芹菜籽提取物保护Toll样受体4参与的内皮细胞过氧化损伤

目的观察芹菜籽提取物(CSE)在保护氧化性低密度脂蛋白(ox-LDL)诱导人脐静脉内皮细胞(HUVEC)损伤时,细胞生长与炎症反应Toll样受体4(TLR4)表达的影响,探讨CSE抗氧化损伤的可能作用机制。方法体外培养HUVEC,CSE三个浓度预处理,加入ox-LDL氧化损伤,应用四甲基偶氮唑盐比色法(MTT)测定细胞活力、流式细胞术PI染色检测细胞周期、激光共聚焦显微镜JC-1染色检测线粒体膜电位,Western blot检测TLR4蛋白水平的表达。结果与正常对照组相比,经ox-LDL损伤后HUVEC细胞吸光度(OD值)下降(P<0.05,P<0.01),且呈浓度依赖性效应、细胞周期阻滞在G0期,S期细胞比例下降(P<0.01)、细胞线粒体膜电位明显下降、TLR4的表达与正常组相比明显升高;CSE预处理使细胞吸光度上升、细胞周期S期细胞比例显著增加(P<0.05或P<0.01)、线粒体膜电位上升、TLR4的表达与损伤组相比下降(P<0.05或P<0.01)。结论抑制TLR4的表达可能是CSE抗内皮细胞氧化损伤、阻止经线粒体途径的细胞周期停滞的可能机制之一。

PROTECTION OF CELERY SEED EXTRACT ON TOLL LIKE RECEPTOR 4 INVOLVED OXIDATIVE DAMAGES OF ENDOTHELIAL CELLS

Objective To observe the protective effect of celery seed extract(CSE) on human endothelial cells(HUVEC) damaged by oxidized low density lipoprotein(ox-LDL) and discuss the anti-oxidative damage mechanism.Method HUVEC were cultured with CSE treatment in vitro.Methylthiazolyl tetrazolium(MTT) assay was used to detect cell vitality.Flow cytometry was used to detect cell cycle.Mitochondria membrane potential was detectal by laser scanning confocal microscope after JC-1 dyeing and Toll like receptor 4(TLR4) expression was tested by Western blot.Results Compared with the normal control group,cellular viability in the ox-LDL injuried group was decreased.The cell cycle was arrested in G0/G1 phase and the cell number of G0/G1 phase was increased(P<0.01),but the cell number in S phase was decreased(P<0.01).The mitochondrial membrane potential(ΔΨm) was obviously decreased.TLR4 expression was increased significantly.When pretreated with CSE,compared to injured group,the cellular viability was increased dose-dependantly(P<0.05 and P<0.01,respectively).The ratio of cell number in S phase was increased(P<0.05 and P<0.01,respectively) and mitochondria membrane potential also rose,TLR4 expression was significantly decreased(P<0.05 and P<0.01,respectively).Conclusion CSE can inhibit TLR4 expression of HUVEC,which is associated possibly with decresed mitochondrial oxidative damage and cell.