活血注射液对ox-LDL诱导的单核-内皮细胞黏附作用的影响

Effects of Huoxue Injection on adherence between human monocytes and vascular endothelial cells induced by oxidized low-density lipoprotein

目的观察活血注射液对氧化低密度脂蛋白(ox-LDL)诱导的人脐静脉内皮细胞(HUVEC)中细胞间黏附分子-1(ICAM-1)的表达及与人单核细胞黏附作用的影响。方法以培养HUVEC作为靶细胞,在内皮细胞培养基中加入ox-LDL制备细胞损伤模型。采用蛋白定量法检测HUVEC与单核细胞的黏附率;RT-PCR检测HUVEC中ICAM-1的mRNA表达;用流式细胞仪测定HUVEC中ICAM-1的蛋白表达。结果ox-LDL作用HUVEC后12、24h时,人单核细胞与HUVEC的黏附率显著升高,HUVEC中ICAM-1的mRNA和蛋白表达水平也均明显升高,均明显高于正常对照组(P<0.01),而活血注射液可明显降低人单核细胞与HUVEC的黏附率,以及显著降低ICAM-1的mRNA和蛋白的表达水平(P<0.05,P<0.01),这种作用随着剂量的增加而增强。结论活血注射液能通过下调内皮细胞表面黏附分子的表达抑制单核-血管内皮细胞黏附,从而发挥对血管内皮细胞的保护作用,有利于减少或抑制动脉粥样硬化的形成。

Objective To observe the effects of Huoxue Injection on the expression of intercellular cell adhesion molecule - 1 （ICAM-1） and the adherence to monocytes of cultured human umbilical vein endothelial cells （HUVEC） induced by oxidized low-density lipoprotein （ox-LDL）. Methods Cultured HUVEC was used as target cells; ox-LDL was added to the cell medium to prepare the injury model of human endothelial cell. The adhesive percentage of monocytes to HUVEC was detected by protein quantitative method. The expression of mRNA and protein of ICAM-1 were detected by RT-PCR and flow cytometry respectively. Results The adhesive percentage and the expressions of mRNA and protein of ICAM-1 were enhanced by ox-LDL after pretreating HUVEC for 12 h and 24 h（p 〈0.01, compared with the normal grounp） ; Huoxue Injection could significantly inhibit the adhesion between monocytes and HUVECs and the expression of mRNA and protein of ICAM-1 in HUVECs in a dose-dependent manner. Conclusion Huoxue Injection has an inhibitory effect on the adherence of monocytes to endothelial cells, probably by way of down - regulating the expression of mRNA and protein of ICAM-1 in endothelial cells, which can bring about the protective effect on endothelial cells injury induced by ox-LDL and decelerate the progression of atherosclerosis.