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从包虫囊液中分离的细胞外囊泡的表征及其对人单核细胞免疫调节作用的评价。

Characterisation of extracellular vesicles isolated from hydatid cyst fluid and evaluation of immunomodulatory effects on human monocytes.

机构信息

Department of Pharmacy and Pharmaceutical Technology and Parasitology, University of Valencia, Valencia, Spain.

Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

出版信息

J Cell Mol Med. 2023 Sep;27(17):2614-2625. doi: 10.1111/jcmm.17894. Epub 2023 Aug 2.

Abstract

Hydatidosis is a disease caused by the larval stage of Echinococcus granulosus, which involves several organs of intermediate hosts. Evidence suggests a communication between hydatid cyst (HC) and hosts via extracellular vesicles. However, a little is known about the communication between EVs derived from HC fluid (HCF) and host cells. In the current study, EVs were isolated using differential centrifugation from sheep HCF and characterized by western blot, electron microscope and size distribution analysis. The uptake of EVs by human monocyte cell line (THP-1) was evaluated. The effects of EVs on the expression levels of pro- and anti-inflammatory cytokines were investigated using quantitative real-time PCR (RT-PCR), 3 and 24 h after incubation. Moreover, the cytokine level of IL-10 was evaluated in supernatant of THP-1 cell line at 3 and 24 h. EVs were successfully isolated and showed spherical shape with size distribution at 130.6 nm. After 3 h, the expression levels of pro-inflammatory cytokine genes (IL1Β, IL15 and IL8) were upregulated, while after 24 h, the expression levels of pro-inflammatory cytokines were decreased and IL13 gene expression showed upregulation. A statistically significant increase was seen in the levels of IL-10 after 24 h. The main mechanism of the communication between EVs derived from HCF and their host remains unclear; however, time-dependent anti-inflammatory effects in our study suggest that HC may modulate the immune responses via EVs.

摘要

包虫病是由细粒棘球绦虫的幼虫阶段引起的疾病,涉及中间宿主的几个器官。有证据表明,包囊液(HCF)中的细胞外囊泡(EVs)与宿主之间存在通讯。然而,对于 HCF 来源的 EVs 与宿主细胞之间的通讯,我们知之甚少。在本研究中,使用差速离心法从绵羊 HCF 中分离 EVs,并通过 Western blot、电子显微镜和粒径分布分析对其进行表征。评估 EVs 被人单核细胞系(THP-1)摄取的情况。通过定量实时 PCR(RT-PCR)在孵育 3 和 24 小时后,研究 EVs 对促炎和抗炎细胞因子表达水平的影响。此外,在 3 和 24 小时时,评估了 THP-1 细胞系上清液中 IL-10 的细胞因子水平。成功分离出 EVs,其形状呈球形,粒径分布在 130.6nm。孵育 3 小时后,促炎细胞因子基因(IL1Β、IL15 和 IL8)的表达水平上调,而 24 小时后,促炎细胞因子的表达水平下降,IL13 基因表达上调。24 小时后,IL-10 水平明显升高。HCF 来源的 EVs 与其宿主之间的通讯的主要机制尚不清楚;然而,我们的研究中显示的时间依赖性抗炎作用表明,HC 可能通过 EVs 调节免疫反应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9372/10468670/713cdf5e0947/JCMM-27-2614-g004.jpg

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