Cardiff School of Biosciences, Cardiff University, Cardiff, UK.
Methods Mol Biol. 2022;2419:313-331. doi: 10.1007/978-1-0716-1924-7_20.
Atherosclerosis is driven by chronic inflammation in all stages of the disease. Inflammation is fueled by elevated levels of pro-inflammatory cytokines. Interleukins (IL) are cytokines of particular importance in atherosclerosis, due to their key involvement in various pro-atherogenic processes, including infiltration of immune cells to the lesion, stimulation of the production of other pro-inflammatory mediators by other sources, and generation of lipid laden foam cells, all of which contribute to plaque development and progression. Various stimuli that are abundant in atherosclerotic plaques, including oxidized low-density lipoprotein, cholesterol crystals and reactive oxygen species can trigger inflammasome activation. Importantly, activation of the nucleotide oligomerization domain leucine-rich repeat and pyrin domain containing protein 3 (NLRP3) inflammasome activates the caspase-1 protease and results in the generation and release of potent pro-inflammatory cytokines, IL-1β and IL-18. Both cytokines are influential in driving chronic inflammation and atherogenesis. This chapter describes the use of enzyme-linked immunosorbent assay (ELISA) and Western blot to quantify these cytokines in cell supernatant and lysate respectively, after stimulating inflammasome activation in cultured cells.
动脉粥样硬化是由疾病各阶段的慢性炎症驱动的。炎症是由促炎细胞因子水平升高引起的。白细胞介素 (IL) 在动脉粥样硬化中尤为重要，因为它们在各种促动脉粥样硬化过程中起着关键作用，包括免疫细胞浸润到病变部位、刺激其他来源产生其他促炎介质，以及产生富含脂质的泡沫细胞，所有这些都有助于斑块的形成和发展。动脉粥样硬化斑块中大量存在的各种刺激物，包括氧化型低密度脂蛋白、胆固醇晶体和活性氧，可触发炎性小体的激活。重要的是，核苷酸寡聚化结构域富含亮氨酸重复序列和富含吡喃结构域蛋白 3 (NLRP3) 炎性小体的激活会激活半胱天冬酶-1 蛋白酶，导致强效促炎细胞因子白细胞介素-1β (IL-1β) 和白细胞介素-18 (IL-18) 的产生和释放。这两种细胞因子都在驱动慢性炎症和动脉粥样硬化形成中发挥重要作用。本章描述了在培养细胞中刺激炎性小体激活后，分别使用酶联免疫吸附测定 (ELISA) 和 Western blot 来定量细胞上清液和裂解物中的这些细胞因子。
