Corilagin抗PAF诱导血管内皮细胞损伤的保护作用

目的研究Corilagin抗血小板活化因子（PAF）损伤人脐静脉血管内皮细胞（HUVEC）的保护作用及其机制．方法用PAF损伤HUVEC，采用MTT法测定Corilagin对HUVEC活性的影响；以Fura-2／AM荧光探针负载细胞，用双波长荧光分光光度法测定HUVEC细胞内游离钙离子浓度；用ELISA法测定培养的细胞上清液中t-PA、PAI-1含量的变化．结果1μmol／L的PAF刺激内皮细胞2h后，可明显降低HUVEC的吸光度值，细胞内钙离子浓度升高，PAI-1含量明显升高，t-PA含量无明显变化；50～200μmol／L的Corilagin可升高PAF引起HUVEC吸光度值的降低，呈浓度依赖性显著降低HUVEC内游离钙浓度；Corilagin明显抑制PAF诱导的PAI-1分泌增高，对t-PA水平无明显影响．结论Corilagin对PAF诱导的HUVEC损伤具有保护作用，其机理可能与降低细胞内钙离子浓度、抑制PAI-1产生、调节t-PA／PAI-1平衡密切相关．

Protective Effects of Corilagin on Human Umbilical Vein Endothelial CeU Injury Induced by PAF

Objective To investigate the protective effect of corilagin on human umbilical vein endothelial cell （HUVEC） injury and its mechanism. Methods Platelet activating factor （PAF） was used to injure HUVEC in vitro. Cell viability was assessed using methyl thiazolyt tetrazolium （MTF） assay. The cultured cells were loaded by Fura-2/AM and the change of Ca2＋]i in HUVEC was measured by fluorospectrophotometry. The levels of tissue plasminogen activator （t-PA） and plasminogen activator inhibitor-1 （PAI-1） in the supernatant were measured hy enzyme linked immunosorbent assay （ELISA） , respectively. Results PAF significantly decreased the absorbance values of HUVEC, markedly elevated cytosolic calcium concentrations. PAI-1 level, as compared with the control group, was significantly increased by PAF. The concentrations of t-PA, however, had no significantly change in the supernatant of cultivated cells. Corilagin significantly inhibited PAF-indueed Ca2＋]i elevation in a concentration-dependent manner. Corilagin （50 -200 μmol/L） significantly reduced the level of PAI-1, while had no effect on t-PA level. Gonclusion Corilagin can protect against the injury induced by PAF in cultured HUVEC. The mechanism may be associated with suppressing the mobilization of cytosolic calcium, inhibiting the generation of PAI-1, and modulating the balance of tPA/PAI-1.