摘　要：NOD样受体热蛋白结构域相关蛋白3（NLRP3）炎性小体是一种蛋白复合体，在机体抵抗感染以及无菌性炎症中发挥了至关重要的作用。在HEK293T细胞中构建NLRP3炎性小体活化体系有助于研究其活化及调节机制。将质粒NLRP3、凋亡相关点样蛋白（ASC）、胱冬肽酶-1（Caspase-1）以及pro-1β转染至HEK293T细胞中，随后加入尼日利亚菌素（Nigericin），并利用Western blot以及酶联免疫吸附试验（ELISA）测定成熟的IL-1β。试验结果表明：转染了完整NLRP3炎性小体组分的细胞蛋白和上清中均可检测到IL-1β的成熟体，反之，未转染完整组分的细胞蛋白及上清中不能检测到IL-1β成熟体；同时，随着NLRP3转染量的增加，可检测到的IL-1β的成熟体增加。本研究在HEK293T细胞中成功构建了NLRP3炎性小体活化体系，为后续研究奠定了基础。
关键词：NLRP3炎性小体；HEK293T细胞；质粒转染；IL-1β成熟体；抗感染
中图分类号：R392                                 文献标志码：A                     DOI：10.3969/j.issn.1007-7146.2022.04.006

Abstract: NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome is a multiprotein complex that plays a crucial role in host immune responses to both pathogen infections and sterile injuries. Construction of the NLRP3 inflammasome system in HEK293T cells provides a more convenient and concise method for studying the regulation mechanisms of inflammasome activation. Plasmids NLRP3, apoptosis-associated speck-like protein (ASC), Caspase-1, and pro-1β were transfected into HEK293T cells. Subsequently, the mature IL-1β levels were determined by Western blot and enzyme-linked immunosorbent assay (ELISA) after adding nigericin. Our results showed that the mature IL-1β could be detected in cell lysates and supernatants of cells transfected with intact NLRP3 inflammasome components whereas not in cells transfected with incomplete NLRP3 inflammasome components. Meanwhile, the mature IL-1β increased with the elevated amounts of transfected NLRP3. In this study, NLRP3 inflammasome system was successfully constructed in HEK293T cells, which laid a foundation for future research.
Key words: NLRP3 inflammasome; HEK293T cells; plasmid transfection; mature IL-1β; anti infection
（Acta Laser Biology Sinica, 2022, 31(4): 331-336）