Mancini Sarah J, Salt Ian P
Institute of Cardiovascular & Medical Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK.
Methods Mol Biol. 2018;1732:307-319. doi: 10.1007/978-1-4939-7598-3_20.
In addition to the well-characterized role of AMPK in the regulation of nutrient metabolism, it is increasingly clear that AMPK activation has multiple actions on inflammatory signalling. Here we describe methods to identify effects of AMPK activity on pro-inflammatory signalling, specifically (1) the nuclear localization of the key inflammatory mediators nuclear factor-κB (NFκB) and phosphorylated c-Jun N-terminal kinase (JNK), (2) preparation of conditioned medium to analyze the secretion of cytokines/chemokines, and (3) the pro-inflammatory adhesion of leukocytes to cultured cells.
除了AMPK在调节营养代谢方面已明确的作用外,越来越清楚的是,AMPK激活对炎症信号传导有多种作用。在这里,我们描述了鉴定AMPK活性对促炎信号传导影响的方法,具体包括:(1)关键炎症介质核因子-κB(NFκB)和磷酸化c-Jun氨基末端激酶(JNK)的核定位;(2)制备条件培养基以分析细胞因子/趋化因子的分泌;(3)白细胞与培养细胞的促炎黏附。