Pulliero A, Pergoli L, LA Maestra S, Micale R T, Camoirano A, Bollati V, Izzotti A, DE Flora S
Department of Health Sciences, University of Genoa, Italy.
EPIGET LAB, Department of Clinical Sciences and Community Health, University of Milan, Italy.
J Prev Med Hyg. 2019 Dec 20;60(4):E327-E336. doi: 10.15167/2421-4248/jpmh2019.60.4.1284. eCollection 2019 Dec.
Extracellular vesicles (EVs) are released from cells and enter into body fluids thereby providing a toxicological mechanism of cell-cell communication. The present study aimed at assessing (a) the presence of EVs in mouse body fluids under physiological conditions, (b) the effect of exposure of mice to cigarette smoke for 8 weeks, and (c) modulation of smoke-related alterations by the nonsteroidal anti-inflammatory drug celecoxib, a selective cyclooxygenase-2 inhibitor. To this purpose, ICR (CD-1) mice were either unexposed or exposed to cigarette smoke, either treated or untreated with oral celecoxib. EVs, isolated from bronchoalveolar lavage fluid (BALF), blood serum, and urines, were analyzed by nanoparticle tracking analysis and flow cytometry. EVs baseline concentrations in BALF were remarkably high. Larger EVs were detected in urines. Smoking increased EVs concentrations but only in BALF. Celecoxib remarkably increased EVs concentrations in the blood serum of both male and female smoking mice. The concentration of EVs positive for EpCAM, a mediator of cell-cell adhesion in epithelia playing a role in tumorigenesis, was much higher in urines than in BALF, and celecoxib significantly decreased their concentration. Thus, the effects of smoke on EVs concentrations were well detectable in the extracellular environment of the respiratory tract, where they could behave as delivery carriers to target cells. Celecoxib exerted both protective mechanisms in the urinary tract and adverse systemic effects of likely hepatotoxic origin in smoke-exposed mice. Detection of EVs in body fluids may provide an early diagnostic tool and an end-point exploitable for preventive medicine strategies.
细胞外囊泡（EVs）从细胞中释放出来并进入体液，从而提供了一种细胞间通讯的毒理学机制。本研究旨在评估：（a）生理条件下小鼠体液中EVs的存在情况；（b）小鼠暴露于香烟烟雾8周的影响；（c）非甾体抗炎药塞来昔布（一种选择性环氧化酶-2抑制剂）对烟雾相关改变的调节作用。为此，将ICR（CD-1）小鼠分为未暴露于香烟烟雾组和暴露于香烟烟雾组，每组再分为口服塞来昔布治疗组和未治疗组。通过纳米颗粒跟踪分析和流式细胞术对从支气管肺泡灌洗液（BALF）、血清和尿液中分离出的EVs进行分析。BALF中EVs的基线浓度非常高。在尿液中检测到较大的EVs。吸烟会增加EVs的浓度，但仅在BALF中。塞来昔布显著增加了雄性和雌性吸烟小鼠血清中EVs的浓度。上皮细胞中细胞间粘附介质EpCAM阳性的EVs浓度在尿液中比在BALF中高得多，且塞来昔布显著降低了其浓度。因此，在呼吸道的细胞外环境中可以很好地检测到烟雾对EVs浓度的影响，在该环境中它们可能作为靶向细胞的递送载体。塞来昔布在暴露于烟雾的小鼠中对尿路发挥了保护机制，但也产生了可能源于肝毒性的不良全身效应。检测体液中的EVs可能提供一种早期诊断工具以及可用于预防医学策略的终点指标。
