α-mangostin inhibits LPS/ATP-induced NLRP3 inflammasome activation in microglia via the NF- κB pathway / 安徽医科大学学报

ABSTRACT

Objective @#To investigate the effects and underlying mechanisms of α-mangostin in a spinal cord inj ury 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 reduced (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) . C ompared 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, and the p-p65/p65 ratio in the 40 μmol/L α-mangostin group , as well as the expression 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 ob served 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 .