〔Abstract〕 Objective To investigate the effect of high concentration of Caerulomycin A(Cae A) on HK2 in renal tubular epithelial cells and to explore the role of cytoplasmic nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3) in this process. Methods The effect of different concentrations of Cae A on the viability of HK2 cells was determined by MTT; the expression of kidney injury molecule(KIM-1) and NLRP3 was detected by real-time quantitative PCR, Western blot and immunofluorescence, while the effect of Cae A on the mRNA expression of IL-1β, IL-18, IL-33, MCP-1, TNF-α was also measured by real-time quantitative PCR. HK2 cells were divided into control group, high concentration of Cae A group and high concentration of Cae A plus NLRP3 inhibitor CY-09 group, and the expression of KIM-1 and NLRP3 protein was detected by Western blot. Results The results of MTT showed that high concentration of Cae A could inhibit HK2 cell viability. Real-time quantitative PCR, Western blot and immunofluorescence assays showed that high concentration of Cae A upregulated the expression of KIM-1 and NLRP3, as well as the mRNA levels of IL-1β, IL-18, IL-33, MCP-1, TNF-α, while CY-09 could down-regulate the expression of NLRP3 and KIM-1. Conclusion High concentration of Cae A significantly inhibited the viability of HK2 cells and induced damage and inflammatory response to HK2 with some nephrotoxicity that might be achievedviaNLRP3 pathway.