Relationship Between NLRP3 Inflammasome And Related Cytokines And First-episode Major Depression Disorder

ObjectiveTo detect NLRP3 inflammasome,Caspase-1,IL-18 and IL-1β in peripheral blood of patients with First-episode major depression disorder(FED),and to analyze whether it is involved in the first stage of Major Depression Disorder(MDD),and further clarify the pathogenesis of FED from the perspective of neuroimmunity.Methods1.Using case-control method,64 patients with FED who met DSM-5 diagnostic criteria were selected as case group,and 40 healthy controls matched with their age,gender,education years and marital status were selected as healthy group.2.Hamilton Depression Scale-24(HAMD-24)was selected as the disease severity evaluation tool to evaluate the baseline disease severity of the subjects,and Life Event Scale(LES)was used to evaluate the mental stress factors.3.The levels of IL-1β,IL-18,Tau protein,neurofilament light(NFL)and mannitol binding lectin(MBL)in plasma were detected by Enzyme Linked Immunosorbent Assay(ELISA);the levels of NLRP3 inflammasome and Caspase-1 in Peripheral Blood Mononuclear Cells(PBMC)were detected by Western blot.4.SPSS21.0 was used for data processing and analysis.The quantitative data that conform to normal distribution were described as mean ± standard deviation(x±s),and those that do not conform to non-normal distribution were represented as median(Inter-quartile range)[M(QR)].Two independent samples t-test was used for the two groups of quantitative data which were consistent with normal distribution and homogeneity of variance;two independent samples rank sum test was used for the two groups of quantitative data which were not consistent with normal distribution.x2 test was used for qualitative data.For correlation analysis,Pearson correlation analysis was used for those with normal distribution,and Spearman correlation analysis was used for those without normal distribution.The difference was statistically significant(P<0.05).Results1.Comparison of the levels of inflammatory factors and neuron damage markers between the two groupsCompare the NLRP3 inflammasome,Caspase-1,IL-18,IL-1,NFL,Tau protein,MBL in the peripheral blood of the two groups of patients in the case group and the healthy group.The plasma levels of IL-18 and IL-1β in the case group were higher than those in the healthy group(z=-3.832、-2.362,P<0.05);the levels of NLRP3 inflammasome,Caspase-1,Tau protein,NFL and MBL in the case group were not significantly different from those in the healthy group(z=-0.236,t=-0.500,z=-0.388、-0.735、-1.687,P>0.05).2.ROC curve analysis of inflammatory factors and neuron damage markersThe ROC curve was established with the presence and absence of depression as state variables,NLRP3 inflammasome,Caspase-1,IL-18,IL-1β,NFL,Tau protein and MBL as test variables.The area under the curve(AUC)of IL-18 and IL-1β were 0.724 and 0.638,respectively,with statistical significance(P<0.05).3.The relationship between inflammatory factors,neuron damage markers and the severity of FED in case groupSpearman rank correlation analysis showed that NLRP3 inflammasome was positively correlated with HAMD-24 score(rs=0.3 12,P<0.05),IL-1 8 was positively correlated with IL-1β,NFL,Tau protein and MBL(rs=0.935、0.443、0.661、0.791,P<0.01);IL-1β was positively correlated with NFL,Tau protein and MBL(rs=0.392、0.571、0.732,P<0.01);MBL was positively correlated with NFL and Tau protein(rs=0.459、0.891,P<0.01);NFL was positively correlated with Tau(rs=0.405,P<0.01).4.Comparison of inflammatory factors and neuron damage markers in patients with different clinical subtypes of FED(1)Comparison of FED patients with and without stress events:According to the Life Event Scale(LES),64 patients with FED were divided into two groups according to whether there were negative life events in one month before the onset of FED.There were 34 patients in stress group,30 patients in non stress group.The levels of NLRP3 inflammasome,Caspase-1,IL-18,IL-1β,Tau protein,NFL and MBL in peripheral blood of stress group and non stress group were compared.It was found that the level of NLRP3 inflammasome in stress group was higher than that in non stress group(z=-2.764;P<0.05),and the level of Tau protein in stress group was lower than that in non stress group(z=-2.416,P<0.05),while there were no significant differences in Caspase-1,IL-18,IL-1β,NFL and MBL between stress group and non stress group Statistical significance(t=1.120,z=-0.242、-0.525、-0.444、-1.318,P>0.05).