α-mangostin inhibits LPS/ATP-induced NLRP3 inflammasome activation in microglia via the NF-κB pathway
Objective To investigate the effects and underlying mechanisms of α-mangostin in a spinal cord injury model of microglial cell inflammation.Methods Mouse microglial cell line BV-2 was cultured in vitro,and an in-flammation model was established by co-treatment with lipopolysaccharide and adenosine triphosphate(LPS/ATP).The CCK-8 assay was used to test the influence of different concentrations(0,10,20,40,80 μmol/L)of α-man-gostin on cell proliferation vitality under LPS/ATP stimulation to select an appropriate concentration range of α-mangostin;BV-2 cells were divided into Ctrl group,LPS/ATP group,40 μmol/L α-mangostin group,and inter-vention groups with different concentrations(10,20,40 μmol/L)of α-mangostin(designated as LPS/ATP+10 μmol/L α-mangostin group,LPS/ATP+20 μmol/L α-mangostin group,and LPS/ATP+40 μmol/L α-mangostin group,respectively).ELISA experiments were conducted to detect the levels of pro-inflammatory cytokines inter-leukin-6/1β/18(IL-6,IL-1β,IL-18)and tumor necrosis factor(TNF-α)in the supernatants of each group,and Western blot was used to detect the expression of NLRP3,ASC,cleaved caspase-1,IL-1β,and the phosphoryla-tion levels of p65(p-p65/p65)in the NF-κB pathway,as well as the expression of p65 in the nuclei of BV-2 cells.Results Compared with the Ctrl group,cell proliferation vitality in the LPS/ATP group was significantly re-duced(P<0.05),but low concentrations(10,20,40 μmol/L)of α-mangostin significantly improved the inhibi-tory effect of LPS/ATP on microglial cell proliferation vitality(P<0.05),while a high concentration(80 μmol/L)of α-mangostin exacerbated the damage to microglial cells caused by LPS/ATP(P<0.05).Compared with the Ctrl group,the levels of inflammatory factors IL-6,IL-1β,IL-18,TNF-α,and the expression of NLRP3,ASC,cleaved caspase-1,IL-1β,andthep-p65/p65 ratiointhe40 μmol/Lα-mangostingroup,aswellastheexpression of p65 protein in the nuclei,showed no significant changes(P>0.05),whereas these significantly increased in the LPS/ATP group(P<0.05).Compared with the LPS/ATP group,the levels of IL-6,IL-1β,IL-18,TNF-α,and the expression of NLRP3,ASC,cleaved caspase-1,IL-1β,and the p-p65/p65 ratio in the intervention groups,as well as the expression of p65 protein in the nuclei,decreased in a concentration-dependent manner with increasing α-mangostin concentration,with the most significant reduction observed in the LPS/ATP+40 μmol/L α-mangostin group(P<0.01).Conclusion α-mangostin can inhibit the neuroinflammatory response mediated by NLRP3 inflammasome activation in BV-2 cells through the NF-κB pathway.
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