Le Maître Erwan, Revathikumar Priya, Estelius Johanna, Lampa Jon
Department of Medicine, Unit of Rheumatology, Center for Molecular Medicine (CMM), Karolinska Institute, Karolinska University Hospital;
Department of Medicine, Unit of Rheumatology, Center for Molecular Medicine (CMM), Karolinska Institute, Karolinska University Hospital.
J Vis Exp. 2017 Mar 29(121):54890. doi: 10.3791/54890.
Inflammation is a local response to infection and tissue damage mediated by activated macrophages, monocytes, and other immune cells that release cytokines and other mediators of inflammation. For a long time, humoral and cellular mechanisms have been studied for their role in regulating the immune response, but recent advances in the field of immunology and neuroscience have also unraveled specific neural mechanisms with interesting therapeutic potential. The so-called cholinergic anti-inflammatory pathway (CAP) has been described to control innate immune responses and inflammation in a very potent manner. In the early 2000s, Tracey and collaborators developed a technique that stimulates the vagus nerve and mimics the effect of the pathway. The methodology is based on the electrical stimulation of the vagus nerve at low voltage and frequency, in order to avoid any side effects of overstimulation, such as deregulation of heart rate variability. Electrical devices for stimulation are now available, making it easy to set up the methodology in the laboratory. The goal of this research was to investigate the potential involvement of prostaglandins in the CAP. Unfortunately, based on earlier attempts, we failed to use the original protocol, as the induced inflammatory response either was too high or was not suitable for enzymatic metabolism properties. The different settings of the original surgery protocol remained mostly unchanged, but the conditions regarding inflammatory induction and the time point before sacrifice were improved to fit our purposes (i.e., to investigate the involvement of the CAP in more limited inflammatory responses). The modified version of the original protocol, presented here, includes a longer time range between vagus nerve stimulation and analysis, which is associated with a lower induction of inflammatory responses. Additionally, while decreasing the level of lipopolysaccharides (LPS) to inject, we also came across new observations regarding mechanistic properties in the spleen.
炎症是由活化的巨噬细胞、单核细胞和其他免疫细胞介导的对感染和组织损伤的局部反应，这些细胞会释放细胞因子和其他炎症介质。长期以来，体液和细胞机制在调节免疫反应中的作用一直受到研究，但免疫学和神经科学领域的最新进展也揭示了具有有趣治疗潜力的特定神经机制。所谓的胆碱能抗炎途径（CAP）已被描述为以一种非常有效的方式控制先天免疫反应和炎症。在21世纪初，特雷西及其合作者开发了一种刺激迷走神经并模拟该途径作用的技术。该方法基于以低电压和频率对迷走神经进行电刺激，以避免过度刺激的任何副作用，如心率变异性失调。现在有用于刺激的电子设备，使得在实验室中建立该方法很容易。本研究的目的是调查前列腺素在CAP中的潜在作用。不幸的是，基于早期的尝试，我们未能使用原始方案，因为诱导的炎症反应要么过高，要么不适合酶代谢特性。原始手术方案的不同设置大多保持不变，但炎症诱导条件和处死前的时间点得到了改进以符合我们的目的（即研究CAP在更有限的炎症反应中的作用）。这里介绍的原始方案的修改版本包括迷走神经刺激和分析之间更长的时间范围，这与较低的炎症反应诱导相关。此外，在降低脂多糖（LPS）注射水平的同时，我们还发现了关于脾脏机制特性的新观察结果。
