Expression And Significance Of TOLL Receptor 9 And Its Related Factors In Brucellosis Patients At Different Stages

Objective: Explore the immune mechanism of TLR9-My D88 signal pathway in human brucellosis,and clarify the dynamic changes of IL-4,IL-6,IL-12 and IFN-γ cytokines in brucellosis,providing evidence for the immune research of human brucellosis.Methods: From June 2022 to February 2023,50 cases of brucellosis were collected from the Inner Mongolia Center for Disease Control and Prevention,including 30 cases in the acute phase and 20 cases in the chronic phase.During the same period,in the Disease Physical Examination,collected 20 healthy people as the control grope of Inner Mongolia Medical University Hospital.Collected 1ml fasting elbow venous blood and measured m RNA expression levels of TLR9,My D88,NF-κb and IRF-7 in peripheral blood by q PCR.Serum1 ml was collected and ELISA kit was used to detect the expression levels of inflammatory cytokines IL-4,IL-6,IL-12 and IFN-γ.To analyze the correlation between TLR9 and My D88 signaling pathway and inflammatory factors,analyze the dynamic changes of cytokines in brucellosis patients,and explore the mechanism of their involvement in human brucellosis immunity.Statistical methods: the data of normal use((?)±S)description,skewness data using P50(P25,P75)description.Counting data is described using constituent ratios.One-way analysis of variance or Kruskal-Wallis rank sum test were used to compare groups.The Bonferroni method was further used for pairwise comparison and Spearman correlation analysis was used to clarify the correlation.Results: TLR9 expression level was 1.39(2.12,1.60)in acute stage,1.02(0.82,1.39)in chronic stage and 0.78(0.54,0.96)in healthy control.The expression level of My D88 was0.84(0.64,0.98)in acute stage,0.64(0.38,0.73)in chronic stage and 0.58(0.51,0.63)in healthy control.The expression level of NF-κb was 0.81(0.60,0.94)in acute stage,0.51(0.32,0.62)in chronic stage,and 0.48(0.33,0.65)in healthy control.The expression level of IRF-7was 0.98(0.80,1.10)in acute stage,0.85(0.56,1.00)in chronic stage and 0.74(0.55,0.98)in healthy control.The relative expression levels of TLR9,My D88,NF-κb and IRF-7 in peripheral blood of acute and chronic brucellosis patients were higher than those of healthy control group.The difference between the acute stage and the healthy control group was statistically significant(P<0.05),while the chronic stage and the control group were not statistically significant(P>0.05).Compared with the acute and chronic phases,the expression of TLR9,My D88,NF-κb and IRF-7 in acute stage were higher and the difference was statistically significant(P<0.05).The expression level of IL-4 in acute stage,chronic stage and healthy control was 21.04±4.49,34.58±4.49 and 30.44±4.84 respectively.The expression level of IL-6 was 51.68±4.2 in acute stage,59.94±14.28 in chronic stage and 37.31±4.46 in healthy control.The expression level of IL-12 in acute stage,chronic stage and healthy control was 33.77±5.23,28.38±6.28 and 27.62±4.50 respectively.The expression level of IFN-γ was 525.29±71.94 in acute phase,464.36±64.70 in chronic phase,and 373.17±73.85 in healthy control.Compared with the healthy control group,the levels of IL-6、IL-12 and IFN-γincreased,while IL-4 decreased in the acute stage,all the differences were statistically significant(P<0.05).In chronic stage,the levels of all the above inflammatory factors were higher than those in control group,but only the expression of IL-12 showed no statistical significance(P>0.05).Compared with the acute stage,the levels of IL-4 and IL-6 increased in chronic stage,while the levels of IL-12 and IL-10 decreased,all with statistical significance(P<0.05).There was no correlation between TLR9 expression and My D88 expression in brucellosis patients(r=0.200,P>0.05).My D88 expression was positively correlated with the expressions of NF-κb and IRF-7(r=0.227,0.246,P<0.05).The expression of NF-κb was positively correlated with the expression of IL-12(r=0.385,P<0.01),negatively correlated with the expression of IL-4(r=-3.69,P<0.01),but not significantly correlated with the expression of IL-6 and IFN-γ(r=0.100,0.215,P>0.05).IRF-7 was negatively correlated with the expression of IL-4(r=-3.60,P<0.01),but not significantly correlated with the expression of IL-6,IL-12 and IFN-γ(r=0.209,0.225,0.125,P>0.05).Conclusions:1.In human brucellosis,TLR9 may be activated to participate in the early immune response.2.In the early stage of infection,other receptors may activate My D88 signaling pathway and participate in the immune response.3.With the prolongation of the disease,the expression of TLR9 and the Myd88 signaling pathway may be inhibited,leading to the weakened bactericidal effect,which is not conducive to the elimination of brucella.4.There are dynamic changes of cytokines in human brucellosis,which may be related to the course of brucellosis.