目的 评价抗寄生虫中药水提物和有机溶剂粗提物体外对细粒棘球蚴的作用效果。方法 从感染绵羊肝脏细粒棘球蚴包囊中分离原头节，体外培养24 h后，取活性大于95%的原头节（100 μl，约100个）加入96孔板，分别加入终浓度为0.8 mg/ml（低浓度组）、1.6 mg/ml（中浓度组）、3.2 mg/ml（高浓度组）的鸦胆子、苦楝皮、辣蓼、石榴皮、使君子、槟榔、贯众、鱼藤、马蔺子、五倍子和仙鹤草等11种抗寄生虫中药的水提物，体外作用72 h后，伊红染色法检测原头节活性，倒置显微镜观察下观察原头节的形态，计算死亡率；同时设相同浓度的阿苯达唑组和空白对照组。从具有体外抗原头节作用的中药中提取黄酮、多糖、皂苷和生物碱等粗提物，分别以终浓度1.00 mg/ml体外作用于原头节72 h后，观察原头节形态，计算死亡率；同时设空白对照组。对体外抗原头节效果明显的粗提物进行液相色谱与串联质谱联用分析。组间比较采用单因素ANOVA分析，两两比较采用LSD-t检验。结果 中药水提物作用72 h后，鸦胆子、苦楝皮、辣蓼水提物高浓度组原头节边缘模糊，结构松散，基质溶解；其余8种中药水提物高浓度组原头节形态正常，各部分结构清晰；阿苯达唑组原头节生发层有轻微损伤；空白对照组原头节形态正常。体外培养72 h后，鸦胆子、苦楝皮和辣蓼水提物高浓度组原头节的死亡率分别为（99.63 ± 0.57）%、（90.89 ± 1.10）%和（51.93 ± 0.60）%，中浓度组分别为（85.97 ± 1.50）%、（81.14 ± 1.19）%、（42.46 ± 0.56）%，低浓度组分别为（78.34 ± 1.35）%、（77.27 ± 0.92）%、（36.66 ± 0.60）%，与空白对照组[（0.62 ± 0.51）%]相比差异均有统计学意义（F = 180 678.22、41 488.99、44 346.38，19 543.86、27 887.32、34 590.79，20 059.467、41 953.68、17 993.77，均P < 0.01），与阿苯达唑组[（30.03 ± 2.02）%]比较差异均有统计学意义（F = 6 585.72、4 210.84、646.46，2 956.80、2 849.68、210.83，2 365.92、2 712.28、58.12，均 P < 0.01）；其余8种中药水提物高、中、低浓度组原头节的死亡率与对照组比较差异均无统计学意义（均P > 0.05）。鸦胆子黄酮、苦楝皮黄酮、鸦胆子皂苷、辣蓼皂苷和辣蓼生物碱组原头节皱缩、结构松散、基质溶解，原头节死亡率分别为（98.33 ± 2.89）%、（96.67 ± 5.77）%、100%、（99.33 ± 1.15）%和（56.67 ± 2.11)%，与空白对照组[（10.33 ± 2.51）%]比较差异均有统计学意义（F = 1 584.00、563.71、3 808.47、3 099.52、14.65，均P < 0.01）；其余中药粗提物组原头节的死亡率与对照组比较差异均无统计学意义（均P > 0.05）。液相色谱与串联质谱联用分析结果显示，正、负离子模式下分别鉴定出741、398种代谢产物，其中正离子模式下从鸦胆子黄酮粗提物中鉴定出差异显著的黄酮类化合物4种，分别为漆黄素、5-甲基-7-甲氧基异黄酮、儿茶素和甜橙黄酮；负离子模式下鉴定出8种，分别为原花青素B2、槲皮素-3β-D-葡萄糖苷、枸桔苷、橙皮素、槲皮素、木犀草素、芹菜素和黄豆黄素。结论 鸦胆子、苦楝皮和辣蓼水提物，鸦胆子黄酮、苦楝皮黄酮、鸦胆子皂苷和辣蓼皂苷有明显体外抗细粒棘球蚴的作用，且效果优于阿苯达唑。

Objective To evaluate the in vitro effects of water extracts and organic solvent crude extracts of antiparasitic Chinese herb medicine against Echinococcus granulosus protoscolices Methods The protoscolices were isolated from the liver cyst of sheep infected with E. granulosus and cultured for 24 hours in vitro to collect protoscolices with vitality greater than 95%, which were transferred into 96 well plate (100 μl with 100 protoscolices). In the culture wells, the water extracts of 11 antiparasitic Chinese herb medicines, including Brucea javanica, Melia toosendan, Polygonum hydropiper, Pericarpium granati, Quisqualis linn, Areca catechu, Cyrtomium fortunei, Derris trifoliata, Semen iridis, Galla chinensis and Agrimonia pilosa, were applied at final concentrations of 0.8 mg/ml (low concentration group), 1.6 mg/ml (medium concentration group), and 3.2 mg/ml (high concentration group), respectively. After 72 hours of treatment, the protoscolices vitality was detected by eosin staining, and morphology was observed under inverted microscope to calculate the mortality rate. In addition, an albendazole group with the same concentration and a blank control group were designated. The crude herb extracts from flavonoids, polysaccharides, saponins and alkaloids showing apparent anti-protoscolices effect were cultured with protoscolices at a final concentration of 1.00 mg/ml for 72 hours. The morphology of protoscolices was observed to calculate the mortality rate. At the same time, blank control group was set up. Liquid chromatography and tandem mass spectrometry analysis were performed for those crude extracts showing significant anti-protoscolices effects. Single variant ANOVA analysis was used for inter-group comparisons, and LSD-t-test was used for pairwise comparison. Results After 72 hours