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目的 评价双氢青蒿素对棘阿米巴的体外抑制作用.设计 实验研究.研究对象 临床患者角膜分离的阿米巴虫株.方法 体外培养阿米巴虫株,加入不同稀释浓度(3.2％、1.6％、0.8％、0.4％、0.2％、0.1％、0.05％、0.025％和0.0125％)的双氢青蒿素(DHA)共同孵育,同时以洗必泰(CHG)和聚六甲基双胍(PHMB)做对照药物,通过转种阿米巴培养基和LIVE/DEAD染色观察药物对阿米巴活性的抑制作用.主要指标 光学显微镜观察药物作用后虫体形态变化;LIVE/DEAD染色后,荧光显微镜下阿米巴活性情况,计算最小杀包囊浓度(MCC)和最小杀滋养体浓度(MTAC)值,以及各MTAC和MCC水平下阿米巴虫株的累计分布率,即MTAC50或MCC50和MTAC90或MCC90.结果 药物抑制后的阿米巴包囊干瘪皱缩,LIVE/DEAD法观察,受抑制的包囊呈红色,无活性.DHA组MTAC和MCC值范围均为0.05％～1.6％.MTAC50和MCC50值均为0.4％,MTAC90和MCC90值均为1.6％;CHG组MTAC值范围0.0125％～1.6％,MCC值范围0.0125％～0.8％,MTAC50和MCC50值分别为0.05％和0.1％,MTAC90和MCC90值均为0.8％.PHMB组MTAC值范围0.00625％～0.4％,MCC值范围0.00625％～0.2％,MTAC50和MCC50值均为0.025％,MTAC90和MCC90值均为0.2％.三组药物对不同阿米巴菌株的MCC和MTAC相比较均无统计学差异(P=0.342、0.459、0.469).三组MTAC值总体比较DHA组最高,CHG组次之,PHMB组最低(F=3.813,P=0.035).其中,DHA组高于PHMB组(P=0.011);DHA组和CHG组以及CHG组和PHMB组比较差异无统计学意义(P=0.105和0.297).三种药物组的MCC值总体比较差异有统计学意义(F=6.672,P=0.004),DHA组比CHG组高,DHA组比PHMB组高(P=0.017和0.001);CHG组与PHMB组无差异(P=0.325).结论 双氢青蒿素在体外对棘阿米巴有抑制作用,其临床治疗阿米巴性角膜炎的作用仍待验证.","Objective To evaluate the inhibitory effect of dihydroartemisinine (DHA) on acanthamoeba in vitro.Design Experimental study.Participants Acanthamoeba isolated from patients coea.Methods Culture acanthamoeba in vitro and add different concentrations of dihydroartemisinin (3.2％,1.6％,0.8％,0.4％,0.2％,0.1％,0.05％,0.025％,and 0.0125％).They were incubated together and used chlorhexidine (CHG) and PHMB as control drugs.The inhibitory effect of drugs on acanthamoeba were observed through turing acanthamoeba medium and LIVE/DEAD staining.Main Outcome Measures Morphological changes of mites under optical microscope after drug action.Activity of acanthamoeba under fluorescence microscope after LIVE/DEAD staining.Measure minimum cysticidal concentration (MCC) and minimum trophozoite amoebicidal concentration (MTAC).The cumulative distribution rate of acanthamoeba under MTAC and MCC levels was MTAC50 and MCC50 or MTAC90 and MCC90.Results The range of MTAC and MCC in DHA group were both 0.05％～0.6％;MTAC50 and MCC50 were 0.4％ both,while both MTAC90 and MCC90 were 1.6％.The MTAC range of CHG group was 0.0125％～1.6％;MCC range was 0.0125％～0.8％,MTAC50 and MCC50 were 0.05％ and 0.1％;MTAC90 and MCC90 were both 0.8％.The MTAC of PHMB group was 0.00625％～0.4％;MCC range was 0.00625％～0.2％;MTAC50 and MCC50 were both 0.025％;MTAC90 and MCC90 were both 0.2％.There was no statistically significant difference between MCC and MTAC in the three groups for different strains of acanthamoeba (P=0.342,0.459,0.469).The difference of the MTAC among DHA,CHG and PHMB group was statistically significant (F=3.813,P=0.035).The MTAC of DHA group was higher than PHMB group(P=0.011).There was no statistically significant difference of the MTAC between the DHA and CHG group as well as CHG and PHMB group (P=0.105,0.297).The difference of the MCC among DHA,CHG and PHMB group was statistically significant difference (F=6.672,P=0.004).The MCC of DHA group was higher than CHG group,and DHA group was higher than PHMB group (P=0.017,0.001).There was no significant difference of the MCC between CHG and PHMB group (P=0.325).Conclusion The inhibitory effect of the dihydroartemisinine on acanthamoeba is observed in vitro,and the effect on acanthamoeba keratitis in clinic remains to be further studied.