水蛭提取液对凝血酶诱导血管内皮细胞释放血栓素B2和6-酮-前列腺素F1α的影响

摘要

Objective To investigate the anti-coagulation mechanism of Hirudo extract on the release of thromboxane B2(TXB2)and 6-keto-prostaglandin F1α(6-keto-PGF1α)induced by thrombin from cultured human umbilical vein endotheliai cells(HUVECs). Methods The HUVECs were cultured in vitro. The well grown HUVECs of 2-3 generation were categorized into different kinds of groups:blank control group,thrombin-simulated group and groups of Hirudo extracts with low,medium and high doses. The raw drug concentrations of 150,300,and 600 mg/L of Hirudo extract were separately added to thrombin-simulated cells,and then the mixture was cultured for 12 hours. At the same time,an equal amount of RMPI 1640 culture medium was added to the blank control group. The enzyme-linked immunosorbent assay(ELISA)was employed to measure the concentrations of TXB2 and 6-keto-PGF1α in the supernatant. Results Compared with the blank control group,TXB2 level(ng/L)in the stimulation group was significantly higher(206.53±18.60 vs. 115.21±12.31,P<0.01),and 6-keto-PGF1α level(ng/L)was significantly lower(21.31±1.12 vs. 30.54±1.11,P<0.01). Compared to the thrombin stimulated group,the TXB2 (ng/L)concentration in each of the Hirudo extract groups was markedly lowered and the 6-keto-PGF1α(ng/L)level was substantially increased,with the most significant changes found in the highest dose group(TXB2:109.18±9.72 vs. 206.53±18.60;6-keto-PGF1α:58.67±3.24 vs. 21.31±1.12,both P<0.01). Conclusions The Hirudo extracts could significantly weaken the inhibition effect of thrombin on HUVEC release of 6-keto-PGF1α, and could also resist the HUVEC release of TXB2 induced by thrombin . Its mechanism may be related to the regulation of balance between thromboxane and prostaglandin in the cells.%　　目的探讨水蛭提取液对凝血酶诱导血管内皮细胞（VEC）释放血栓素B2（TXB2）和6-酮-前列腺素F1α（6-keto-PGF1α）的作用机制。方法体外培养人脐静脉内皮细胞（HUVEC），将2～3代生长良好的细胞分为空白对照组、凝血酶刺激组和水蛭提取液低、中、高剂量组。将生药浓度分别为150、300、600 mg/L的水蛭提取液加入到10 kU/L凝血酶刺激的细胞中培养12 h，空白对照组加入等量RPMI 1640培养液。取上清液，采用酶联免疫吸附试验（ELISA）检测TXB2和6-keto-PGF1α的含量。结果与空白对照组比较，凝血酶刺激组TXB2含量（ng/L）明显升高（206.53±18.60比115.21±12.31，P＜0.01），6-keto-PGF1α含量（ng/L）明显降低（21.31±1.12比30.54±1.11，P＜0.01）；与凝血酶刺激组比较，水蛭提取液各剂量组TXB2（ng/L）含量均明显降低，6-keto-PGF1α（ng/L）含量均明显升高，且以高剂量组变化更显著（TXB2：109.18±9.72比206.53±18.60；6-keto-PGF1α：58.67±3.24比21.31±1.12，均P＜0.01）。结论水蛭提取液能明显减弱凝血酶诱导HUVEC释放6-keto-PGF1α的抑制作用，并能对抗凝血酶诱导HUVEC释放TXB2，其作用机制可能与其调节血栓素/前列环素平衡有关。