Study On The Mechanism Of NF-κB-IL-6,IL-8,TNF-α Signaling Pathway In The Inflammatory Response Of Neurobrucellosis

Objective:Microglia models infected with Brucella glabrata 2308 were constructed to detect the transcription and the expression of IL-6,IL-8 and TNF-α.The assay was aimed to explore the relationship between NF-κB signaling pathway and expression of inflammatory factors.The effect of activating NF-κB signaling pathway might lay a foundation for the study of the inflammatory mechanism of neurobrucellosis;The expression of NF-κB in microglia was interfered.The effect of NF-κB on the expression of IL-6,IL-8 and TNF-α in the inflammatory response of neurobrucellosis could provide a new view for the anti-inflammatory treatment of neurobrucellosis.Methods: Human microglia HMC3 was infected with Brucella glabrata 2308.The transcription and expression of IL-6,IL-8 and TNF-α were detected by ELISA and PCR.The expression of NF-κB p65,IKK,p-p65,p-IKK were examined by Western blot.The microglia model infected with specific adenovirus vector to be used to show the over expression of NF-κB;with the over expression microglia model,siRNA interference was constructed to inhibit the expression of NF-κ B;the two cell models were infected with Brucella.The transcription and expression of IL-6,IL-8 and TNF-α were detected by ELISA and PCR.The infection group was compared with the control group.SPSS 22.0 software was used for statistical analysis.Results:(1)compared with the control group without Brucella infection,the expressions and levels of IL-6,IL-8 and TNF-α were significantly increased in the infected group at 8h(P < 0.05),the level of TNF-α was significantly increased at24 h after infection(P < 0.05).The expression of IL-6,IL-8 and TNF-α increased with the time of infection.(2)Compared with the blank control group,the expression of IL-6 mRNA,IL-8 mRNA and TNF-α mRNA in the infected group elevated.the increase of IL-8 mRNA at 4 h after infection(P < 0.05);the increase of IL-6 mRNA and TNF-α mRNA at 24 h after infection(P < 0.05).(3)the expression levels of p-p65 and p-IKK in 48 h infection group were significantly higher than the control(P< 0.05).(4)The expression levels of IL-6,IL-8 and TNF-α in adenovirus vector group were significantly higher than those in normal cell infection group(P < 0.05).The expression of IL-8 and TNF-α in siRNA group were significantly higher than that in blank group(P < 0.05).(5)The expression of IL-6 mRNA,IL-8 mRNA and TNF-α mRNA in adenovirus vector group was significantly higher than that in normal cell infection group(P < 0.05).The expression of IL-6 mRNA,IL-8 mRNA and TNF-α mRNA in siRNA group was significantly higher than that in normal cell infection group(P < 0.05).Conclusion:(1)NF-κB pathway was activated after Brucella infection,the expression of inflammatory factors IL-6,IL-8 and TNF-αwere increased,and the expressions were related to the infective duration,which may be due to the inflammatory loop formed by the continuous activation of NF-κB pathway.NF-κ B signaling pathway and inflammatory pathway of-Il-6,IL-8 and TNF-α might play important roles in the pathogenesis of neurobrucellosis.(2)NF-κB may have both pro-inflammatory and anti-inflammatory effects in the pathogenesis of neurobrucellosis.Inhibition of NF-κB may aggravate the inflammatory reaction of neurobrucellosis.The role of NF-κB in Brucella induced inflammation may be bidirectional.