Azas N, Di Giorgio C, Delmas F, Gasquet M, Timon-David P
Laboratoire de Parasitologie, Faculté de Pharmacie, Marseille, France.
Exp Parasitol. 1997 Sep;87(1):1-7. doi: 10.1006/expr.1997.4182.
The capacity of flow-cytometric techniques to detect drug-specific biochemical targets and side effects in Leishmania infantum promastigotes was estimated by assessing the effects of three antileishmanial drugs (pentamidine, allopurinol, and amphotericin B) on parasite metabolism. Cell cycle and total protein content were estimated by staining cells with propidium iodide and fluorescein isothiocyanate, nonprotein thiols were stained by mercury orange, and membrane potential was measured by the accumulation of 3,3'-dipenthyloxacarbocyanine iodide inside the cell. Results showed that dynamic studies in parasites treated with subtoxic concentrations of drugs allowed the detection of drug-specific targets: pentamidine primarily affected nonprotein thiol contents and DNA synthesis, allopurinol primarily affected intracellular protein contents, and amphotericin B primarily affected membrane potential. Moreover, the assessment of cellular functions in parasites treated with increasing concentrations of drugs certified the capacity of these techniques to establish dose-response curves and to permit the detection of side effects.
通过评估三种抗利什曼原虫药物（喷他脒、别嘌呤醇和两性霉素B）对寄生虫代谢的影响，估计了流式细胞术检测婴儿利什曼原虫前鞭毛体中药物特异性生化靶点和副作用的能力。用碘化丙啶和异硫氰酸荧光素对细胞进行染色来估计细胞周期和总蛋白含量，用汞橙对非蛋白硫醇进行染色，通过3,3'-二戊基氧杂羰花青碘化物在细胞内的积累来测量膜电位。结果表明，对用亚毒性浓度药物处理的寄生虫进行动态研究能够检测到药物特异性靶点：喷他脒主要影响非蛋白硫醇含量和DNA合成，别嘌呤醇主要影响细胞内蛋白质含量，两性霉素B主要影响膜电位。此外，对用浓度不断增加的药物处理的寄生虫的细胞功能评估证实了这些技术建立剂量反应曲线以及检测副作用的能力。
