皖南蝮蛇毒PCA在DIC诊断中的应用

Application of the venom PCA extracted from Agkistrodon halys to diagnosis of DIC

目的:探讨皖南蝮蛇毒PCA在DIC诊断中的应用研究。方法:用脂多糖(LPS)100μg/(kg.h)(10 ml/h)的速度持续滴注6 h,建立稳定的兔DIC模型。分别于1 h、2 h、3 h、4 h、6 h时从股动脉取血1 ml,半自动凝血分析仪检测凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)和纤维蛋白原含量(FIB),发色底物法测定蛋白C(PC)的活性。观察DIC时PC活性水平变化。24 h内死亡的兔,立即取其肝、肾、肺标本;24 h观察期结束后,处死并取兔肝、肾、肺标本,均用10%甲醛固定,经包埋、切片后,行HE染色。结果:兔经持续耳缘静脉滴注LPS后,PC活性显著降低(LPS静脉滴注1 h时即已开始),PT与APTT明显延长、FIB明显降低(LPS静脉滴注2 h时开始)。LPS实验组兔肝、肾、肺病理切片显示细胞水肿、尚伴有灶性出血,肺微血管与肾小球亦见纤维蛋白微血栓形成。结论:本院蛇毒研究所研发的PC活性检测试剂有望成为评价DIC发展及诊断的检测试剂。

Objective ： To assess the application of protein C activator （PCA） extracted from the venom of Agkistrodon halys living in south Anhui province to diagnosis of disseminated intravascular coagulation （DIC）. Methods ： DIC models of rabbits were induced by continuous infusion of lipopolysaceharide （ LPS ） at velocity and dosage of 100 ug/（ kg . h） for 6 h. After animal preparation, 1 ml blood was taken via the femoral artery at 1 h, 2 h, 3 h, 4 h and 6 h, respectively. Semiautomatic coagulation analyzer was used to determine the prothrombin time（PT）, activated partial thromboplastin time（APTT） and fibrinogen content （ FIB ）, and ehromogenic substrate assay was performed to measure the activity of PCA and its level change in DIC. For the experimental rabbits died within 24 h, the samples of liver, lung and kidney were taken and fixed in 4% paraformaldehyde, embedded in paraffin, serially sec- tioned and stained with hematoxylin eosin,and those survived 24 h by the end of experimental observation were also killed for the corresponding samples that were managed with the previous methods. Results ：By continuous infusion of LPS, PCA activity became degenerated, whereas PT and APTT were significantly extended. Clear decrease of FIB occurred at 2 h of LPS induction. The tissue sections from animals intervened with LPS revealed cellular edema in the liver, kidney and lung, with focal hemorrhage and microvascular fibrin micro-thrombosis in the lung and kidney glomerulus. Conclusion ： The reagent developed from the snake venom in our laboratory unit might serve as a kit to detection of the PC activity in estimation of DIC.