Unidad de Oncogenómica, Servicio de Oncohematología, Fundación para la Investigación Biomédica del Hospital Universitario Niño Jesús, Madrid, Spain.
Methods Mol Biol. 2025;2876:61-75. doi: 10.1007/978-1-0716-4252-8_4.
The generation of hypothyroid and hyperthyroid mouse models is one of the approaches used to investigate the complex interplay between thyroid hormones and the immune system. We present a detailed protocol describing how to induce endotoxic shock by lipopolysaccharide (LPS) administration, and how to investigate the role of immune populations, specifically macrophages, responding to endotoxemia.This book chapter provides the use of different molecular techniques, such as Western Blotting, Immunohistochemistry, q-PCR, Luciferase assays, or ChIP assays, with which researchers can gain valuable insights into the immune system's interaction with hormonal signaling pathways, for instance, examining the effect of thyroid hormones on signaling of STAT3, NF-κB, and ERK in response to LPS, and inflammatory mediators, such as interleukin-6 (IL-6) or tumor necrosis factor-alpha (TNFα) within these cells. The signaling pathways involved and the exploration of the relationship between thyroid hormones and the immune system can be analyzed using several molecular biology technologies in order to clarify their interplay in various disease states.
制作甲状腺功能减退和甲状腺功能亢进的小鼠模型是研究甲状腺激素与免疫系统之间复杂相互作用的方法之一。我们提供了一个详细的方案,描述如何通过给予脂多糖(LPS)来诱导内毒素休克,以及如何研究免疫群体(特别是巨噬细胞)对内毒素血症的反应。本章介绍了使用不同的分子技术,如 Western Blotting、免疫组织化学、q-PCR、荧光素酶测定或 ChIP 测定等,研究人员可以通过这些技术深入了解免疫系统与激素信号通路的相互作用,例如,研究甲状腺激素对 STAT3、NF-κB 和 ERK 信号通路的影响,以及细胞内白细胞介素-6(IL-6)或肿瘤坏死因子-α(TNFα)等炎症介质的影响。可以使用几种分子生物学技术来分析涉及的信号通路,并探索甲状腺激素与免疫系统之间的关系,以阐明它们在各种疾病状态下的相互作用。