OBJECTIVE  To investigate the toxicity of genipin on human tubular epithelial cells(HK-2) based on NLRP3 pathway.

METHODS  The cytotoxic concentration and action time of genipin on HK-2 were detected by CCK8. The Western blotting was used to detect the expression of Kim-1, OPN and NLRP3 pathway protein NLRP3, Caspase-1, IL-1β and IL-18. Studied on the toxic effects of genipin on HK-2 cells, apoptosis in HK-2 cell was detected by Hoechst/PI staining, and the level of ROS in HK-2 cell was detected by DCFH-DA. High-content imaging techniques was used to detect mitochondrial membrane potential(MMP) and Ca²⁺ levels, Western blotting and qPCR methods was used to detect the expression levels of Kim-1, OPN, NLRP3, Caspase-1, IL-1β, IL-18 protein and mRNA. To study the antagonistic effect of NLRP3 inhibitor glyburide on GP toxicity and its mechanism.

RESULTS The IC₅₀ values of genipin for HK-2 cell were 433.00, 110.50 and 72.99 μg·mL^–1 at 12, 24 and 48 h, respectively. Compared with the blank group, after the treatment of genipin, the positive rate of PI, ROS, Ca²⁺ levels of HK-2 cell increased significantly, MMP decreased significantly, and the levels of Kim-1, OPN, IL-1β, IL-18, NLRP3, Caspase-1 protein and mRNA were significantly increased(P<0.05, P<0.01), and the NLRP3 pathway inhibitor glyburide could significantly improve the cytotoxic effects of genipin and related molecular changes.

CONCLUSION Genipin might have cytotoxicity on HK-2 cells via NLRP3 pathway, and inhibiting NLRP3 pathway can improve the inflammation caused by genipin induced cytotoxicity on HK-2 cells.