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. 2021 Jul 16;14:3393–3403. doi: 10.2147/JIR.S311924

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© 2021 Li et al.

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Overexpression of NLRP3 reversed the suppressive effects of quercetin on IL-1β-induced rat chondrocyte injuries in vitro. Chondrocytes in the IL-1β + quercetin group were transiently transfected with an NLRP3 overexpression plasmid or empty vector. (A) Chondrocyte viability was analyzed by the CCK-8 assay. (B) The levels of pro-inflammatory cytokines (IL-18 and TNF-α) were measured by ELISA. (C) Cell apoptosis was evaluated by flow cytometry performed with an Annexin V-FITC/propidium iodide (PI) apoptosis kit. (D) Hoechst 33342 staining was performed. Apoptotic cells exhibited morphological changes in the nuclei typical of apoptosis. Images were captured under a fluorescence microscope. (E) The intracellular levels of ROS in chondrocytes were assessed using a DCFH-DA probe. (F) The levels of NLRP3, ASC, IRAK1, iNOS, COX-2 and caspase 3 proteins were assessed by Western blotting. Data represent a mean value ± standard deviation. **p < 0.01, ***p < 0.001, compared with blank; ^#p < 0.05, ^##p < 0.01, compared with IL-1β; ^$p < 0.05, ^$$p < 0.01, compared with IL-1β + quercetin + vector.