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目的用葡萄籽原花青素(GSP)研究对H2O2诱导的脐静脉内皮细胞(HUVEC)损伤的保护作用及其机制。方法建立氧化损伤的细胞模型,体外培养HUVEC分为正常对照组、氧化损伤组以及高、中、低剂量(100,50,10μg/ml)GSP组;采用四甲基偶氮唑盐比色法(MTT法)检测GSP对H2O2诱导的内皮细胞氧化损伤的保护作用;逆转录聚合酶链反应(RT-PCR)检测单核细胞趋化蛋白(MCP-1)mRNA的水平;流式细胞术Annexin V/PI检测细胞凋亡以及定量检测各实验组细胞间粘附分子(ICAM-1)、血管细胞粘附分子(VCAM-1)的表达。结果内皮细胞氧化损伤后吸光度(OD)低于正常对照组;GSP预处理后OD值增加,高剂量GSP组的OD值与正常组相比无显著性差异;损伤组MCP-1 mRNA表达水平与内参照灰度值之比高于正常组;药物预处理氧化损伤后明显减弱。损伤组中细胞凋亡率显著增高、表达ICAM-1、VCAM-1的阳性细胞数均多于正常组;预处理后,凋亡细胞数以及两种粘附分子阳性细胞数明显减少,且呈剂量依赖性效应。结论葡萄籽原花青素可通过抑制内皮细胞粘附分子与细胞因子的表达而抑制炎症性损伤及其凋亡,达到保护内皮细胞的目的。
Objective To study the protective effect of grape seed proanthocyanidins (GSP) on H2O2-induced injury of human umbilical vein endothelial cells (HUVECs) and its mechanism. Methods HUVECs were divided into normal control group, oxidative injury group and high, medium and low dose GSP groups (100, 50, 10μg / ml) MTT assay was used to detect the protective effect of GSP on H2O2-induced endothelial cell oxidative damage. The level of MCP-1 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) V / PI was used to detect the apoptosis and quantitatively detect the expression of intercellular adhesion molecule (ICAM-1) and vascular cell adhesion molecule (VCAM-1) in each experimental group. Results The OD value of endothelial cells after oxidative injury was lower than that of the normal control group. The OD value of GSP pretreatment increased, while the OD value of high dose GSP group had no significant difference compared with normal group. The expression of MCP-1 mRNA in injury group was significantly correlated with The ratio of internal reference gray value was higher than that of normal group. The drug pretreatment significantly decreased after oxidative damage. The apoptosis rate of ICAM-1 and VCAM-1 positive cells in the injury group was significantly higher than that in the normal group. After pretreatment, the number of apoptotic cells and the number of positive cells of both adhesion molecules were significantly decreased Dose-dependent effects. Conclusion Grape seed proanthocyanidins can inhibit the inflammatory injury and apoptosis by inhibiting the expression of endothelial cell adhesion molecules and cytokines and achieve the purpose of protecting endothelial cells.