Acanthamoeba are free living amoebae that are the causative agent of keratitis and granulomatous amoebic encephalitis. Alpha-Mangostin (AMS) is a significant xanthone; that demonstrates a wide range of biological activities. Here, the anti-amoebic activity of α-Mangostin and its silver nano conjugates (AMS-AgNPs) were evaluated against pathogenic A. castellanii trophozoites and cysts in vitro. Amoebicidal assays showed that both AMS and AMS-AgNPs inhibited the viability of A. castellanii dose-dependently, with an IC₅₀ of 88.5 ± 2.04 and 20.2 ± 2.17 μM, respectively. Both formulations inhibited A. castellanii-mediated human keratinocyte cell cytopathogenicity. Functional assays showed that both samples caused apoptosis through the mitochondrial pathway and reduced mitochondrial membrane potential and ATP production, while increasing reactive oxygen species (ROS) and nicotinamide adenine dinucleotide phosphate (NADPH) cytochrome-c reductase in the cytosol. Whole transcriptome sequencing of A. castellanii showed the expression of 826 genes, with 447 genes being up-regulated and 379 genes being down-regulated post treatment. The Kyoto Encyclopedia of Genes and Genomes analysis showed that the majority of genes were linked to apoptosis, autophagy, RAP1, AGE-RAGE and oxytocin signalling pathways. Seven genes (PTEN, H3, ARIH1, SDR16C5, PFN, glnA GLUL, and SRX1) were identified as the most significant (Log2 (FC) value 4) for molecular mode of action in vitro. Future in vivo studies with AMS and nanoconjugates are needed to realize the clinical potential of this work.

Graphical abstract

中文翻译：

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Alpha-Mangostin 及其纳米缀合物诱导 T4 基因型卡斯氏棘阿米巴细胞程序性死亡

棘阿米巴是自由生活的阿米巴，是角膜炎和肉芽肿性阿米巴脑炎的病原体。Alpha-Mangostin (AMS) 是一种重要的氧杂蒽酮；这表明了广泛的生物活性。在此，我们在体外评估了 α-Mangostin 及其银纳米缀合物 (AMS-AgNP) 对致病性A.castellanii滋养体和包囊的抗阿米巴活性。杀阿米巴试验表明，AMS 和 AMS-AgNP 均以剂量依赖性方式抑制A.castellanii的活力，IC ₅₀分别为 88.5 ± 2.04 和 20.2 ± 2.17 μM。两种制剂均抑制卡斯氏放线菌介导的人角质形成细胞的细胞致病性。功能分析表明，两种样品均通过线粒体途径引起细胞凋亡，并降低线粒体膜电位和 ATP 产生，同时增加细胞质中的活性氧 (ROS) 和烟酰胺腺嘌呤二核苷酸磷酸 (NADPH) 细胞色素 c还原酶。A.castellanii的全转录组测序显示826个基因的表达，其中447个基因在处理后上调，379个基因下调。京都基因和基因组百科全书分析显示，大多数基因与细胞凋亡、自噬、RAP1、AGE-RAGE 和催产素信号通路有关。七个基因（PTEN、H3、 ARIH1 、SDR16C5、PFN 、 glnA GLUL和SRX1）被确定为体外分子作用模式最显着（Log2 (FC) 值 4）。未来需要使用 AMS 和纳米缀合物进行体内研究，以实现这项工作的临床潜力。

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