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Evaluation of curcumin and CM11 peptide alone and in combination against amastigote form of Iranian strain of L. major (MRHO/IR75/ER) in vitro
Experimental Parasitology ( IF 1.4 ) Pub Date : 2021-08-19 , DOI: 10.1016/j.exppara.2021.108151
G Aqeele 1 , P Shayan 2 , E Ebrahimzade Abkooh 3 , M Mohebali 4
Affiliation  

Curcumin (diferuloylmethane) is the main phytochemical of Curcuma longa Linn, an extract of the rhizome turmeric. For thousands of years, turmeric among other natural products has been used as a dietary spice and as a medicinal plant in Asian countries. The present study reports the leishmanicidal activity of curcumin in different concentrations (10 μM, 20 μM, 40 μM). It is also showing the effect of CM11 peptide (8 μM) alone and in combination with curcumin (10 and 20 μM) as a leishmanicidal drug. The experiments were performed with the amastigote form of Leishmania major (MRHO/IR/75/ER) in vitro and the leishmanicidal activity was analyzed after 12 and 24 h of incubation by Giemsa and DAPI staining. Further investigation was done by using semi-quantitative PCR with new designed common primer pair derived from an 18S rRNA gene belonging to the L. major and mouse, which amplified the above-mentioned gene segments simultaneously with different PCR product size. Our findings showed that curcumin had leishmanicidal activity in a dose and time-dependent manner and its lowest effective dose was at concentrations of 40 μM afetr12 h and 10 μM after 24 h. The IC50 value of curcumin against amastigote forms of L. major was 21.12 μM and 11.77 μM after 12 and 24 h, respectively. Treatment of amastigote form with CM11 (8 μM) alone and in combination with curcumin (10 μM and 20 μM) showed less leishmanicidal activity. Interestingly, CM11 in combination with curcumin (10 μM and 20 μM) had even less leishmanicidal effect compared to curcumin alone in the same concentrations (10 μM and 20 μM). The semi-quantitative PCR analysis confirmed the data achieved by Giemsa and DAPI staining and showed that curcumin reduced the PCR product derived from amastigote form in concentration and time-dependent manner compared to the genome of the host cells.



中文翻译:

在体外评估姜黄素和 CM11 肽单独和组合对抗伊朗大乳杆菌 (MRHO/IR75/ER) 无鞭毛体形式

姜黄素 (diferuloylmethane) 是Curcuma longa Linn的主要植物化学物质,姜黄是根茎姜黄的提取物。几千年来,姜黄和其他天然产品在亚洲国家一直被用作膳食香料和药用植物。本研究报告了不同浓度(10 μM、20 μM、40 μM)姜黄素的杀利什曼原虫活性。它还显示了 CM11 肽 (8 μM) 单独和与姜黄素(10 和 20 μM)组合作为利什曼原虫药物的作用。实验是用无鞭毛体形式的大利什曼原虫(MRHO/IR/75/ER)在体外进行的并在孵育 12 和 24 小时后通过吉姆萨和 DAPI 染色分析利什曼原虫的活性。通过使用半定量 PCR 和新设计的通用引物对进行进一步研究,该引物对来自属于L. Major和小鼠的 18S rRNA 基因,同时扩增上述基因片段,并具有不同的 PCR 产物大小。我们的研究结果表明,姜黄素具有剂量和时间依赖性的杀利什曼原虫活性,其最低有效剂量为 40 μM 后 12 小时和 10 μM 后 24 小时。姜黄素对L. Major 无鞭毛体形式的IC 5012 小时和 24 小时后分别为 21.12 μM 和 11.77 μM。用单独的 CM11 (8 μM) 和与姜黄素(10 μM 和 20 μM)组合处理无鞭毛体形式显示出较低的利什曼原虫活性。有趣的是,与相同浓度(10 μM 和 20 μM)的单独姜黄素相比,CM11 与姜黄素(10 μM 和 20 μM)组合的杀利什曼原虫作用甚至更小。半定量 PCR 分析证实了 Giemsa 和 DAPI 染色获得的数据,并表明与宿主细胞的基因组相比,姜黄素以浓度和时间依赖性方式减少了源自无鞭毛体形式的 PCR 产物。

更新日期:2021-08-25
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