他克林对多房棘球蚴抑制作用的实验研究  

Inhibitory efficacy of tacrine on Echinococcus multilocularis

作　　者：刘丛珊 尹建海 姚嘉青 张晶[2] 张皓冰 LIU Cong-shan;YIN Jian-hai;YAO Jia-qing;ZHANG Jing;ZHANG Hao-bing(National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention;Chinese Center for Tropical Diseases Researc;WHO Collaborating Centre for Tropical Disease;National Center for International Research on Tropical Diseases, Ministry of Science and Technolog;Key Laboratory of Parasite and Vector Biology, Ministry of Health, Shanghai 200025, China;2 Xinjiang Medical University, Urumqi 830011, Chin)

机构地区：[1]中国疾病预防控制中心寄生虫病预防控制所国家热带病研究中心世界卫生组织热带病合作中心科技部国家级热带病国际联合研究中心卫生部寄生虫病原与媒介生物学重点实验室,上海200025 [2]新疆医科大学,乌鲁木齐200025

出　　处：《中国寄生虫学与寄生虫病杂志》2018年第3期238-242,共5页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(No.81401691;81702030);上海市卫计委面上项目(No.201540170)~~

摘　　要：目的观察他克林体外抗多房棘球蚴的作用以及抗小鼠多房棘球蚴感染的疗效。方法在含50~200个多房棘球蚴原头节的96孔板中加入浓度为100.00、50.00、25.00、12.50和6.25μmol/L的他克林,给药后1、3、5和7 d用亚甲基蓝法测定原头节的活性。同时用细胞增殖-毒性检测(CCK-8法)测定他克林对3种正常宿主细胞(L929细胞、HK-2细胞和Chang liver)以及3种肿瘤细胞(A172细胞、A2058细胞和HCT-8细胞)的细胞毒性。40只感染多房棘球蚴6个月的BALB/c小鼠随机均分为4组,分别为30 mg/kg他克林组、15 mg/kg他克林组、25 mg/kg甲苯达唑组和1%西黄芪胶对照组。各组小鼠连续灌胃给药28 d,停药14 d后处死小鼠,剖取小鼠体内多房棘球蚴囊,称取囊重并计算囊重抑制率,采用SPSS 17.0中的非参检验法进行统计学分析。结果他克林体外对多房棘球蚴原头节的作用呈现明显的剂量效应关系,随着药物浓度和培养天数的增加,原头节的活性显著降低。100μmol/L的他克林作用3 d后,原头节全部死亡且形态发生强烈改变,难以观察到完整的超微结构。他克林作用7 d后,100.00、50.00、25.00、12.50和6.25μmol/L组的原头节死亡率分别为(100±0)%、(91.2±2.5)%、(80.3±5.1)%、(71.6±2.4)%和(51.7±2.9)%。他克林对正常宿主细胞活性影响较小,药物浓度增加至250.0μmol/L,对L929细胞、HK-2细胞和Chang liver细胞的活性抑制率分别为(27.6±4.7)%、(29.6±3.9)%和(26.9±2.1)%。但是对肿瘤细胞的细胞毒性较大,A172细胞、A2058细胞和HCT-8细胞的半数抑制浓度(Tox50)分别为(178.2±3.2)、(133.2±5.2)和(128.8±4.0)μmol/L。30 mg/kg他克林组、15 mg/kg他克林组和25 mg/kg甲苯达唑组囊重抑制率分别为-3.4%、9.4%和38.2%,上述各组小鼠体内多房棘球蚴囊重的减少与1%西黄芪胶组相比均无统计学意义(P>0.05)。4组小鼠在实验周期中分别有3、6、2和5只死亡,相较于其他组,25 mg/kg甲苯达唑和15 mg/kg他�Objective To evaluate the effects of tacrine on Echinococcus multilocularis in vitro and against E. multilocularis infection in mice. Methods Tacrine was added at 100.00, 50.00, 25.00, 12.50 and 6.25 μmol/L to each well of a 96-well plate containing 50-200 E. multilocularis protoscoleces. The methylene blue exclusion method was used to determine the viabilities of protoscoleces on days 1, 3, 5 and 7 after the addition. The cytotoxicity of tacrine on three noncarcinogenic cell types（L929 cells, HK-2 cells and Chang liver） and three tumor cell types（A172, A2058 and HCT-8 cells） was tested using the Cell Counting Kit-8（CCK-8） method. In another experiment, 40 BALB/c mice with 6 months of E. multilocularis infection were randomly divided into 4 groups： 30 mg/kg tacrine group, 15 mg/kg tacrine group, 25 mg/kg mebendazole group, and 1% tragacanth group as a control.All the mice received intragastric administration for 28 days. After 14 days of withdrawal, the mice were sacrificed,and cysts in the peritoneal cavity were isolated and weighed to calculate the reduction rate of cyst weight. Data were analyzed with the nonparametric test in SPSS 17.0. Results In vitro, tacrine showed an obvious dose-dependent effect on E. multilocularis protoscoleces. The viability of protoscoleces decreased with increased drug concentration and incubation time. After 3 days of exposure to 100 μmol/L tacrine, all the protoscoleces were dead,with dramatic morphological alterations. Intact parasites were hardly seen. After 7 days of tarcine treatment, the percentage dead of protoscoleces was（100 ± 0） %,（91.2 ± 2.5） %,（80.3 ± 5.1） %,（71.6 ± 2.4） % and（51.7 ±2.9） % in the 100.00, 50.00, 25.00, 12.50, and 6.25 μmol/L groups, respectively. The normal host cell types were less affected by tarcine at 250.0 μmol/L, as demonstrated by the（27.6 ± 4.7） %,（29.6 ± 3.9） % and（26.9 ±2.1） % inhibitory rate for L929 cells, HK-2 cells, and Chang liver cells, respectively. In contrast, tarcine exerted h

关 键 词：他克林 多房棘球蚴 原头节 细胞毒性 

分 类 号：R383.33[医药卫生—医学寄生虫学]