蛇伤胶囊对竹叶青蛇伤血管内皮功能影响

Influence of Sheshang Capsule on Vascular Endothelial Function in Trimeresurus Stejnegeri Snakebites

目的:研究蛇伤胶囊治疗竹叶青蛇伤凝血障碍的机制。方法:根据前期实验复制竹叶青蛇伤的动物及细胞模型。均设置空白组(KB)、模型组(MX)和低、中、高剂量蛇伤胶囊药液组(DY、ZY、GY)5组,每组10个样本。在动物实验中,KB经兔右后腿皮下注射0.75 m L/kg生理盐水,6 h后灌胃10 m L/kg生理盐水。MX、DY、ZY和GY经兔右后腿皮下注射0.75 m L/kg竹叶青蛇毒液,6 h后分别灌胃10 m L/kg生理盐水和10 m L/kg低、中、高剂量蛇伤胶囊药液。各组均每日灌胃1次,连续灌胃1周。于末次灌胃后24 h经耳缘静脉采血,分离出血清。在细胞实验中,KB加入10%正常兔血清培养,MX在KB的基础上加入5μg/m L竹叶青蛇毒液培养,DY、ZY和GY在MX基础上培养6 h后分别加入5%、10%和15%的含中药兔血清继续培养。各组分别培养72 h后,收集细胞培养液及细胞。以酶联免疫吸附法检测兔血清及细胞培养液中的内皮素-1(Endothelin-1,ET-1)、一氧化氮(Nitric Oxide,NO)和血管性血友病因子(von Willebrand Factor,v WF)水平。流式细胞术检测细胞中NO含量。结果:MX的ET-1和v WF水平相对KB均出现了显著的升高(P<0.01,P<0.05),NO水平显著降低(P<0.01);DY、ZY和GY的ET-1及v WF水平相对MX均出现不同程度降低,NO水平出现不同程度的升高,其中ZY组变化有统计学意义(P<0.01或P<0.05)。结论:蛇伤胶囊可通过抗血管内皮细胞损伤途径治疗竹叶青蛇伤凝血障碍。

Objective：To investigate the mechanism of Sheshang Capsule treating the coagulation disorder induced by Trimeresurus stejnegeri snakebites. Methods：The animal and cell models were established according to the former study. The blank group（KB）,the model group（MX）and the low,middle and high dose Sheshang Capsule liquid group（DY,ZY,GY）were set up. There were 10 samples in each group. In vivo,KB was injected 0.75 m L/kg normal saline into rabbits＇ right hind legs,and received 10 m L/kg normal saline by gavage after 6 hours. MX,DY,ZY and GY were injected 0.75 m L/kg Trimeresurus stejnegeri venom into rabbits＇ right hind legs. After 6 hours,MX received 10 m L/kg normal saline by gavage,and Sheshang Capsule groups with different doses received 10 m L/kg low,intermediate and high dose Sheshang Capsule liquid by gavage respectively. The gavage was given once daily for a week. 24 hours after the last gavage,the blood of the rabbits was collected through auricular vein and the serum was separated. In vitro,KB was cultured with 10% blank rabbit serum. MX was cultured with 5 μg/m L of snake venom in addition. DY,ZY and GY were cultured with the same composition as MX in the first 6 hours and then they were cultured with 5%,10% and 15% drug serum respectively in addition. After 72 hours culture,the medium and cells were collected. The level of endothelin-1（ET-1）,nitric oxide（NO）andvon Willebrand factor（v WF）in the serum and medium were measured by enzyme linked immunosorbent assay. The contents of NO in cells were measured by flow cytometry. Results：The content of ET-1 and vW F increased and NO decreased in MX significantly compared with KB（P〈0.01 or P〈0.05）. In DY,ZY and GY,the level of ET-1 and vW F all decreased and NO all increased compared with MX. The variations in ZY were remarkble（P〈0.01 or P〈0.05）. Conclusion：Sheshang Capsule could treat the coagulation disorders induced by Trimeresurus stejnegeri snakebites. It is realized by the way of anti vascular endothelial cell injury.