(2)Comparison of FED patients with or without family history:According to whether they have family history or not,64 patients with FED were divided into two groups,including 15 patients with family history and 49 patients without family history.The levels of NLRP3 inflammasome,Caspase-1,IL-18,IL-1β,Tau protein,NFL and MBL in peripheral blood of patients with and without family history were compared.The results showed that there was significant difference in MBL between patients with family history and patients without family history(z=-2.368,P<0.05);there was no significant difference in the levels of NLRP3 inflammasome,Caspase-1,IL-18,IL-1β,Tau protein and NFL(z=-0.463,t-0.659,z=-1.205、-1.355、-6.740、-0.563,P>0.05).(3)Comparison of FED patients with or without psychotic symptoms:According to the presence or absence of psychotic symptoms,64 patients with FED were divided into two groups,including 27 patients with psychotic symptoms(PD group)and 37 patients without psychotic symptoms(NPD group).The levels of NLRP3 inflammasome,Caspase-1,IL-18,IL-1β,Tau protein,NFL and MBL in peripheral blood of PD and NPD groups were compared.It was found that there was no significant difference in NLRP3 inflammasome,Caspase-1,IL-18,IL-1β,Tau protein,NFL and MBL between PD and NPD groups(t=-0.997、0.693,z=-1.339、-0.999、-0.347、-0.707、-0.918,P>0.05).5.Comparison of inflammatory factors and neuron damage markers in male and female FED patientsAmong 64 FED patients,36 were female,28 were male.The results showed that the levels of IL-18,IL-1β and MBL in female were higher than those in male(z=-2.023、-1.678、-2.368,P<0.05),but there was no significant difference in NLRP3 inflammasome,Caspase-1,NFL and Tau protein(t=1.362,z=-0.467、-1.678、-1.848,P>0.05).Conclusions1.There was a tendency of immune activation mediated by IL-18 and IL-1β in FED.No evidence of NLRP3 inflammasome and Caspase-1 involved in the pathogenesis of FED was found.2.FED with family history may have high-risk inflammatory and immune changes.3.Compared with male patients,female FED has more obvious activation of IL-18,IL-1β,and MBL,suggesting that female patients may have more obvious inflammatory and immune damage.Part II The effect of SSRIs on NLRP3 inflammasome and related cytokines in patients with First-episode major depression disorderObjectTo observe the changes of NLRP3 inflammasome and related cytokines after treatment with selective serotonin reuptake inhibitors(SSRIs)in FED patients,and analyze the relationship between them and related markers of neuron damage,so as to further clarify the mechanism of SSRIs.Methods1.Using the method of self-control before and after treatment,34 patients in the case group(hereinafter referred to as the treatment group)were assessed twice for disease severity and peripheral elbow venous blood collection before treatment and after 6-8 weeks of treatment.Fluvoxamine or sertraline is a therapeutic drug(therapeutic dose is 150-300mg).2.The HAMD-24 was used to evaluate the patients in the treatment group twice before treatment(baseline level)and after treatment(6-8 weekends),and the reduction rate was used as the basis for clinical efficacy judgment.3.The levels of plasma IL-1β,IL-18,Tau protein,NFL and MBL were detected by ELISA before treatment and 6-8 weeks after treatment.The levels of NLRP3 inflammasome and Caspase-1 in PBMC before treatment and 6-8 weeks after treatment were detected by Western-blot.The level of serum CRP before treatment was detected by electrochemiluminescence.4.SPSS21.0 was used for data processing and analysis.Quantitative data whose inflammatory factor levels before and after treatment conform to a normal distribution adopts paired t-test;quantitative data whose difference does not conform to a normal distribution adopts the signed rank sum of paired data Test.when performing correlation analysis,Pearson correlation analysis is used for those that meet the normal distribution,and Spearman correlation analysis is used for