分子-1的影响

The Influence of Diamide on Expression of Intercellular Adhesion Molecule-1 in Cultured human Umbilical Vein Endothelial Cells

人脐静脉内皮细胞 (HUVEC)暴露于 1、 5和 10 μmol/L 联胺作用 8h,或 5 μmol/L 联胺作用 4和 2 4 h,观察其细胞间粘附分子 - 1(ICAM- 1)蛋白表达。免疫细胞化学法 :实验组平均吸光度值分别是对照组的 1.4 9、 1.80、 2 .2 4、1.5 3和 2 .4 4倍 (F=16 0 .2 7,P<0 .0 1)。流式细胞仪分析 :实验组荧光抗体阳性细胞百分率分别是 32 .9%、 4 3.0 %、5 0 .9%、 36 .0 %和 5 2 .4 % ,高于对照组 (2 1.4 % )。不同浓度组间进行 χ2检验 ,有统计学意义 (χ2 =2 0 98.2 3,P<0 .0 0 1;R=- 0 .2 2 8,P<0 .0 0 1) ,或不同时间组间 (χ2 =2 16 7.76 ,P<0 .0 0 1;R=- 0 .192 ,P<0 .0 0 1)。 Western Blot检测 ,实验组积分吸光度值分别是对照组的 1.2 3、 1.75、 1.92、 1.5 6和 1.82倍。结果提示 :联胺诱导 HUVEC表达

Oxidant diamide induced expression of intercellularadhesion molecule 1 (ICAM 1) in human umbilical vein endothelial cells (HUVECs) was observed. After exposure of the hUVECs to diamide at 1, 5 and 10 μmol/L for 8 h, or to 5 μmol/L for 4 and 24 h, the expression of ICAM 1 protein was detected by immunocytochemistry assay, flow cytometric technique and Western blotting. Immunocytochemistry assay showed the mean absorbance values in experimental groups were 1 49 , 1 80 , 2 24 , 1 53 and 2 44 fold as much as that in the control group ( F=160 27, P <0 01). Flow cytometry revealed that the percentages of ICAM 1 fluorescence labeling cells in experimental groups were 32 9, 43 0, 50 9, 36 0 and 52 4, respectively, which were higher than in the control group (21 4). The χ 2 test of variance proved the significant difference among 1 μmol/L, 5 μmol/L, 10 μmol/L and the control group (χ 2=2098 23, P<0 001;R=-0 228,P <0 001) or among 4 h, 24 h and the control group (χ 2=2167 76, P<0 001;R=-0 192,P <0 001). Western blotting showed that the integral absorbance values of ICAM 1 protein in experimental groups were 1 23 , 1 75 , 1 92, 1 56 and 1 82 fold as much as that in control group, respectively. These results indicated that after exposure of the hUVECs to diamide the ICAM 1 protein expression was increased in a dose and time dependent manner. The upregulated ICAM 1 expression may promote the adherence of blood monocyte to the endothelium and play an important role in the recruitment of monocytes into the intima and atherogenesis.