Rezaei Riabi Tahereh, Sharifi Iraj, Miramin Mohammadi Akram, Khamesipour Ali, Hakimi Parizi Maryam
Leishmaniasis Research Center, Kerman University of Medical Sciences, Kerman, Iran.
Center for Research &Training in Skin Diseases & Leprosy, Tehran University of Medical Sciences, Tehran, Iran.
Iran J Parasitol. 2013 Jul;8(3):396-401.
Pentavalent antimonials are still the first choice treatment for leishmaniasis, but with low efficacy and resistance is emerging. In the present study, the effect of meglumine antimoniate (MA, Glucantime) combined with paromomycin, miltefosine or allopurinol on in vitro susceptibility of Leishmania tropica resistant isolate was evaluated.
The drugs were obtained from commercial sources and diluents of each drug in medium were prepared on the day of experiment. J774 A.1 murine macrophage cell lines were attached to the cultured on slide and incubated at 37 °C with 5% CO2 for 24 h. Then the stationary phase promastigotes were added to the cells and after 4 hrs of incubation different concentrations of MA, paromomycin, miltefosine or allopurinol were added and incubated for an additional of 72 h. Then the slides were dried and fixed with methanol, stained by Giemsa and studied under a light microscope. Drug activity was evaluated by assessing the macrophage infection rate and the number of amastigotes per infected macrophage was done by examining 100 macrophages. The experiment was done in triplicates.
Various concentrations of MA along with paromomycin, miltefosine or allopurinol significantly inhibited (P<0.01) the proliferation of L. tropica amastigote stage in the macrophage cell line as compared with MA alone or positive control.
Combination of Glucantime with paromomycin, miltefosine or allopurinol showed a synergistic effect on the clinical isolate of L. tropica in vitro. Use of combination therapy is a new hope and a logical basis for therapy of the patients with cutaneous leishmaniasis. Further investigations are needed to evaluate the therapeutic effects of these drugs on the CL patients.
五价锑制剂仍然是利什曼病的首选治疗药物，但疗效较低且耐药性正在出现。在本研究中，评估了葡甲胺锑（MA，葡醛锑钠）联合巴龙霉素、米替福新或别嘌呤醇对热带利什曼原虫耐药分离株体外敏感性的影响。
药物购自商业渠道，每种药物在培养基中的稀释液在实验当天配制。将J774 A.1小鼠巨噬细胞系贴壁于载玻片上培养，在37℃、5%二氧化碳条件下孵育24小时。然后将稳定期前鞭毛体加入细胞中，孵育4小时后加入不同浓度的MA、巴龙霉素、米替福新或别嘌呤醇，再孵育72小时。然后将载玻片干燥，用甲醇固定，吉姆萨染色，在光学显微镜下观察。通过评估巨噬细胞感染率来评价药物活性，通过检查100个巨噬细胞来确定每个感染巨噬细胞内无鞭毛体的数量。实验重复三次。
与单独使用MA或阳性对照相比，不同浓度的MA联合巴龙霉素、米替福新或别嘌呤醇均显著抑制（P<0.01）巨噬细胞系中热带利什曼原虫无鞭毛体阶段的增殖。
葡醛锑钠联合巴龙霉素、米替福新或别嘌呤醇在体外对热带利什曼原虫临床分离株显示出协同作用。联合疗法的应用为皮肤利什曼病患者的治疗带来了新希望和合理依据。需要进一步研究来评估这些药物对皮肤利什曼病患者的治疗效果。
