Mohammad Rahimi Hanieh, Mahdavi Fatemeh, Eslami Nasim, Nemati Sara, Mirjalali Hamed
Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
Acta Parasitol. 2025 Apr 12;70(2):89. doi: 10.1007/s11686-025-01024-z.
Hydatidosis is a zoonotic neglected disease caused by the larval stage of Echinococcus granulosus. Evidence suggests a communication between hydatid cyst (HC) and hosts via extracellular vesicles (EVs). However, a little is known about the communication between EVs derived from HC fluid (HCF) and host cells. The current study aimed to investigate the effect of HCF derived EVs on expression of fibrotic and anti-fibrotic miRNAs in THP-1 cell line.
In the current study, EVs were isolated using ultracentrifugation from wild-infected sheep HCF and characterized by western blot, electron microscope, and size distribution analysis. The effects of EVs on the expression levels of microRNAs (mir-16, mir-29a, and mir-155) involved in liver fibrosis were investigated using quantitative real-time PCR (qRT-PCR), 3 and 24 h after incubation.
Western blot analyses confirmed the expression of CD63 marker, while Calnexin and CD81 were absent in EVs samples. The SEM and morphology revealed round shape vesicles. The DLS analysis showed average size distribution 130.6 nm diameter. The expression levels of mir-16 and mir-29a were significantly upregulated after 3 h for 8.66 and 3.420, respectively, while they were significantly downregulated after 24 h for 3.853 and 1.859, respectively.
The main mechanism of the communication between EVs derived from HCF and their host remains unclear. Our results suggest that HC may modulate the expression of miRNAs, involved in liver fibrosis via EVs.
包虫病是一种由细粒棘球绦虫幼虫阶段引起的人畜共患被忽视疾病。有证据表明包虫囊肿（HC）与宿主之间通过细胞外囊泡（EVs）进行交流。然而，对于源自HC液（HCF）的EVs与宿主细胞之间的交流了解甚少。本研究旨在探讨HCF衍生的EVs对THP-1细胞系中纤维化和抗纤维化微小RNA（miRNAs）表达的影响。
在本研究中，通过超速离心从野生感染绵羊的HCF中分离出EVs，并通过蛋白质免疫印迹、电子显微镜和大小分布分析进行表征。在孵育3小时和24小时后，使用定量实时聚合酶链反应（qRT-PCR）研究EVs对参与肝纤维化的微小RNA（mir-16、mir-29a和mir-155）表达水平的影响。
蛋白质免疫印迹分析证实了CD63标志物的表达，而EVs样品中不存在钙连接蛋白和CD81。扫描电子显微镜和形态学显示为圆形囊泡。动态光散射分析显示平均大小分布直径为130.6纳米。mir-16和mir-29a的表达水平在3小时后分别显著上调至8.66和3.420，而在24小时后分别显著下调至3.853和1.859。
源自HCF 的EVs与其宿主之间交流的主要机制仍不清楚。我们的结果表明，HC可能通过EVs调节参与肝纤维化的miRNAs的表达。
