Aguirre-Cruz M L, Valadez-Salazar A, Muñoz O
Depto. de Biología Celular, Centro de Inv. y Estudios Avanzados, I.P.N., México, D.F.
Arch Invest Med (Mex). 1990;21 Suppl 1:23-6.
The chemotherapy of amebiasis includes different substances with intra and extraintestinal action. Nowadays, metronidazole is the election drug by its high efficacy and low toxicity. Some therapeutic failures with metronidazole in patients with invader amebiasis and some reports of resistance to it, of certain strains of Trichomonas vaginalis and species of Bacteroides have caused an increased interest to do new in vitro evaluation of this compound and to search of new antiamebic drugs. The object of this work was to standardize with our conditions a microtechnique to evaluate the sensibility to metronidazole, experimented by trophozoites of three amebic strains of different virulence (HK9:NIH, HM1:IMSS, HM3:IMSS) and to compare the effect exerted by the drug on them. We modified the technique proposed by Cedeño and Krogstad in 1983. First we read the results in a half of the time employed by them (24h) and after, we fixed the trophozoites with 0.25% glutaraldehyde before of make the counts of them. We compared the number of cells in microcultures of 3.2 x 10(4) trophozoites incubated during 24h with metronidazole (from 0.008 to 1 micrograms/ml) with the obtained in control microcultures. Each one of the strains showed to have a characteristic sensibility to the drug (p less than 0.001), which was directly proportional to the concentration. The middle effective dose of HK9 and HM1 strains was 0.3 microgram/ml and of 1 microgram/ml for HM3 strain. This technique seems to be reproducible and could be useful to quantify the in vitro activity of antiamebic agents.
阿米巴病的化疗包括具有肠内和肠外作用的不同药物。目前,甲硝唑因其高效低毒而成为首选药物。甲硝唑治疗侵袭性阿米巴病患者时出现的一些治疗失败情况,以及某些阴道毛滴虫菌株和拟杆菌属对其耐药的一些报道,使得人们对该化合物进行新的体外评估以及寻找新的抗阿米巴药物的兴趣增加。这项工作的目的是根据我们的条件规范一种微技术,以评估对甲硝唑的敏感性,该技术由三种不同毒力的阿米巴菌株(HK9:NIH、HM1:IMSS、HM3:IMSS)的滋养体进行实验,并比较该药物对它们的作用。我们修改了1983年Cedeño和Krogstad提出的技术。首先,我们在他们所用时间的一半(24小时)内读取结果,然后,在对滋养体进行计数之前,用0.25%的戊二醛固定滋养体。我们将3.2×10⁴个滋养体在24小时内与甲硝唑(浓度从0.008到1微克/毫升)一起孵育的微培养物中的细胞数量与对照微培养物中获得的细胞数量进行比较。每种菌株对该药物都表现出独特的敏感性(p<0.001),且与浓度成正比。HK9和HM1菌株的半数有效剂量为0.3微克/毫升,HM3菌株为1微克/毫升。该技术似乎具有可重复性,可用于量化抗阿米巴药物的体外活性。