4种高毒有机磷化合物体外稳定性及小鼠染毒后体内活性持续时间  

Stability of four highly toxic organophosphorus compounds in vitro and their duration of activity in vivo after mouse exposure

作　　者：王淋 章子男 邢欢纯 曹文缤 张怡[3] 王永安 杨军 WANG Lin;ZHANG Zi-nan;XING Huan-chun;CAO Wen-bin;ZHANG Yi;WANG Yong-an;YANG Jun(State Key Laboratory of Highly Toxic Substances for Anti-Drug Research,Institute of Pharmacology and Toxicology,Academy of Military Medical Sciences,Beijing 100850,China;Regiment 95865 of PLA,Beijing 102200,China;Hospital 901 of the Joint Logistic Support Force of PLA,Hefei 230009,China)

机构地区：[1]军事科学院军事医学研究院毒物药物研究所,高毒物质对抗药物研究国家重点实验室,北京100850 [2]中国人民解放军95865部队,北京102200 [3]中国人民解放军联勤保障部队第九零一医院,安徽合肥230009

出　　处：《中国药理学与毒理学杂志》2023年第4期274-280,共7页Chinese Journal of Pharmacology and Toxicology

基　　金：北京市科技新星项目(Z191100001119105)。

摘　　要：目的评价梭曼、沙林、塔崩和维埃克斯(VX)4种有机磷化合物的体外稳定性及小鼠染毒后毒剂分子在血液中活性持续时间,为有机磷化合物中毒救治中药物的使用频率和给药时间等提供参考和指导。方法(1)有机磷化合物体外稳定性评价。梭曼、沙林和VX分别用无菌水稀释1×105倍,塔崩用无菌水稀释1×104倍,将以上稀释好的4种有机磷化合物再分别用PBS稀释100倍后置37℃恒温培养箱孵育,分别在0,10,20,40,90,120和240 min取出一定量有机磷化合物与乙酰胆碱酯酶(AChE)溶液0.2 g·L-1反应,采用Ellman法测定AChE活性,并计算酶活抑制率。(2)有机磷化合物染毒小鼠后血液分离物对AChE活性抑制作用。将4种有机磷化合物配置成不同浓度梯度后对KM小鼠进行腹部皮下染毒,以组内小鼠未出现死亡时的最高使用剂量为随后建模的动物染毒剂量,在该剂量染毒后分别于0,10,20,40,90,120和240 min取全血,采用超滤离心法分离各时间点体内残留的有机磷化合物并采用Ellman法测定其对AChE活性抑制率。结果(1)在体外稳定性实验中,各时间点梭曼的AChE活性抑制率为90%~95%,与0 min组相比无明显差异;沙林的AChE活性抑制率为20%~70%,与0 min组相比,40 min时酶活抑制率开始明显下降(P<0.05),直到240 min时急剧下降至0 min组的约50%(P<0.01);塔崩的AChE活性抑制率为5%~50%,从10 min起明显下降(P<0.05),随时间延长逐渐降低,240 min时衰减至0 min组的约10%(P<0.01);VX的AChE活性抑制率为80%~90%,整个时段酶活抑制率未见明显变化。(2)在体内实验中,确定梭曼140μg·kg^(-1)、沙林220μg·kg^(-1)、塔崩300μg·kg^(-1)和VX 14μg·kg^(-1)作为KM小鼠染毒最佳剂量。与0 min组相比,梭曼染毒40~90 min内,血液分离物对ACh E的活性抑制率显著升高(P<0.05);沙林染毒20后直到240 min,血液分离物对AChE的活性抑制率均显著升高(P<0.05,P<0.01);塔崩染毒后20 min和40 min,血液分离物对OBJECTIVE To evaluate the stability of soman,sarin,tabun and VX in vitro and their duration of activity in vivo after mouse exposure so as to provide reference and guidance for the frequency and time of administration of drugs in the treatment of organophosphorus compound poisoning.METHODS①Evaluation of stability of organophosphates in vitro:soman,sarin and VX were diluted with sterile water by 1×105 times,respectively,and tabun by 1×104 times,Then,the four organophosphorus compounds were diluted 100 times with PBS,respectively,and placed in a 37℃incubator for 0,10,20,40,90,120,and 240 min.Samples at different time points reacted with known concentrations of acetylcholinesterase(AChE)solution(0.2 g·L-1).AChE activity was determined with the Ellman method.The inhibitory rate of organo⁃phosphorus compounds against AChE was calculated.②The inhibitory effect of blood isolates on AChE after exposure to organophosphorus compounds in vivo:a series of different concentrations of soman,sarin,tabun and VX was prepared,and their optimum exposure concentrations was explored in KM mice after abdominal subcutaneous injection.After 0,10,20,40,90,120 and 240 min of exposure,the whole blood was taken,and the residual organophosphorus compounds in the body at different time points were separated via ultrafiltration centrifugation.The isolates reacted with known concentrations of AChE solution.The activity of AChE was determined using the Ellman method and the inhibitory rate was calculated.RESULTS The inhibitory rate of soman on AChE activity was 90-95%in vitro,and there was no statistically significant difference compared with 0 min.The inhibitory rate of sarin on AChE activity ranged from 20%to 70%in vitro.The inhibition lasted for some time before it decreased gradually.Between 0 and 40 min,the inhibitory rate of sarin on AChE activity hardly changed,but began to decrease at 40 min,and the difference was statistically significant(P<0.05).This inhibitory rate also decreased significantly between 90 min and 240 min(P<0

关 键 词：有机磷化合物 胆碱酯酶 酶活性 

分 类 号：R99[医药卫生—毒理学]