those that do not meet the normal distribution.When P<0.05,the difference is statistically significant.Results1.Comparison of inflammatory factors and neuron damage markers before and after treatmentThe levels of NLRP3 inflammasome,Caspase-1,IL-18,IL-1β,Tau protein,NFL and MBL in peripheral blood of the treatment group before and after treatment were compared.The results showed that compared with before treatment,the levels of NLRP3 inflammasome、IL-18 and IL-1β increased significantly after treatment,and the difference was statistically significant(t=-2.663、z=-2.350、z=-3.690,P<0.05);Tau protein level and HAMD-24 total score decreased significantly after treatment,and the difference was statistically significant(z=-2.309,t=9.436,P<0.05);while Caspase-1,NFL and MBL had no significant difference before and after treatment(t=-1.423,z=-1.019、-1.706,P>0.05).2.The relationship between the changes of inflammatory factors and the changes of neuron damage markers before and after treatmentSpearman rank correlation analysis showed that the difference of NLRP3 inflammasome before and after treatment was positively correlated with that of NFL and MBL(rs=0.564、0.494,P<0.01);the difference of IL-1β before and after treatment was positively correlated with that of IL-18(rs=0.496,P<0.01);the difference of Tau protein before and after treatment was positively correlated with that of NFL(rs=0.479,P<0.01);the difference of MBL before and after treatment was positively correlated with NFL and Tau protein(rs=0.474、0.619,P<0.01).3.Comparison of the difference between inflammatory factors and neuron damage markers in patients with effective and ineffective FED before and after treatmentThe patients were divided into an effective group(n=21,of which there were 7 cases with a 50%-70%reduction rate,and 14 cases with a 70%-100%)and an ineffective group(n=13,where the reduction rate was 0%-30%had 4 cases,30%-50%had 9 cases).The difference of NLRP3 inflammasome,Caspase-1,IL-18,IL-1β,Tau protein,NFL,MBL before and after treatment was compared between the two groups,the differences of NLRP3 inflammasome,Caspase-1 and MBL between the two groups were statistically significant(t=-2.101、2.270,z=-2.957,P<0.05).Taking Whether treatment is effective as dependent variables,the difference of NLRP3 inflammasome before and after treatment,Caspase-1 before and after treatment,MBL before and after treatment as independent variables,multivariate logistic regression analysis showed that the influencing factors of different effective levels were:NLRP3 inflammasome before and after treatment(OR=4.668,95%CI:1.018-21.399,P=0.047),Caspase-1 difference before and after treatment(OR=0.204,95%CI:0.046-0.895,P=0.035).4.The effects of baseline CRP level on inflammatory factors and markers of neuron damageAccording to the baseline CRP level,the treatment group was divided into high CRP group(CRP>1mg/L,n=13)and low CRP group(CRP<1mg/L,n=21).(1)Comparison of high CRP group before and after treatment:The levels of NLRP3 inflammasome,Caspase-1,IL-18,IL-1β,MBL,NFL,Tau protein and the total score of HAMD-24 in high CRP group were compared before and after treatment.The results showed that after treatment,NLRP3 inflammasome increased significantly(t=-3.376,P<0.01),NFL and Tau protein decreased significantly(z=-2.132,t=2.398,P<0.05),and the total score of HAMD-24 decreased significantly(z=-2.934,P<0.01).There was no significant difference in Caspase-1,IL-18,IL-1βand MBL before and after treatment(P>0.05).(2)Comparison of low CRP group before and after treatment:The levels of NLRP3 inflammasome,Caspase-1,IL-18,IL-1β,MBL,NFL,Tau protein and the total score of HAMD-24 in the low CRP group before and after treatment were compared.The results showed that the total score of HAMD-24 was significantly lower than that before treatment(t=3.985,P<0.01).There was no significant difference in NLRP3 inflammasome,Caspase-1,IL-18,IL-1β,MBL,NFL and Tau protein levels before and after treatment(P>0.05).Conclusions1.SSRIs had a certain tendency to activate NLRP3 inflammasome,and the activation of NLRP3 inflammasome might be more obvious in the effective group.2.SSRIs may have a more significant effect on the markers of neuron damage in patients with baseline CRP>1..