of treatment with the traditional Chinese medicine water extracts, the protoscolices edges appeared blurred, the structure was loose, and the matrix was dissolved in the high-concentration groups with B. javanica, M. toosendan and P. hydropiper water extracts. The high concentration group of the other 8 Chinese herb medicine water extracts showed normal protoscolices morphology and clear structures. The albendazole group showed mild damage to the germinal layer of the original scalp. The blank control group had normal protoscolices morphology. After 72 hours of in vitro culturing, the protoscolices mortality rates in B. javanica, M. toosendan and P. hydropiper water extracts high concentration groups were (99.63 ± 0.57)%, (90.89 ± 1.10)% and (51.93 ± 0.60)%; (85.97 ± 1.50)%, (81.14 ± 1.19)% and (42.46 ± 0.56)% in the medium concentration groups, and (78.34 ± 1.35)%, (77.27 ± 0.92)% and (36.66 ± 0.60)% in the low concentration groups, respectively. The difference was statistically significant compared with the blank control group [(0.62 ± 0.51)%] (F = 180 678.22, 41 488.99, 44 346.38; 19 543.86, 27 887.32, 34 590.79; 20 059.467, 41 953.68, 17 993.77; P < 0.01 for all comparations) and compared with the albendazole group [(30.03 ± 2.02)%] (F = 6 585.72, 4 210.84, 646.46; 2 956.80, 2 849.68, 210.83; 2 365.92, 2 712.28, 58.12; P < 0.01 for all comparetions). There were no statistically significant differences in the mortality rates of the protoscolices between the control group and the high-concentration groups, the medium-concentration groups and the low-concentration groups of the other 8 Chinese herb medicine water extracts (all P > 0.05). The protoscolices in the groups of B. javanica flavonoids, M. toosendan flavonoids, B. javanica saponins, P. hydropipe saponins and P. hydropiper alkaloid showed wrinkling, loose structure, and matrix dissolution， and the mortality rates of the protoscolices in the groups were (98.33 ± 2.89)%, (96.67 ± 5.77)%, 100%, (99.33 ± 1.15)% and （56.67 ± 2.11)%, respectively. The differences were statistically significant compared with the blank control group (10.33 ± 2.51)% (F = 1 584.00, 563.71, 3 808.47, 3 099.52, 14.65; P < 0.01 for all comparisons). There was no statistically significant difference in the mortality rate of the protoscolices between the high-concentration group of other crude extracts of traditional Chinese herb medicine and the control group (P > 0.05 for all comparisons). The liquid chromatography combined with tandem mass spectrometry analysis showed that 741 and 398 metabolites were identified in the positive and negative ion modes, respectively. Among them, four flavonoids with significant differences were identified in the flavonoids crude extracts of B. javanica in the positive ion mode, namely lutein, 5-methyl-7-methoxyisoflavone, catechins, and sweet orange flavonoids. Eight species were identified in negative ion mode, namely proanthocyanidin B2 and quercetin B3 β-D-glucoside, citrin, hesperetin, quercetin, luteolin, apigenin, and daidzein. Conclusion The water extracts of B. javanica, M. toosendan, P. multiflorum, as well as B. javanica flavonoids, M. toosendan flavonoids, B. javanica saponins and P. hydropipe saponins reveal significant anti-echinococcus granulosus protoscolices in vitro, and their effects are superior to those of albendazole.