Molaie Soheila, Ghaffarifar Fatemeh, Hasan Zuheir Mohammad, Dalimi Abdolhosein
Department of Parasitology , Faculty of Medical Sciences, Tarbiat Modares University, Tehran, I.R.Iran.
Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, I.R.Iran.
Iran J Pharm Res. 2019 Spring;18(2):887-902. doi: 10.22037/ijpr.2019.1100656.
In this study we examined enhancement effects of Artemisinin plus Glucantime and shark cartilage extract on promastigotes and amastigotes of in condition.The toxicity of artemisinin, glucantime, and shark cartilage extract on the promastigotes and amastigote-infected macrophages was evaluated using MTT assay. The role of these drugs inducing apoptosis in promastigotes, un- infected, and parasite- infected macrophages was also studied. Using promastigote assay, IC50 values of artemisinin and glucantime as standalone drugs as well as in combination were obtained to be 50, 400, and 100µg/mL respectively. The flow cytometry analysis of apoptotic promastigotes stained with Annexin-V FITC staining showed that artemisinin, glucantime, artemisinin plus glucantime, artemisinin plus shark cartilage extract, and shark cartilage extract alone applied at their IC50 concentrations resulted in 53.5%, 73.92%, 64.46%, 49.9%, and 47.34% apoptosis respectively. The results of MTT assay indicated that cytotoxicity of artemisinin, glucantime, artemisinin plus glucantime, shark cartilage plus artemisinin, and shark cartilage in infected macrophages after 72h was 75%, 84%, 82%, 30%, and 3% respectively. In un- infected macrophages, cytotoxicity of Artemisinin, Glucantime, Artemisinin plus Glucantime and shark cartilage was 15%, 31%, 21%, 2%, and 0% respectively.This study suggests that artemisinin, glucantime, artemisinin plus glucantime, and shark cartilage extract have significant killing effects on promastigotes and amastigotes. Also, it proved that artimisinin alone and in combination with glucantime and shark cartilage extract has little toxic effect on macrophages, but could induce apoptosis in promastigotes and amastigote-infected macrophages. Thus, these chemicals can be used as alternative drugs for studies.
在本研究中，我们检测了青蒿素加葡糖胺锑钠以及鲨鱼软骨提取物在体外条件下对前鞭毛体和无鞭毛体的增强作用。使用MTT法评估青蒿素、葡糖胺锑钠和鲨鱼软骨提取物对前鞭毛体以及无鞭毛体感染的巨噬细胞的毒性。还研究了这些药物在诱导前鞭毛体、未感染的巨噬细胞以及寄生虫感染的巨噬细胞凋亡中的作用。通过前鞭毛体试验，单独使用青蒿素和葡糖胺锑钠以及联合使用时的IC50值分别为50、400和100μg/mL。用膜联蛋白-V FITC染色对凋亡前鞭毛体进行流式细胞术分析表明，以其IC50浓度单独应用青蒿素、葡糖胺锑钠、青蒿素加葡糖胺锑钠、青蒿素加鲨鱼软骨提取物以及鲨鱼软骨提取物时，凋亡率分别为53.5%、73.92%、64.46%、49.9%和47.34%。MTT试验结果表明，72小时后青蒿素、葡糖胺锑钠、青蒿素加葡糖胺锑钠、鲨鱼软骨加青蒿素以及鲨鱼软骨在感染巨噬细胞中的细胞毒性分别为75%、84%、82%、30%和3%。在未感染的巨噬细胞中，青蒿素、葡糖胺锑钠、青蒿素加葡糖胺锑钠以及鲨鱼软骨的细胞毒性分别为15%、31%、21%、2%和0%。本研究表明，青蒿素、葡糖胺锑钠、青蒿素加葡糖胺锑钠以及鲨鱼软骨提取物对前鞭毛体和无鞭毛体具有显著的杀伤作用。此外，还证明单独使用青蒿素以及与葡糖胺锑钠和鲨鱼软骨提取物联合使用时对巨噬细胞几乎没有毒性作用，但可诱导前鞭毛体和无鞭毛体感染的巨噬细胞凋亡。因此，这些化学物质可作为用于相关研究的替代药物。
