Effect Of PAS-Na On NF-κβ/NLRP3/Caspase-1 Pathway In Manganese-induced BV2 Cells

Objective Investigation the effects of PAS-Na on NF-κB/NLRP3/Caspase-1pathway in manganese（Mn）-induced BV2 Cells.Methods:The effect of different concentrations of Mn,PAS-Na,and N-acetylcysteine（NAC）on the viability of BV2 cells was examined using a cytotoxicity assay（MTT）,and the appropriate Mn concentration（200μmol/L）was selected.PAS-Na（100,200,400μmol/L）and NAC（50μmol/L）were the intervention doses.The following groups were identified:control group,PAS-Na control group,NAC control group,positive control group,Mn-exposed group,PAS-Na（100,200,and 400μmol/L）intervention group,NAC（50μmol）/L)intervention group,normal medium cultured for 24 hours.Then remove the original broth.Control group,PAS-Na control group and NAC control group were cultured with new medium culture,containing 400μmol/L PAS-Na medium and 50μmol/L NAC medium,respectively.The cells of manganese-exposured group and PAS-Na（100,200,and400μmol/L）intervention group were maintained in medium with 200μmol/L MnCl₂ for 24h.After removing the original culture solution,the control group,PAS-Na control group,NAC control group and manganese-exposured group were cultured fresh medium,and the PAS-Na intervention groups were respectively added fresh medium with 100,200 and 400μmol/L PAS-Na,cultured for 12,24,and 48 h.ROS production levels,mitochondrial membrane potential changes,and ATP production levels were determined in each group using the active oxygen test kit,mitochondrial membrane potential assay kit（JC-1）,ATP content assay kit.Expression of P65,NLRP3,caspsase-1,IL-1β,IL-18 mRNA were detected by q PCR.Results:Treatment with 50umol/L manganese increased the viability of BV2 cells.However,with the increase of manganese concentration,the survival rate of BV2 cells decreased,the difference was statistically significant（P<0.05）,and has a fixed time/dose-response relationship.PAS-Na can increase the survival rate of BV2 cells after intervention for 48 hours.With the PAS-Na concentration increasing,the survival rate gradually increased higher（P<0.05）.After Mn（200μmol/L）for 24h,the production of ROS in BV2 cells was increased.PAS-Na could antagonize the production of ROS induced by manganese.With the increase of the concentration and the prolongation of time,the production of ROS was lower（P<0.05）,but its intervention was no better than NAC（P<0.05）.After24 hours of manganese exposure,the mitochondrial membrane potential（MMP）and ATP production of BV2 cells were decreased（P<0.05）.PAS-Na intervention for 12 h had no obvious effect on improving mitochondrial membrane potential and ATP production（P>0.05）.But the effect was significant when the intervention time prolonged to 48h（P<0.05）.Manganese promoted the expression of NF-κB P65,NLRP3,caspase-1,IL-1βand IL-18 mRNA.PAS-Na could antagonistic the expression of P65,NLRP3,caspase-1 and IL-1βinduced by manganese（P<0.05）.However,IL-18 did not decline.Conclusion:（1）Excessive manganese exposure（200μmol/L）can induce oxidative damage,mitochondrial dysfunction,and ATP depletion in BV2 microglial cells.（2）Excess Mn exposure（200μmol/L）could induce inflammatory response in BV2 cells by mediating NF-κB/NLRP3/Caspase-1 inflammatory pathways.（3）PAS-Na can antagonize the oxidative damage and inflammation of BV2 cells induced by manganese.