绞股蓝总苷对人脐内皮细胞损伤的保护作用

Protective effect of Gypenosides on human vascular endothelial cells injury induced by cholesterol in vitro

目的研究绞股蓝总苷(GPs)对胆固醇所致人脐静脉内皮细胞(hVECs)损伤的保护作用及机制。方法以体外培养的hVECs-12为研究对象,设正常对照组、溶媒组(二甲基亚砜)、胆固醇组(以50mg/L胆固醇处理48h)、3种浓度绞股蓝总苷(1、10、100μg/ml)组(按剂量在培养基中加入绞股蓝总苷预处理1h后再加入50mg/L的胆固醇处理48h)。分别用比色法、硝酸酶还原法及ELISA法检测细胞培液中乳酸脱氢酶(LDH)活性、一氧化氮(NO)浓度、单核细胞趋化蛋白-1(MCP-1)浓度;以DCFH-DA为荧光探针,流式细胞术检测细胞内活性氧水平;免疫细胞化学染色方法检测细胞内核因子-κBp65(NF-κBp65)核移位阳性细胞数。结果与正常对照组比较,胆固醇组各指标差异均有统计学意义(P<0.01);与胆固醇组比较,GPs可减少细胞培液中LDH活性、MCP-1浓度、并能增加NO浓度(P<0.01)、可降低细胞内ROS的水平(P<0.01);GPs能抑制NF-κB核移位(P<0.01),且随浓度升高有递减趋势。结论绞股蓝总苷对胆固醇所致的人脐静脉内皮细胞损伤具有保护作用,机制可能与减少细胞内ROS的生成,从而抑制NF-κB活化有关。

Objective To investigate the protective effect of Gypenosides on the injury of human vascular endothelial ceels（hVECs）induced by cholesterol and its mechanisms.Methods hVECs-12 were cultured in vitro and treated with or without cholesterol,and with different concentrations（1,10,100 μg/ml）GPs before treating with cholesterol for 48 hours.LDH activity and concentration of nitric oxide in the supernatant of cell culture medium were detected relatively by colorimetry;the concentration of MCP-1 protein in cell culture medium was detected by ELISA;nucleic translocation of NF-κBp65 was detected by immunocytochemistry staining.At the same time,intracellular ROS level was determined by flow cytometry（FCM）with DCFH-DA as fluorescent probe.Results Compared with the cholesterol group,GPs could significantly decrease LDH activity and the MCP-l protein level,and markedly increase the NO level.In addition,GPs could significantly decrease the level of intracellular ROS（P0.01）;GPs could restrain the nuclear translocation of NF-κB in hVECs.Conclusions GPs can protect hVECs from injury due to cholesterol by decreasing ROS level in endothelial cells and inactivation of NF-κB.