Effect Of Neutrophil Extracellular Traps On Disease Activity And Severity Of Clinical Symptoms In Patients With Primary Sjogren Syndrome

Background:Neutrophil extracellular traps,as a product formed after neutrophil activation,are involved in the pathogenesis of a variety of diseases.With the in-depth exploration of NETs,the relationship between NETs and autoimmune diseases is slowly emerging.Nowadays,a variety of autoimmune diseases,such as SLE and RA,have been confirmed to involve NETs in terms of disease severity and pathogenesis.Various antibodies and cytokines in the disease can activate neutrophils through different pathways,rupture their envelope,and release network structures(NETs)in which DNA binds to some antibacterial proteins and other substances.This network structure can not only aggravate the damage of tissues and organs,but also further promote the production of antibodies and cytokines.However,the relationship between primary Sjogren syndrome(p SS)and NETs is not well understood,and this study aims to further explore the relationship between NETs and primary Sjogren syndrome severity and disease activity.Methods:1.First,this study collected fresh blood samples from p SS patients and gender-agematched healthy controls and clinical data of the results of this visit from p SS patients who visited and hospitalized during the study,and collected the corresponding p SS patients SS disease activity index(ESSDAI)score,the European Union Against Rheumatism SS patient reporting index(ESSPRI)score,Dry Mouth Scale(SXI)score,Anderson Dysphagia Scale(MDADI)score,Hospital Anxiety and Depression Scale(HADS)score,Oral Health Impact Scale(OHIP-14)score,Eye Disease Score Scale(OSDI)score,Fatigue Profile Scale(Pro F)score,Fatigue Severity Scale(FSS)score are related scales used to assess disease activity and severity in patients with p SS,and statistical scores are performed according to the scoring criteria of each scale.2.Secondly,by immunofluorescence colocalization,Myeloperoxidase,histone H3(H3cit)and DNA,three important components of NETs formation,were stained and collected under an upright fluorescence microscope,and the presence of NETs formed by neutrophils in the peripheral blood of p SS patients was determined.Then,through flow cytometry,enzyme-linked immunosorbent assay(ELISA)and other experimental methods,the content of NETs in the peripheral blood of p SS patients and HC was compared,and the relationship between the content of NETs in patients with Primary Sjogren’s syndrome and healthy controls was further explored.3.Finally,the SPSS 25.0 statistical software was used to analyze the correlation between NETs concentration and ESSDAI score,ESSPRI score,SXI score,MDADI score,HADES score,OHIP-14 score,OSDI score,Pro F score and FSS score in p SS patients.Results:1.Experiments showed,immunofluorescence colocalization proves that NETs form in the peripheral blood of p SS patients and can be observed by humans.Flow cytometry results showed that NETs expression in p SS patients was higher than in healthy controls(15.5±7.38 vs 29.4±8.14,t=-2.949,P=0.016).The results of peripheral blood plasma ELISA showed that the concentration of MPO(5263.84±1225.757 vs 4393.58±1073.841,t=-3.52,P=0.001)in peripheral plasma of p SS patients increased significantly compared with the control group.2 the correlation analysis results show that:(1)The ESSDAI score was positively correlated with NETs formation(r2=0.309,P<0.05),and there were no differences between ESSPRI score,ESR,C-reactive protein(CRP),disease course and NETs formation(r2=0.309;r2=0.099;r2=0.174;r2=0.130;r2=0.324,P>0.05).(2)The concentration of MPO in the patient’s peripheral blood was statistically significant with the patient’s SXI score and MDADI score(r2=0.379;r2=0.361,P<0.05).(3)MPO concentration in peripheral blood and HADES score were statistically significant in terms of anxiety and depression(r2=0.324;r2=0.339,P<0.05).(4)There was no significant significance between the MPO concentration in the patient’s peripheral blood and the patient’s OHIP-14 score,OSDI score,Pro F score and FSS score(r2=0.089;r2=-0.16;r2=0.149;r2=0.197,P>0.05).Conclution:This article not only clarifies the difference in NETs concentrations between p SS patients and healthy controls,but also demonstrates a correlation between NETs concentrations in patients with Sjogren’s syndrome and disease activity,dry mouth,swallowing,anxiety and depression.(1)The concentration of NETs in p SS patients was higher than that in HC group.(2)The concentration of NETs in p SS patients was positively correlated with the disease activity of patients.(3)The concentration of NETs in p SS patients was positively correlated with the degree of dry mouth and dysphagia.(4)The concentration of NETs in p SS patients was positively correlated with anxiety and depression.