| Background:NLRP3 inflammasome(NLRP3)plays an important part in cerebral ischemia-reperfusion inflammatory injury.Activation of the NLRP3 inflammasome is closely related to the Toll-like receptor 4(TLR4)pathway.MJ33(1-hexadecyl-3-(trifluoroethgl)-sn-glycerol-2 phosphomethanol)is a special inhibitor of Acidic Calcium-independent phospholipase A2(aiPLA2)of Peroxiredoxin 6(Prdx6).MJ33 can decrease IL-1? and IL-18 in rats and may be related to TLR4.Objective:In this study,we used MJ33 to investigate whether it inhibits NLRP3 inflammasome and its possible mechanism.Methods:(1)Rats(n=72)were randomly divided into Sham group,MCAO group,NS group,MJ33 group,TLR4 inhibitor(TLR4 inh)group,DMSO group,MJ33 TLR4inhibitor(MT)group and solvent control(NS DMSO)group;(2)The model of middle cerebral artery occlusion in rats was established.The left common carotid artery was isolated in Sham group but not incised.Other groups were embolized for 1 hour and pulled out.MJ33 salt solution was injected into the tail vein of the MJ33 group before the operation.The same volume of saline was injected into the tail vein of NS group.DMSO solution was injected intraperitoneally in TLR4 inh group and equal volume DMSO was injected intraperitoneally in DMSO group after operation.In MT group,TLR4 inhibitor was injected intraperitoneally after the operation was performed by injecting MJ33,into the tail vein before the operation.In the solvent control group,normal saline and DMSO were injected in turn;(3)After 24 hours of cerebral ischemia reperfusion,the neurologic scores of Sham,MCAO,NS and MJ33 groups were evaluated;(4)The infarct size of Sham group,MCAO group,NS group and MJ33 group was detected by TTC assay;(5)WB was used to detect the activity of NLRP3 and related inflammatory factors,as well as the activity of TLR4 pathway-related proteins;(6)The expression of NLRP3 and the survival of neurons in each group were detected by immunofluorescence.Results:(1)The neurological function score and the range of cerebral infarction:Compared with the MCAO group,the neurological functional score and the range of cerebral infarction in the MJ33 group were significantly improved,and there was no statistical difference between the NS group and the MCAO group;(2)The results of immunofluorescence showed that the survival of neurons in MJ33 group was significantly better than that in MCAO group,while the expression of NLRP3 was significantly decreased;(3)The results of WB showed that NLRP3,IL-1 ?,IL-18 and Cleaved Caspase1 in MJ33 group were significantly lower than those in MCAO group,and TLR4 and MyD88 in MJ33 group were significantly lower than those in MCAO group;(4)The results of WB showed that TLR4 and MyD88 in TLR4 inh group were significantly inhibited(P=0.000 vs.MCAO);NLRP3,IL-1 ?,IL-18 and Cleaved Caspase1 in this group were significantly lower than those in MCAO group,but there was no significant difference between DMSO group and TLR4 inh group;(5)The results of immunofluorescence showed that the survival of neurons in TLR4 inh group was better than that in MCAO group,and the neuronal survival activity in MT group was further improved.The expression of NLRP3 in TLR4 inhgroup was significantly lower than that in MCAO group,while the activity of NLRP3 in MT group was further lower than that in TLR4 inh group.There was statistical significance in all groups,but there was no statistical difference between MCAO group and DMSO,Veh group.Conclusions:MJ33 plays a protective role in cerebral ischemia-reperfusion injury in rats,which is related to its anti-NLRP3 inflammatory body activation,which may be mediated by TLR4/MyD88 pathway. |
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