三七总皂苷对氧化型低密度脂蛋白诱导的人脐静脉内皮细胞血管细胞黏附分子1表达的影响*☆◆

背景：氧化低密度脂蛋白能诱导血管内皮细胞活化和黏附分子表达，在动脉粥样硬化形成早期起重要作用。三七总皂苷在心血管系统方面具有保护血管内皮细胞、显著改善动脉粥样硬化病变程度的药理作用。目的：验证三七总皂苷对氧化型低密度脂蛋白损伤内皮细胞后血管细胞黏附分子1的表达及与人单核细胞黏附的影响。设计、时间及地点：体外实验，分组对照，于2007-03/2008-05 在北京中医药大学东直门医院中医内科学教育部重点实验室完成。材料：原代人脐静脉内皮细胞为美国Cascade Biologics公司产品；三七总皂苷(血塞通冻干粉针)为黑龙江珍宝岛制药有限公司产品，成分为三七总皂苷，批号：20040207。方法：以培养原代人脐静脉内皮细胞作为靶细胞, 用氧化型低密度脂蛋白造成人脐静脉内皮细胞损伤模型。将培养的人脐静脉内皮细胞分为5组：氧化型低密度脂蛋白组(100 mg/L)、氧化型低密度脂蛋白 + 三七总皂苷组(终浓度分别为200，100， 50 mg/L)、正常对照组。主要观察指标：光镜下观察细胞形态变化, 四甲基偶氮唑盐法检测细胞活性；采用蛋白定量法检测人脐静脉内皮细胞与单核细胞的黏附率；用流式细胞仪测定人脐静脉内皮细胞的血管细胞黏附分子1的蛋白表达。结果：氧化型低密度脂蛋白作用人脐静脉内皮细胞后12，24 h时，人脐静脉内皮细胞损伤明显，细胞活性显著降低，人单核细胞与人脐静脉内皮细胞的黏附率显著升高, 人脐静脉内皮细胞的血管细胞黏附分子1的蛋白表达水平也均明显升高，均明显高于正常对照组(P < 0.05，P < 0.01)，而三七总皂苷能使氧化型低密度脂蛋白损伤的人脐静脉内皮细胞形态趋于正常，活性增强，并明显降低人单核细胞与人脐静脉内皮细胞的黏附率，以及显著降低血管细胞黏附分子1的蛋白的表达水平，与氧化型低密度脂蛋白组相比均有显著性差异(P < 0.05，P < 0.01)，这种作用随着剂量的增加而增强。结论：三七总皂苷能通过下调血管细胞黏附分子1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，这可能是其治疗动脉硬化闭塞症的机制之一。

Effects of panax notoginseng saponins on expression of vascular cell adhesion molecule-1 in human umbilical vein endothelial cells injury induced by oxidized low density lipoprotein

BACKGROUND: Oxidized low-density lipoprotein (ox-LDL) can induce the expression of vascular cell adhesion molecule-1 (VCAM-1), which plays an important role in the early stage of atherogenesis. Panax notoginseng saponins (PNS) protects human umbilical vein endothelial cells (HUVEC) against injury, improves the atherosclerotic lesions in cardiovascular system.OBJECTIVE: To verify the effect of PNS on the expression of VCAM-1 and the adherence to monocytes of cultured HUVEC injury induced by ox-LDL. DESIGN, TIME AND SETTING: The in vitro grouping and controlled experiment was performed at the key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing, Dongzhimen Hospital, Beijing University of Chinese Medicine between March 2007 and May 2008.MATERIALS: HUVEC was provided by American Cascade Biologics Company. PNS was supplied by Heilongjiang Zhenbaodao Pharmaceutical Co., Ltd (Lot number: 20040207).METHODS: Using the cultured HUVEC as target cells, the ox-LDL was used to establish injured model of HUVEC. The cells were divided into five groups. Except the control group, HUVEC cells were cultured with ox-LDL (100 mg/L), ox-LDL plus PNS with different concentrations (200, 100, 50 mg/L).MAIN OUTCOME MEASURES: The morphologic changes of HUVEC were observed under the light microscope and HUVEC activity was determined by MTT method. The adhesive percentage between HUVEC treated with ox-LDL and monocytes was determined by protein quantification. Expression of protein of VCAM-1 was determined by flow cytometry.RESULTS: Treatment of HUVEC with ox-LDL for 12, and 24 hours could significantly reduced the activity of HUVEC, increased HUVEC damages and adhesion of monocytes to HUVEC, and enhanced the protein expressions of VCAM-1, all the changes were higher than the normal control group (P < 0.05, P < 0.01). PNS could significantly increase the activity of HUVEC and lessened the injury of HUVEC, inhibited the adhesion between monocyte and endothelial cells, as well as the protein expressions of VCAM-1 in HUVEC. Compared with the ox-LDL group, there was significant difference (P < 0.05, P < 0.01). The effects were strengthened with increasing the deal of PNS.CONCLUSION: PNS has a protective effect on HUVEC via down-regulating the expression of VCAM-1, inhibiting the adhesion between monocyte and endothelial cells, which may be one of mechanisms why PNS can treat with arteriosclerosis obliterans.