目的 观察花椒乙醇提取物对体外培养的蓝氏贾第鞭毛虫（简称贾第虫）滋养体的抑制作用和结构变化,探讨花椒体外抗贾第虫的作用效果。 方法 将7.5 × 10 ⁵~2.5 × 10 ⁶个/ml贾第虫滋养体分别给予不同浓度的花椒乙醇提取物和阿苯达唑,花椒乙醇提取物组的终浓度分别为0.05、0.50、5.00、50.00、250.00 mg/ml,阿苯达唑组终浓度分别为0.04、0.40、4.00、40.00、200.00 mg/ml,同时设空白对照组。采用改良TYI-S-33培养基37 ℃厌氧培养24 h,计数并观察两种药液对贾第虫的作用效果,每个浓度重复3次。采用SPSS 22.0软件计算花椒乙醇提取物和阿苯达唑的半抑制浓度（IC₅₀）。取IC₅₀浓度的花椒乙醇提取物加入体外培养的贾第虫滋养体中,37 ℃分别培养1、3、7、15、24 h后观察虫体活力、形态结构变化,计算虫体致死率,另设空白对照组,每个时间点重复3次。扫描电镜观察虫体形态变化。 结果 空白对照组贾第虫生长状态良好,虫体饱满并且数量多,而不同浓度花椒乙醇提取物和阿苯达唑组的贾第虫生长均受到抑制。与空白对照组比,花椒乙醇提取物浓度为5~250 mg/ml,阿苯达唑为0.04~200 mg/ml时,虫体形态差异均有统计学意义（P < 0.05）。SPSS统计得到花椒乙醇提取物的IC ₅₀为4.57 mg/ml,阿苯达唑的IC₅₀为0.07 mg/ml。当给予两种药液IC₅₀浓度时,随着作用时间的延长,对虫体的抑制作用逐渐增强。给药后培养7 h,致死率均超过50%;给药后培养15 h,致死率均超过70%;给药后培养24 h,花椒乙醇提取物组致死率达（80.03 ± 3.62）%;阿苯达唑组致死率达（91.70 ± 4.91）%,与空白对照组相比差异均有统计学意义（ P < 0.05）。扫描电镜观察发现,给予花椒乙醇提取物后7 h时,虫体变形,表面凹凸不平,但鞭毛还清晰可见;给药后12 h以上,整个滋养体停止生长,鞭毛受到明显损伤,腹吸盘表面细胞膜破裂,细胞内容物外溢,细胞失去支撑而变得扁平;给药后24 h,虫体表面破损严重,虫体表膜皱缩,进而导致虫体立体结构改变,胞质耗空,最后仅剩下躯壳而死亡。 结论 花椒乙醇提取物对体外培养的贾第虫生长有一定的抑制作用,并且随着药液浓度的增高,对虫体的抑制作用更加显著。

Objective To investigate the effect of Zanthoxylum bungeanum against Giardia duodenalis by observing inhibitory effect and structural changes on the parasite in vitro cultures by Z. bungeanum ethanol extracts. Methods Giardia strophozoites at the concentration of 7.5×10⁵-2.5×10⁶/ml in culture was treated with different concentrations of Z. bungeanum Maxim solution (0.05, 0.500, 5.00, 50.00, 250.00 mg/ml) and albendazole solution at final concentrations of 0.04, 0.40, 4.00, 40.00, and 200.00 mg/ml, respectively. A blank treatment group served as control. After anaerobic culture at 37 ℃ for 24 h using modified TYI-S-33 medium, the inhibitory effects of the two testing reagents on the growth of G. duodenalis were observed and counted. Each concentration was tested in triplicates. The half maximal inhibitory concentration (IC₅₀) values of Z. bungeanum extracts and albendazole were calculated using the SPSS 22.0 software. Then, the trophozoites of G. duodenalis were treated in vitro with the Z. bungeanum extracts at the IC₅₀ concentration for 1, 3, 7, 15 and 24 h at 37 ℃. Blank control group was set for the study. The vitality, morphological and structural changes were observed, and fatality rate of the trophozoites was calculated (repeated 3 times at each time point), and the morphological changes were observed by scanning electron microscopy.Results G. duodenalis in the blank control group showed normal growth, fullness appearance with increased number, while the Z. bungeanum Maxim at different concentrations and albendazole solution suppressed the growth of parasites. Compared with the blank control group, tthe trophozoites treated with Z. bungeanum extracts (5-250 mg/ml) and albendazole solution (0.04-200 mg/ml) showed significant morphological changes (P < 0.05). The IC ₅₀ values of Z. bungeanum extracts and albendazole were 4.57 mg/ml and 0.07 mg/ml, respectively. At the IC₅₀ concentrations, suppressive effect on the parasites was increased with longer exposure time. Exposed to the reagents for 7 h, the parasite fatality was higher than 50%, and the exposure for 15 h resulted in a fatality rate higher than 70%. The fatality reached (80.03 ± 3.62)% and (91.70 ± 4.91)% after 24 h treatment with Z. bungeanum extracts and albendazole, respectively (P < 0.05 compared with the control group). Electron microscopy found after treatment with Z. bungeanum extract for 7 h, the parasite deformed with irregular surface, but the flagella remained clearly visible; upon treatment for > 12 h, the growth of the trophozoites ceased, flagella were obviously damaged, surface cytomembrane of ventral acetabula were broken, cell content spilled, cells flattened due to losing skeleton brace; after treatment for 24, worm surface were seriously damaged, worm epithelial membrane shrank, leading to changes in worm stereo structure, and eventually leaving only the worm outer form due to depleted cytoplasm. Conclusion The Z. bungeanum extract has certain suppressive effect on the growth of G. duodenalis, and the suppressive effect on the worms was enhanced with the increasing concentration of the testing extracts.