| Background:Systemic lupus erythematosus(SLE)is a type of systemic autoimmune disease,commonly found in young women of childbearing age,which mainly characterized by the activation of T and B cell,formation of numerous different autoantibodies,immune complex(IC),and damage to multiple tissues and organs.Its etiology and pathogenesis are not yet clear.The mechanisms involved in the pathogenesis of SLE include genetic factors,environmental factors,hormone levels,and immune regulation.The immunological mechanism of SLE onset almost covers the entire immune system network,in which the innate and adaptive immune systems play very important roles.As the human body's first immune barrier system,the innate immune system recognizes pathogen-associated molecular patterns(PAMP)through pattern recognition receptors(PRRs)to play a role in eliminating foreign pathogens and activating adaptive immune responses.In the currently discovered PRRs,some NOD like receptors(NLRs)that can be activated to form intracellular high-molecular-weight protein complexes referred to as inflammasome.The inflammasome activates cysteinyl aspartate-specific proteinase(Caspase-1),thereby prompting the production and activation of inflammatory cytokines proinflammatory interleukin-18(IL-18)and interleukin-1 beta(IL-1 ?),which take participate in the process of inflammation and immune response.The reported inflammasomes mainly include:ice-protease-activating factor(IPAF),absent in melanoma 2(AIM2),Nod-like receptor protein 1(NLRP1)and Nod-like receptor protein 3(NLRP3),of which the NLRP3 is the most widely studied inflammasome.In our previous studies,we found that mRNA and protein expression of NLRP3 inflammasome components(ASC,NLRP3,Caspase-1)and IL-1? were lower expressed in peripheral blood mononuclear cells(PBMC)of SLE patients compared with healthy controls,and its expression level was negatively correlated with disease activity related indicators(SLEDAI,anti-ds-DNA antibody,anti-AnuA antibody).It indicates that the inactivation of NLRP3 inflammasome is involved in the pathogenesis of SLE.It is speculated that NLRP3 inflammasome may be a protective factor of SLE,but how it plays a role has not been fully elucidated.Interferon(IFN),known as an cytokine whose chemical essence is protein,which has antiviral infection,anti-proliferation,effective regulation of cell division and immune activation.It binds to the corresponding receptor IFNAR on the surface of the cell membrane,which in turn activates a non-receptor tyrosine protein kinase JAK(Janus Kinase)and causes the dimerization of receptor molecules.The kinase JAK catalyzes the phosphorylation modification of STAT(Signal transducers and activators of transcription)proteins bound to the receptor.The activated STAT protein enters the nucleus as a dimer and binds to target genes to regulate gene transcription.The JAK-STAT signaling pathway has a wide range of functions and plays an important role in immune response and regulation.In recent years,several studies have shown that there is high expression of type I and III interferon in the serum of SLE patients,and it is closely related to SLE disease activity and severity.Therefore,we speculate that there is a mechanism by which interferon negatively regulates the NLRP3 inflammasome in SLE patients,but the specific mechanism has not been elucidated.Objective:To investigate the expression levels of IFN in SLE and the role and clinical significance of interferon negatively regulating the NLRP3 inflammasoma signaling pathway in the pathogenesis of SLE.Methods:The study recruited 42 SLE patients(SLE group)treated from the Department of Rheumatology and Immunology in Provincial Hospital Affiliated to Shandong University from December 2018 to december 2019 and 30 healthy people who were admitted to the hospital's physical examination center(Healthy control group)during the same period.Real-time Polymerase Chain Reaction(RT-PCR)was used to detect the mRNA expression of IFN-?,IFN-?,IFN-?1,NLRP3 inflammasome components(ASC,NLRP3,Caspase-1)and its downstream cytokines(IL-18,IL-1 p)in PBMCs of two groups of subjects.The serum levels of IFN-a,IFN-?,IL-18,IL-1? were measured by Human Magnetic Luminex Assay,and IFN-?1 were measured by enzyme-linked immunosorbent assay(ELISA).The expression levels of the above indicators and their correlation with clinical manifest,laboratory indicators,disease activity evalution and other data were analyzed.Results:1.The results of RT-PCR experiments showed that compared with the healthy control group,the IFN-?,IFN-?,IFN-?1,NLRP3,and IL-1 ? mRNA expression levels were lower in the PBMCs of the SLE patients(P<0.0001;P<0.0001;P<0.0001;P=0.006 and P=0.0007,respectively);The mRNA expression levels of ASC and IL-18 were higher in SLE patients,but the difference was not statistically significant(P=0.2728 and P=0.8078;respectively).There was no statistically significant of the mRNA expression level of Caspase-1 between the SLE group and the healthy control group(P=0.1741)2.ELISA and Luminex test results showed that compared with healthy controls,serum