大鼠主动脉内皮损伤后核因子κB活性的动态变化及血管细胞黏附分子1的表达

Time Course of Nuclear Factor-κB activation and Vascular Cell Adhesion Molcular-1 Expression in Rat Artery After Endothelium Denudation

目的观察动脉损伤后核因子κB活性的动态变化,以及血管局部血管细胞黏附分子1的表达,分析二者的关系。方法SD大鼠以球囊剥脱主动脉内皮建立大鼠动脉损伤模型。按照术后处死的时间点,将动物分为8组：0 h（对照组）、12 h、24 h、1天、2天、3天、7天及14天,每组6只。凝胶迁移率实验检测各个时间点核因子κB活性的动态变化,免疫组织化学观察血管细胞黏附分子1在血管局部的表达。结果术后即刻以扫描电镜观察内皮完全剥脱。术后14天可见明显的增生内膜,模型建立成功。大鼠主动脉球囊损伤后,核因子κB在术后12 h即有明显活化,高峰在13天,然后逐渐下降,14天左右接近术前水平。术后即刻无血管细胞黏附分子1表达,7天时在内膜表面及少量中膜平滑肌细胞可以看到血管细胞黏附分子1表达,术后14天时在未有内皮细胞覆盖处,新生内膜的近血管腔面少量表达,而主要的阳性转移到新生内膜近内弹力板处。结论核因子κB和血管细胞黏附分子1参与大鼠的主动脉损伤后的炎症过程;核因子κB的活化与失活成动态变化;血管细胞黏附分子1不同时间表达在不同的部位,核因子κB的活化与失活在血管细胞黏附分子1的表达与消退之前,核因子κB对血管细胞黏附分子1的表达起调节作用。

Aim To investigate time course of nuclear factor-κB（NF-κB） activation after artery endothelium denudation and its relationship with local vascular cell adhesion molcular-1（VCAM-1） expression. Methods Deendothelialization injury of rat aorta was produced by ballooning（3 times） with PTCA catheter.Crucify 6 rats before and at every time points after procedure： 0 h,12 h,24 h,1 d,2 d,3 d,7 d and 14 d.NF-κB activation was measured by electrophoretic mobility shift assay（EMSA）.Local VCAM-1 expression was measured by immunohistochemical Assay. Results Scanning electron microscopy showed that endothelium was completely denudated.Significant neointima proliferation was seen at 14 days after injury.NF-κB was activated at 12 h and culminated at 1 to 3 days after operation,and then decreased gradually.Its activation was higher at 14 days after operation than control,but there was no statistical significance.No local VCAM-1 expression could be detected immediately after denudation.At 7 days after operation,VCAM-1 was positive on the surface of endothelium and in the medium smooth muscle cells（SMC）.At 14 days after operation,VCAM-1 was still positive on the surface of uncovered endothelium,while more abundant expression was seen in the bottom of neointima near internal elastic lamina. Conclusions NF-κB activation and VCAM-1 expression participate in the inflammation of rat denudated artery.Activation and inactivation of NF-κB occur before the expression and disappearance of VCAM-1.