Abstract：
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.
摘要：
棘阿米巴是自由生活的变形虫，是角膜炎和肉芽肿性阿米巴脑炎的病原体。Alpha-Mangostin（AMS）是一种重要的黄吨酮，具有广泛的生物活性。这里,在体外评估了α-Mangostin及其银纳米缀合物（AMS-AgNPs）对致病性A.castellanii滋养体和囊肿的抗阿米巴活性。杀菌试验表明，AMS和AMS-AgNPs均剂量依赖性地抑制了A.castellanii的活力，IC50为88.5±2.04和20.2±2.17μM，分别。两种制剂均抑制卡氏A.castellanii介导的人角质形成细胞的细胞致病性。功能测定显示，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。全转录组测序显示了826个基因的表达，447个基因上调，379个基因在治疗后下调。京都基因和基因组百科全书分析表明，大多数基因与细胞凋亡有关，自噬,RAP1、AGE-RAGE和催产素信号通路。七个基因（PTEN，H3,ARIH1,SDR16C5,PFN,glnaGLUL,和SRX1）被鉴定为体外分子作用模式的最重要（Log2（FC）值4）。需要使用AMS和纳米缀合物的未来体内研究来实现这项工作的临床潜力。
