单独或与鲨鱼软骨提取物联合对伊朗利什曼原虫标准菌株（MRHO/IR/75/ER）的强效抗利什曼原虫活性的体外评估

In vitro Evaluation of the Potent Antileishmanial Activity of Alone or in Combination with Shark Cartilage Extract Against the Standard Iranian Strain of (MRHO/IR/75/ER).

作者信息

Saraei Shaylin, Soozangar Narges, Miran Mansour, Ghaffarifar Fatemeh, Mohammadi-Ghalehbin Behnam, Molaei Soheila, Asfaram Shabnam

机构信息

Students Research Committee, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran.

Zoonoses Research Center, Ardabil University of Medical Sciences, Ardabil, Iran.

出版信息

Iran J Pharm Res. 2023 Jul 6;22(1):e136173. doi: 10.5812/ijpr-136173. eCollection 2023 Jan-Dec.

DOI:10.5812/ijpr-136173

PMID:38116562

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10728847/

Abstract

BACKGROUND

The available drugs for the treatment of leishmaniasis are highly toxic and extremely expensive, with low efficiency; therefore, the development of effective therapeutic compounds is essential.

OBJECTIVES

The present study aimed to explore the antileishmanial effects of ethyl acetate extract, methanol extract, and fractions 1-4 (F1-F4) of , alone or in combination with shark cartilage extract (ShCE), on in vitro.

METHODS

In this study, ethyl acetate, methanol, and n-hexane extracts were extracted from the aerial roots of by the maceration method. The silica gel column chromatography was used to separate n-hexane extracts at varying polarities (F1-F4 fractions). Subsequently, the effects of extracts and fractions against promastigotes were assessed by the parasite counting method microscopic inhibition test and MTT assay. Besides, their effects on the infected macrophage cells and the number of amastigotes were investigated. Cytotoxicity was evaluated in non-infected J774A.1 macrophage cells. Finally, apoptosis induction of promastigotes, including infected and non-infected macrophages, was evaluated.

RESULTS

The results indicated the highly potent activity of extracts and F1-F4 fractions, alone or in combination with ShCE, against promastigotes and amastigotes in a dose-dependent manner (P < 0.05). The F1 fraction and methanol extract showed markedly higher toxicity compared to the other extracts and fractions, with 50% inhibitory concentrations (IC50/72h) of 2.4 ± 0.29 and 2.9 ± 0.55 µg/mL against promastigotes and 1.79 ± 0.27 µg/mL and 1.39 ± 0.27 µg/mL against amastigotes (P < 0.001). Moreover, they had a high selectivity index (SI) due to the low toxicity of macrophages (P < 0.0001). The results of flow cytometry indicated that the percentages of apoptotic promastigote cells in contact with IC50 concentrations of F1 and methanol extract alone after 72 h were 43.83 and 43.93%, as well as 78.4%, and 65.45% for their combination with ShCE, respectively.Also, apoptosis of infected macrophages induced by F1 and methanol extracts was estimated at 68.5% and 83.7%, respectively.

CONCLUSIONS

In this study, the F1 fraction and methanol extract of showed potent efficacy against , associated with low toxicity and apoptosis induction. Therefore, they can be promising therapeutic candidates in future animal and even human studies.

摘要

背景

现有的治疗利什曼病的药物毒性高、价格昂贵且效率低；因此，开发有效的治疗化合物至关重要。

目的

本研究旨在探讨的乙酸乙酯提取物、甲醇提取物以及馏分1 - 4（F1 - F4）单独或与鲨鱼软骨提取物（ShCE）联合对的体外抗利什曼原虫作用。

方法

在本研究中，采用浸渍法从的气生根中提取乙酸乙酯、甲醇和正己烷提取物。使用硅胶柱色谱法分离不同极性的正己烷提取物（F1 - F4馏分）。随后，通过寄生虫计数法、显微镜抑制试验和MTT试验评估提取物和馏分对前鞭毛体的作用。此外，研究了它们对感染巨噬细胞和无鞭毛体数量的影响。在未感染的J774A.1巨噬细胞中评估细胞毒性。最后，评估前鞭毛体（包括感染和未感染的巨噬细胞）的凋亡诱导情况。

结果

结果表明，的提取物和F1 - F4馏分单独或与ShCE联合对的前鞭毛体和无鞭毛体具有高效活性，且呈剂量依赖性（P < 0.05）。与其他提取物和馏分相比，F1馏分和甲醇提取物显示出明显更高的毒性，对前鞭毛体的50%抑制浓度（IC50/72h）分别为2.4 ± 0.29和2.9 ± 0.55 µg/mL，对无鞭毛体的抑制浓度分别为1.79 ± 0.27 µg/mL和1.39 ± 0.27 µg/mL（P < 0.001）。此外，由于对巨噬细胞毒性低（P < 0.0001），它们具有高选择性指数（SI）。流式细胞术结果表明，在72小时后，单独接触IC50浓度的F1和甲醇提取物的凋亡前鞭毛体细胞百分比分别为43.83%和43.93%，与ShCE联合时分别为78.4%和65.45%。此外，F1和甲醇提取物诱导的感染巨噬细胞凋亡分别估计为68.5%和83.7%。