Department of Medicine, University of Geneva, 1206 Geneva, Switzerland.
Swiss Centre for Applied Human Toxicology, 4055 Basel, Switzerland.
Int J Mol Sci. 2021 May 11;22(10):5065. doi: 10.3390/ijms22105065.
Over the last decade, the knowledge in extracellular vesicles (EVs) biogenesis and modulation has increasingly grown. As their content reflects the physiological state of their donor cells, these "intercellular messengers" progressively became a potential source of biomarker reflecting the host cell state. However, little is known about EVs released from the human brain microvascular endothelial cells (HBMECs). The current study aimed to isolate and characterize EVs from HBMECs and to analyze their EVs proteome modulation after paraquat (PQ) stimulation, a widely used herbicide known for its neurotoxic effect. Size distribution, concentration and presence of well-known EV markers were assessed. Identification and quantification of PQ-exposed EV proteins was conducted by data-independent acquisition mass spectrometry (DIA-MS). Signature pathways of PQ-treated EVs were analyzed by gene ontology terms and pathway enrichment. Results highlighted that EVs exposed to PQ have modulated pathways, namely the ubiquinone metabolism and the transcription HIF-1 targets. These pathways may be potential molecular signatures of the PQ-induced toxicity carried by EVs that are reflecting their cell of origin by transporting with them irreversible functional changes.
在过去的十年中，细胞外囊泡（EVs）的生物发生和调节的知识不断增加。由于其内容反映了供体细胞的生理状态，这些“细胞间信使”逐渐成为反映宿主细胞状态的生物标志物的潜在来源。然而，人们对人脑血管内皮细胞（HBMEC）释放的 EVs 知之甚少。本研究旨在分离和鉴定 HBMECs 的 EVs，并分析其在百草枯（PQ）刺激后的 EVs 蛋白质组调节，百草枯是一种广泛使用的除草剂，具有神经毒性作用。评估了大小分布、浓度和已知 EV 标志物的存在。通过数据非依赖性采集质谱（DIA-MS）进行 PQ 暴露 EV 蛋白的鉴定和定量。通过基因本体术语和途径富集分析 PQ 处理的 EVs 的特征途径。结果表明，暴露于 PQ 的 EVs 调节了途径，即泛醌代谢和转录 HIF-1 靶标。这些途径可能是 EV 携带的 PQ 诱导毒性的潜在分子特征，通过携带不可逆的功能变化反映其起源细胞。
