Neuroinflammation Mediated By Activation Of Microglia NLRP3 Pathway By Serum Amyloid A

Objective This experiment intends to study the related mechanism of SAA activating the inflammatory pathway of microglia and mediating the apoptosis of HT22 neurons.Materials and methodsFirst,SAA was added to act on BV-2 and HT22 cells for 24 h,and the CCK8 method was used to observe whether it had cytotoxic effect.HT22 cells were then treated with SAAstimulated BV-2 culture medium for 24 h,and the CCK8 method was used to count the survival rate of HT22 again.Flow cytometry was used to determine the type of injury after detecting the decrease in the survival rate of HT22 cells.BV-2 cells were stimulated with SAA and then sequenced by m RNA transcriptomics to screen inflammation-related and other differentially expressed genes,and select the significantly up-regulated IL-1β.ResultsThe CCK8 assay showed that SAA itself had no direct toxic effect on BV-2 and HT22 cells.After co culture of BV-2 cells and HT22 cells with SAA,the survival rate of HT22 cells was decreased by CCK8 test,and the apoptosis of HT22 cells was detected by flow cytometry.m RNA transcriptome sequencing showed that SAA promoted the up-regulation of a series of inflammatory factors and cytoskeleton-related protein genes such as actin filaments,among which IL-1β,Acta1 and other genes were significantly differentially expressed.Western Blot showed that SAA activates BV-2 cells through the NLRP3 signaling pathway to increase the expression of IL-1β.ConclusionsSAA can stimulate the NLRP3 signaling pathway of microglia to release the inflammatory factor IL-1β and mediate neuronal apoptosis.