Applied Physiology Laboratory, University of North Texas, Denton, TX, USA; Department of Biological Sciences, University of North Texas, Denton, TX, USA.
Applied Physiology Laboratory, University of North Texas, Denton, TX, USA.
Methods. 2019 Apr 1;158:77-80. doi: 10.1016/j.ymeth.2018.11.013. Epub 2018 Nov 22.
Bead-based analysis methods allow for the exploration of a variety of complex biological processes. In particular, these techniques can be applied to better understand how peripheral muscle injury contributes to systemic inflammation. Understanding how these two processes affect one another can give additional insight concerning how changes in inflammation effect readiness to perform in exercise and work environments. The present method sought to combine the strengths of bead-based multiplexing with the precision and low-end detection of single molecule counting (SMC) methods. We used performance of an extreme aerobic exercise session (i.e. half-marathon race) to cause a defined quantity of lower body muscle injury and a systemic inflammatory response lasting up to 24 h. Using a high-sensitivity, multiplex assay (Milliplex; Millipore-Sigma) we were able to identify 9 of 21 cytokines that were significantly elevated at either 4 or 24 h post half-marathon performance. Despite the known role of IL-1β, IL-6, and TNF-α in the pro-inflammatory response, they did not appear to change based on the multiplex analysis. We thus, conducted further analysis using an SMC assay and found increases in IL-1β, IL-6, and TNF-α at 4 h compared to 24 h post exercise. This method approach demonstrates how combining two common, bead-based protein assays can increase the amount of meaningful biological information that can be collected. We anticipate that this approach will be useful in a variety of inflammation-associated disease states.
基于珠体的分析方法可用于探索多种复杂的生物过程。特别是，这些技术可用于更好地理解外周肌肉损伤如何导致全身炎症。了解这两个过程如何相互影响，可以进一步了解炎症变化如何影响在运动和工作环境中的表现能力。本方法旨在结合珠体多重分析的优势与单分子计数（SMC）方法的精确性和低检测限。我们使用极端有氧运动（即半程马拉松比赛）来造成明确数量的下肢肌肉损伤和持续长达 24 小时的全身炎症反应。使用高灵敏度、多重分析（Milliplex；Millipore-Sigma），我们能够在半程马拉松比赛后 4 或 24 小时确定 21 种细胞因子中的 9 种显著升高。尽管已知白细胞介素-1β、白细胞介素-6 和肿瘤坏死因子-α在促炎反应中起作用，但它们似乎并未根据多重分析而改变。因此，我们使用 SMC 分析进行了进一步分析，发现与运动后 24 小时相比，IL-1β、IL-6 和 TNF-α在 4 小时时增加。这种方法结合了两种常见的基于珠体的蛋白质分析方法，可以增加可以收集的有意义的生物学信息的数量。我们预计这种方法将在各种与炎症相关的疾病状态中有用。
