摘要
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B：（TXB：）和6一酮一前列腺素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、600mg／L的水蛭提取液加入到10kU／L凝血酶刺激的细胞中培养12h，空白对照组加入等量RPMI1640培养液。取上清液，采用酶联免疫吸附试验（ELISA）检测TXB：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—PGFl。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—PGF1α58．67±3．24比21．31±1．12，均P〈O．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。
Objective To investigate the anti-coagulation mechanism of Hirudo extract on the release of thromboxane B2 （TXB2） and 6-keto-prostaglandin FI = （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- PGFI, 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-PGF1, level （ng/L） was significantly lower （21.31 _＋ I. 12 vs. 30.54 ＋__ 1. I I, 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-PGF~, （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-PGF~, ： 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-PGF1 a, 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.
出处
《中国中西医结合急救杂志》
CAS
北大核心
2013年第3期146-148,共3页
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care
基金
湖南省自然科学基金资助项目（11JJ5065）
湖南省大学生研究性学习和创新性实验计划资助项目（湘教通[2011]272号）
湖南中医药大学研究生科研创新资助项目（2012ex07）
湖南中医药大学“十二五”校级重点学科药物分析学资助项目
