失血性休克大鼠体外实验关于内皮细胞蛋白C系统的研究

An in vitro study of thrombomodulin-protein C-EPCR system coagulation function in hemorrahgicshock rats

目的观察失血性休克大鼠血清调节内皮细胞表达凝血一抗凝分子TM和EPCR的作用。方法本实验于温州医学院实验室完成，选取成年健康SD大鼠30只随机（随机数字法）分成3组：健康对照组、实验对照组（假休克组）和实验休克组（失血性休克组），每组均为10只。采用颈内动脉均放血至40mmHg（1mmHg=0．133kPa）60min建立失血性休克动物模型，留取假休克组、休克组血清，并将胎牛血清作为健康对照组，无菌处理后加入到培养基，与内皮细胞分别培养6h、12h和18h，用逆转录-聚合酶链反应检测休克血清刺激后内皮细胞TM、EPCRmRNA的表达。结果与健康对照组相比，休克组血清刺激SVAREC6h后，TM和EPCRmRNA均升高（P〈0．01），18h升高仍具有统计学意义。结论失血性休克血清上调内皮细胞TM、，EPCRmRNA表达。血栓调节素-活化蛋白C-内皮细胞蛋白C受体系统发挥抗凝作用，保护内皮细胞功能。

Objective To investigate the coagulant function of Thrombomodulin-Protein C-EPCR System in rats with hemorrhagic shock in vitro study. Methods In this study, 30 rats were divided into three groups, with each group 10 rats： control group, the sham shock group, hemorrhagic shock group. Hemorrhagic shock model was subjected to computer-controlled arterial hemorrhage to 40 ram Hg for 60 min The rat blood serum samples were obtained from rats in sham shock group and shock group after maintaining 3 hours, and fetal bovine serum （FBS） was used as control group. Then medium contained fore-mentioned serums cultured with SV40-transformed aortic rat endothelial cells for 6 hours, 12 hours and 18hours. The messenger RNA （mRNA） expression of thrombmodulin （TM）, Endothelial cell protein C receptor in the SV40-transformed aortic rat endothelial cells which were treated with serums were detected by reverse-transcription polymerase chain reaction. Results Compared to control group, SVARECs were incubated for 6hrs with shock serum, the mRNA expression of TF, TM and t-PA was significantly higher, and continued elevating at the time of 18 hours （P 〈 0. 01 ）. Conclusions Serums from hemorrhagic shock rats upregulate TMmRNA EPCRmRNA expression on the endothelial cells surface. It is suggested that Thrombomodulin-Protein C-EPCR System pay a key role of anticoagulation, the protection of endothelial function.