涤痰清脑方通过调节NF-κB/NLRP3通路减轻LPS诱导小胶质细胞神经炎症损伤OA

Objective To investigate the inhibitory effect of the Ditan Qingnao formula(DTS)on lipopolysaccharide(LPS)-induced neuroinflammation and to explore its molecular mechanism.Methods Microglia(BV2)were used as the experimental subjects,and were divided into blank group,model group(1 mg/L LPS intervention),and traditional Chinese medicine group(given three doses of 0.58 mg/mL,0.93 mg/mL,1.16 mg/mL DTS+1 mg/L LPS intervention,divided into three groups).Western blot was used to detect the levels of key proteins in the nuclear factor(NF)-κB/nucleotide-binding oligomerization domain-like receptor protein(NLRP)3 signaling pathway.ELISA was used to detect the expression of interleukin(IL)-1β/IL-18 and its precursors pro-IL-1β/pro-IL-18.Immunofluorescence assay detects microglial activation.Results Compared with the blank group,the NF-κB/NLRP3 signaling pathway in the model group was significantly activated,the levels of IL-18 and IL-1β were increased,the fluorescence intensity of DAPI,immune cell surface antigen(CD16),and ionic calcium-binding linker molecule-1(IBA-1)in BV2 microglia was increased,and the key proteins of the NF-κB/NLRP3 signaling pathway:The gray values of NF-κB p65 subunit,NLRP3 and Caspase-1 were significantly increased(P<0.05).Compared with the model group,the expression levels of IL-1β and IL-18,the grayscale value of NLRP3 and the grayscale value of Caspase-1 in the three dose groups of LPS+DTS were significantly decreased(P<0.05).The gray value of NF-κB p65 was significantly reduced in the medium dose group(P<0.05).The fluorescence intensity of CD16 and IBA-1 was significantly weakened in the medium and high dose groups(P<0.01).Conclusion The Ditan Qingnao formula can reduce LPS-induced neuroinflammation injury in BV2 microglial cells.The mechanism may be related to the formula's components reducing the activation level of the NF-κB/NLRP3 signaling pathway,thereby decreasing the expression of inflammatory factors and inhibiting neuroinflammation.