Mo Yiqun, Zhang Yue, Zhang Qunwei
Department of Environmental and Occupational Health Sciences, School of Public Health and Information Sciences, University of Louisville, Louisville, KY, USA.
Methods Mol Biol. 2019;1894:313-322. doi: 10.1007/978-1-4939-8916-4_18.
Nanoparticles can enter the body via the lungs, the digestive tract, and the skin. The pulmonary effects of nanoparticles and the underlying mechanisms may be investigated by histopathological examination, immunostaining, bronchoalveolar lavage (BAL), and so on. BAL is a simple, fast, and inexpensive method that is commonly used to investigate the immune cells and acellular components in the lungs, which reflect the health state of the lungs. In this chapter, we describe the detailed procedures of performing BAL on mouse lungs. In a typical procedure, a catheter is inserted in the exposed trachea of a mouse, through which a phosphate-buffered saline (PBS) solution containing ethylenediaminetetraacetic acid (EDTA) is instilled into the lungs by a syringe. BAL fluid (BALF) is collected by gently retracting the instilled solution and massaging the thoracic cavity to maximize BALF retrieval and to minimize shearing forces. The type and number of differentiated immune cells in the BALF are analyzed, and the soluble factors such as total proteins, chemokines, and cytokines in the BALF are determined by protein assay, ELISA, etc.
纳米颗粒可通过肺部、消化道和皮肤进入人体。纳米颗粒的肺部效应及其潜在机制可通过组织病理学检查、免疫染色、支气管肺泡灌洗(BAL)等方法进行研究。BAL是一种简单、快速且廉价的方法,常用于研究肺中的免疫细胞和无细胞成分,这些成分反映了肺的健康状况。在本章中,我们描述了对小鼠肺部进行BAL的详细步骤。在典型的操作过程中,将一根导管插入小鼠暴露的气管中,通过注射器经此导管将含有乙二胺四乙酸(EDTA)的磷酸盐缓冲盐水(PBS)溶液注入肺中。通过轻轻回抽注入的溶液并按摩胸腔来收集支气管肺泡灌洗液(BALF),以最大限度地回收BALF并最小化剪切力。分析BALF中分化免疫细胞的类型和数量,并通过蛋白质测定、酶联免疫吸附测定(ELISA)等方法测定BALF中的可溶性因子,如总蛋白、趋化因子和细胞因子。