滋阴活血解毒方对AGEs诱导VEC损伤黏附分子及凝血相关因子表达的影响

摘要

目的 观察滋阴活血解毒方对糖基化终末产物(advanced glycation end-producs,AGEs)诱导血管内皮细胞(vas-cular endothelial cell,VEC)损伤的影响及其黏附分子、凝血相关因子表达的变化,进一步探讨该方对AGEs诱导VEC损伤的保护作用.方法 以人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)作为实验对象,以终浓度(100 mg/L)的AGEs诱导HUVEC损伤.各模型组及给药组细胞分别继续培养12、24、48 h.在显微镜下观察各组细胞形态变化,并以Western-blot法检测各组黏附分子VCAM-1、ICAM-1的表达;以RT-PCR方法 检测TF、TM mRNA表达.结果 相对于空白组细胞,模型组细胞出现异常增殖,细胞数量明显增多,多数细胞变形皱缩,胞膜轮廓模糊,细胞内出现粗糙样颗粒变化,VCAM-1、ICAM-1、TF表达明显上调,TM表达下调,且在作用24 h后差异有显著性意义(P<0.01);相同时间点,给药组较模型组圆缩形细胞减少,能下调VCAM-1、ICAM-1、TF表达,上调TM表达,且随着作用时间的延长更明显(P<0.01).结论 滋阴活血解毒方能抑制AGEs对VEC的损伤,降低黏附分子的表达,下调表达组织因子mRNA转录,上调血栓调节素mRNA转录.%Objective To investigate the influence of Ziyin Huoxue Jiedu decoction on the expression of adhe-sion molecules and blood coagulation related factors in the vascular endothelial cells (VEC) induced by advanced glycation end-products (AGEs). Methods The human umbilical vein endothelial cell as the subjects, the human umbilical vein endothe-lial cells (HUVEC) damage were induced by AGEs with the final concentration (100 mg/L), and the cells were continually cultured for 12, 24, 48 h. The cell morphological changes were observed, the dhesion molecule VCAM-1 and ICAM-1 ex-pression were determined by western-blot. The TF and TM mRNA expression were detected by RT-PCR method. Results Compared with the blank group, the model group cell was abnormal proliferation, cell number increased obviously, most of the cell deformation shrinkage, cell membrane hazy outline, a rough sample are observed in the cell. The VCAM 1, ICAM-1, TF expression significantly raised, TM expression reduced. There was significant diffrence after 24 h (P<0.01). Compared with the model group, the VCAM-1, ICAM-1, TF expression in medicine group down-regulated, TM expression up-regulated, th e effect was stronger along with time(P<0.01). Conclusion Ziyin Huoxue Jiedu decocotion could inhibit the damage of VEC in-duced by AGEs, reduce the expression of adhesion molecules, down-regulate the expression of tissue factor mRNA transcrip-tion, up-regulate thrombomodulin mRNA transcription.