| Objective:To study the m RNA relative expression levels of NOD-like receptor protein 1(NOD1),NOD-like receptor protein 2(NOD2)and nuclear factor-κB(NF-κB)and the expression levels of inflammatory cytokines including interleukin-6(IL-6)and interferon-γ(IFN-γ)in peripheral venous blood of acute and chronic brucellosis patients.The correlation between NOD1,NOD2,NF-κB and IL-6 and IFN-γwas analyzed.The occurrence and participation of these factors in the acute and chronic stages of brucellosis were discussed,and the possible mechanism of action was explored to provide possible clues for the study of the pathogenesis and chronic mechanism of brucellosis.Methods:A total of 50 patients with brucellosis admitted to the Center for Disease Control and Prevention of Inner Mongolia Autonomous Region from July 2022 to February 2023 were included in the case group(confirmed according to the national diagnostic criteria WS269-2019 for brucellosis),including 30 patients in the acute stage and 20 patients in the chronic stage.During the same period,20 healthy patients from the Physical Examination Center of Affiliated Hospital of Inner Mongolia Medical University were collected and included into the control group.All the subjects signed informed consent,collected age、gender and other basic information into the database,and collected 1ml fasting elbow venous blood in the morning.QRT-PCR were performed to determine the expression of NOD1,NOD2 and NF-κB in all samples.1ml of serum was obtained and the expression levels of IL-6 and IFN-γwere determined by ELISA.Statistical software SPSS 25.0 was used to process all data.The correlation of receptor proteins NOD1,NOD2,NF-κB and the inflammatory cytokines IL-6 and IFN-γwas analyzed.Results:The relative expression level of NOD1 receptor gene was 0.71(0.55,0.92)in patients with acute phase,0.53(0.36,0.76)in patients with chronic phase,and 0.39(0.31,0.68)in the control group.The overall differences of relative expression level of the three groups were statistically significant(χ~2=12.757,(49)=0.002),the differences between the acute stage group and control group(Z=3.264,(49)=0.001),acute and chronic phase(Z=2.634,(49)=0.008)were statistically significant((49)<0.05).There was no significant difference between chronic phase and the control group(Z=0.575,(49)>0.05).The relative expression level of NOD2 receptor gene was 0.66(0.47,0.81)in patients with acute stage,0.46(0.39,0.59)in patients with chronic stage and 0.39(0.30,0.61)in the control group.The overall differences of relative expression level of the three groups were statistically significant(χ~2=8.997,(49)=0.011),there was a statistically significant difference between the acute stage group and control group(Z=2.938,(49)=0.003),but no statistically significant difference was found between acute and chronic phase(Z=1.729,(49)=0.840),chronic phase and control group(Z=1.103,(49)=0.270)((49)>0.05).The relative expression level of NF-κB gene was 0.81(0.60,0.94)in acute phase,0.51(0.32,0.62)in chronic phase and 0.47(0.33,0.64)in the control group.The overall differences of relative expression level of the three groups were statistically significant(χ~2=18.930,(49)<0.001),the differences between the acute stage group and control group(Z=3.688,(49)<0.001),acute and chronic phase(Z=3.591,(49)<0.001)were statistically significant((49)<0.05).There was no significant difference between chronic phase and the control group(Z=0.089,(49)>0.05).The expression level of IL-6 was 52.60(48.17,55.90)in acute phase,65.05(45.56,74.00)in chronic phase,and 36.81(33.14,41.15)in the control group.The overall differences of IL-6 expression level among the three groups were statistically significant(χ~2=38.915,(49)<0.001),the differences between the acute phase and the control group(Z=5.260,(49)<0.001),chronic phase and the control group(Z=5.687,(49)<0.001)were statistically significant((49)<0.05).There was no significant difference between acute and chronic phase(Z=0.970,(49)>0.05).The expression level of IFN-γwas 525.29±71.94 in acute stage,464.36±64.70 in chronic stage,and 373.17±73.85 in the control group.The overall differences of IFN-γexpression level among the three groups was statistically significant(F=27.914,(49)<0.001).The differences between the acute stage group and the control group(F=7.248,(49)<0.001),acute and chronic phase(F=3.052,(49)=0.004),chronic phase and the control group(F=4.154,(49)<0.001)were statistically significant((49)<0.05).The correlation between NOD1,NOD2,NF-κB and IL-6,IFN-γwere analyzed with spearman rank correlation analysis,and there was a weak positive correlation between the expression level of NOD1 gene and IL-6 expression level(r=0.251,(49)=0.036).The relative expression of NF-κB gene was weakly positively correlated with IFN-γ(r=0.288,(49)=0.016).Conclusion:(1)NOD1 and NOD2 receptor genes showed significantly high levels of expression in acute stage patients with brucellosis,which may be activated in the early stage of human brucellosis infection and participate in the innate immune response;(2)The relative expression level of NF-κB and cytokines IL-6 and IFN-γincreased significantly in acute patients,and the relative expression level of NOD1 was weakly positively correlated with the expression level of IL-6.It is possible that by activating NF-κB to promote the production of inflammatory mediators such as IL-6 and IFN-γ,it mediates the innate immune inflammatory response induced by the up-regulated expression of NOD1 and NOD2.(3)The relative expression level of NF-κB and IFN-γincreased in acute patients,showing a weak positive correlation,suggesting that the two may have synergistic or mutually reinforcing effects in the early stage of brucellosis,or there may be some factors related to the activation of them.(4)NOD1 and NOD2 still showed a low level of elevated expression in the chronic stage,which may be due to the gradual dominance of some factors enhanced in the later stage of infection,rather than being completely replaced or eliminated.This effect may be partly involved in the continued high expression of IL-6 in the chronic stage. |
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