Inhibition of naringenin on lipopolysaccharide-induced pyroptosis of chondrocytes and its mechanism
Objective To observe the inhibitory effect of naringenin(NAR)on lipopolysaccharide(LPS)-induced pyroptosis of chondrocytes and to explore its possible mechanism.Methods C28/I2 cells in the logarithmic growth phase were divided into the LPS group(cultured with DMEM medium containing 100 ng/mL LPS),the LPS+NAR group(cultured with medium containing 100 ng/mL LPS+100 μmol/L NAR),the LPS+NAR+DORS group(cultured with medium containing 100 ng/mL LPS+100 μmol/L NAR+50 μmol/L DORS),and the Control group(cultured with ordi-nary medium),and all groups were cultured for 8 h.Cells in each group were taken.The cell proliferation optical density(OD)value was detected by the CCK-8 method,the apoptosis rate was detected by the TUNEL method,the relative pro-tein expression levels of pyroptosis markers[interleukin(IL)-18,GSDMD],adenosine-monophosphate-activated protein kinase α(AMPKα),phosphorylated AMPKα(p-AMPKα),NLRP3 inflammasome marker proteins[apoptosis-associated speck-like protein containing a CARD(ASC),Caspase-1),nucleotide-binding oligomerization domain-like receptor fami-ly pyrin domain-containing 3(NLRP3),and cleaved-Caspase-1]were detected by Western blotting,and the levels of IL-1β,IL-6,and tumor necrosis factor-α(TNF-α)in the cell supernatant were detected by ELISA.Results In the Control group,LPS+NAR group,LPS+NAR+DORS group,and LPS group,the cell proliferation OD values decreased succes-sively,the apoptosis rates increased successively,the relative protein expression levels of IL-18,GSDMD,ASC,Caspase-1,NLRP3,and cleaved-Caspase-1 in the cells increased successively,the relative protein expression level of p-AMPKα decreased successively,the levels of IL-1β,IL-6,and TNF-α in the cell supernatant increased successively while the lev-els of IL-10 and IL-13 decreased successively,with statistically significant difference between groups(all P<0.05).The relative protein expression levels of AMPKα in the LPS group and LPS+NAR+DORS group were both lower than those in the Control group and LPS+NAR group(all P<0.05).Conclusion NAR can inhibit LPS-induced chondrocyte pyropto-sis,and its mechanism may be related to activating the AMPKα signaling pathway and inhibiting the NLRP3 inflamma-some-mediated inflammatory response.
naringeninAMPKα signal pathwayNLRP3 inflammasomechondrocytepyroptosis
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