Study On The Protective Effect Of Sulforaphane On HMC3 Human Microglia Infected By Brucella Based On NF-κB Pathway

Objective:In this study,Sulforaphane(SFN)was used to interfere with human microglia infected with HMC3 by smooth strain B.abortus S2308,detecting the expression level of inflammatory cytokines tumor necrosis factor-α(TNF-α),interleukin-8(IL-8),interleukin-6(IL-6)and the transcription level of these inflammatory factors and nuclear factor-κB in microglia cells to explore the potential protective mechanism of Sulforaphane against brucella infection of microglia cells.Methods : Using a model of microglial cell injury induced by Brucella infection,human microglial cell HMC3 was randomly divided into four groups: blank control group(aseptic distilled water),infection group,SFN group 1(smooth strain B.abortus S2308+sulforaphane 10 umol/L group),and SFN group 2(smooth strain B.abortus S2308+sulforaphane 40 umol/L group).The blank control group was added with the same amount of aseptic distilled water as the drug group and the infection group was added with smooth strain B.abortus S2308 and the same amount of sterile physiological saline,and the drug group was added with smooth strain B.abortus S2308 and different concentrations of sulforaphane,respectively.After detecting cell viability using CCK8 method,ELISA and RT-q PCR were used to detect the expression of inflammatory factors TNF-α,IL-8,and IL-6and the transcription level of these inflammatory factors and NF-κB m RNA.Repeat each group of experiments three times.Statistical software SPSS 27.0 was used for analysis,and the measurement data were statistically described by mean differences ± standard deviation.t test was used for comparison between two groups,and one-way analysis of variance was used for comparison between groups.Bonferroni method was used to determine the statistical significance of differences between groups.The test level was set as α=0.05,and the statistical significance of differences was set as P<0.05.Results:Compared with blank group,the infection group had significant increases in the expression levels of inflammatory cytokines TNF-α,IL-8 and IL-6(P<0.05).Compared with infection group,the expression levels of inflammatory cytokines TNF-α,IL-8 and IL-6 in SFN group(sulforaphane 10umol/L and sulforaphane 40umol/L)were significantly decreased,with statistical significance(P<0.05).The inflammatory factor IL-6 in SFN group 2(sulforaphane 40umol/L)was significantly lower than that in SFN group 1(sulforaphane10umol/L),and there was statistical significance in their differences(P<0.05).Compared with blank group,the m RNA transcription of TNF-α,IL-8,IL-6 and NF-κB gene was up-regulated in infected group,and there was statistical significance in their differences(P<0.05).Compared with infection group,the m RNA transcription of TNF-α,IL-8,IL-6 and NF-κB gene in drug group(sulforaphane 10umol/L and sulforaphane 40umol/L)were decreased,with statistical significance(P<0.05).The m RNA transcription of inflammatory cytokines TNF-α,IL-8 and IL-6 in SFN group 2(sulforaphane 40umol/L)was significantly decreased compared with that in SFN group 1(sulforaphane 10umol/L),the difference was statistically significant(P<0.05),while the m RNA transcription of NF-κB was decreased,but there was not statistical significance in their differences(P>0.05).Conclusion:Sulforaphane can regulate the transcription of NF-κB and reduce the production of inflammatory cytokines TNF-α,IL-8 and IL-6 in microglia infected by brucella.Sulforaphane plays a neuroprotective role by regulating NF-κB/TNF-α,IL-8 and IL-6signaling pathways.