H_2O_2体外抗棘阿米巴作用研究

目的检测H_2O_2的抗棘阿米巴(Acanthamoebaspp.)作用。方法自角膜炎患者角膜刮片分离获得棘阿米巴复合体(AcanthamoebaLugdunensis-Acanthamoebaquina)。于培养基(PYG)培养传代。实验前将棘阿米巴用新鲜的PYG培养1d使其活化,配成2.5×10~6/ml细胞悬液加入细胞培养板,实验组各孔分别加入不同浓度H_2O_2,对照组加等量PYG,28℃24h后实验组更换新鲜PYG继续培养3d。取细胞悬液滴片,瑞氏染色,观察细胞形态变化。用定量培养法作棘阿米巴生长曲线,观察其增殖速率。用四甲基偶氮唑盐(MTT)比色法,观察H_2O_2对棘阿米巴成活率的影响。用乳酸脱氢酶(LDH)测定法测定H_2O_2对棘阿米巴的损伤。结果棘阿米巴滋养体在0.125％H_2O_2作用下,不可逆转地成为包囊,20～120h增殖率为0;1％H_2O_2可使其破裂。结论H_2O_2具有较强的抗棘阿米巴作用,有可能成为预防棘阿米巴角膜炎的理想药物。

Study on the Effect of H_2O_2 Against Acanthamoeba in vitro

Objective To detect the effect of H_2O_2 on Acanthamoeba spp.. Methods By Wright's stain, quanti- tative culture, MTT assay and lactate dehydrogenase (LDH) assessment, the in fluence of H_2O_2 on the morphological fea- ture, proliferation speed and the survival rate of Acanthamoeba was tested. Results At low concentration of 0. 125%, H_2O_2 can force the Acanthamoeba trophozoites into cysts irreversibly, and inhibit its proliferation. 1% H_2O_2 can directly de- stroy Acanthamoeba trophozoites. Conclusion H_2O_2 is effective in destroying Acanthamoeba. It is possible to be used as an ideal reagent for the prevention of Acanthamoeba keratitis.