levels of IFN-?,IFN-?,IFN-?1,IL-18 and IL-1? were significantly higher in SLE patients(P=0.0003;P=0.0371;=0.0126;P<0.0001;P=0.0089;respectively).3.The mRNA expression levels of IFN-?,IFN-?,IFN-?1,ASC,Caspase-1,NLRP3,IL-18,and IL-1? in PBMCs between the newly diagnosed SLE group and the treated SLE patients group were not significantly different(P=0.077,0.276,0.59,0.98,0.159,0.179,0.348,0.307,respectively).4.Compared with the newly diagnosed SLE group,the serum level of IFN-? in the treated SLE group was lower(P=0.018),and the serum levels of IFN-?,IFN-?1,IL-18 and IL-1? was not significantly different between the two groups(P=0.125;P=0.276;P=0.06 and P=0.291;respectively)5.There was no significant difference in the mRNA expression levels of IFN-?,IFN-?,IFN-?1,NLRP3 inflammasome components and downstream factor between the SLE patients without renal damage and patients with lupus nephritis(0.4869±0.6729 vs 0.6555±0.8109(P=0.22)for IFN-?,0.7963±0.6131 vs 0.8543±0.3290(P=0.04)for IFN-?,0.6673±0.7690 vs 0.4704± 0.3450(P=0.953)for IFN-?1,0.3914 ± 0.1553 vs 0.4619±0.2427(P = 0.555)for ASC,0.8063± 0.3089 v 0.4344(P?0.734)for Caspase-1,0.4679±0.6313 vs 0.2952±0.1729(P=0.768)for NLRP3,0.7942 ±0.7023 vs 0.6867±0.3172(P=0.913)for IL-18,0.5349± 1.1838 vs 0.2251 ± 0.3171(P=0.639)for IL-1?.6.Compared with the SLE patients without renal damage,patients with lupus nephritis showed higher serum levels of IFN-? and IFN-?3:2.3692± 1.8534 vs 8.5890±6.1574(P=0.006)for IFN-?,7.4972±15.6107 vs 88.6778±157.9863(P=0.006)for IFN-?;The expression levels of IL-18 and IL-1? were higher,but the difference was not statistically significant(557.6416±234.5273 vs 659.7910±499.0469(P=0.745)for IL-18,3.6400± 0.6842 vs 44.5550± 97.8658(P=0.147)for IL-1?;and serum levels of IFN-?1 was lower in patients with lupus nephritis,but the difference was not statistically significant:145.9535± 46.6974 vs 125.2203± 28.5743(P=0.14).7.The mRNA expression levels of IFN-?,IFN-?,IFN-?1,NLRP3 inflammasome components and their downstream cytokines in SLE patients with different disease activity showed that only the mRNA expression level of Caspase-1 was higher in the mildly active patients than the severely active group(0.9359±0.3899 vs 0.6196±0.1350,P=0.027),while there were no significant differences of the mRNA expression levels of IFN-?,IFN-?,IFN-?1,ASC,NLRP3,IL-18,and IL-1? in SLE patients with different diseases activity levels.8.In the serum of SLE patients with different disease activity groups,IFN-? and IFN-?were highly expressed in the severely active group(1.6775± 0.4757 vs 9.0738± 6.1409(P=0.05)for IFN-?,2.0365± 0.6734 vs 9.0738±6.1409(P=0.009)for IFN-?,3.6708±3.8431 vs 9.0738±6.1409(P=0.044)for IFN-?;2.4875 ± 0.5950 vs 100.8529 ±178.6054(P=0.01)for IFN-?,10.5641 ± 21.1690 vs 100.8529 ± 178.6054(P=0.014)for IFN-?,10.9615 ± 23.3644 vs 100.8529± 178.6054(P=0.024)for IFN-?,and there was no significant difference of IFN-?1,IL-18 and IL-1? in the serum of SLE patients with different disease activity groups.9.The results of correlation analysis showed that there was a positive correlation of mRNA levels between the IFN-? and IFN-? in PBMCs in the SLE patients(r=0.4371,P=0.0016);and there was no correlation between the mRNA expression levels of IFN-?and IFN-?1,ASC,Caspase-1,NLRP3,IL-18,IL-1?(r=0.2280,0.0065,0.0751,0.0647,0.0580,-0.2138,P=0.1464,0.9675,0.6366,0.6841,0.7223,0.1853;respectively).There was no correlation between the mRNA expression levels of IFN-?and IFN-?1,ASC,Caspase-1,NLRP3,IL-18,IL-?(r= 0.2288,-0.1390,0.0620,0.2813,-0.0175,-0.0854,P=0.1450,0.3801,0.6967,0.071 1,0.9144,0.6004;respectively).There was no correlation between the mRNA expression levels of IFN-?1 and ASC,Caspase-1,NLRP3,IL-18,IL-1?(r=0.1739,0.2123,0.0006,-0.2066,-0.1423,P=0.2707,0.1770,0.9972,0.2010,0.3810;respectively).The mRNA expression level of NLRP3 was positively correlated with IL-1?(r=0.4749,P=0.002),but has no correlation with the mRNA expression levels of ASC,Caspase-1,and IL-18(r=-0.0295,0.1734,-0.1087,P=0.8529,0.2720,0.5042;respectively).There was a positive correlation between ASC and Caspase-1 mRNA expression level(r=0.6904,P<0.0001),but not correlated with IL-18,IL-1? mRNA levels(r=0.1854,0.0273,P=0.2520,0.8672;respectively).There was no significant correlation between Caspase-1 mRNA expression levels and IL-1 8,IL-1?(r=-0.0028,0.1977,P=0.9865,0.2214;respectively);so as IL-18 and IL-1? mRNA expression levels(r=0.1177,P=0.4696).10.Correlation analysises of the mRNA expression levels of IFN-?,IFN-?,IFN-?1,NLRP3 inflammasome components and their downstream cytokine with clinical datas(SLEDAI,anti-dsDNA,anti-AnuA,ESR and CRP):10.1 The mRNA expression levels of IFN-? were negatively correlated with ESR and CRP(r=-0.3256,-0.3675,P=0.0354,0.0167;respectively),and had no correlation with SLEDAI,anti-dsDNA,and anti-AnuA antibody(r=0.0238,-0.1778,0.1052,P=0.8812,0.2600,0.5075;respectively).10.2 The mRNA expression levels of IFN-? were negatively correlated with ESR and CRP(r=-0.4128,-0.3332,P=0.0066,0.0311,respectively),and had no correlation with SLEDAI,anti-dsDNA,and anti-AnuA antibody(r = 0.0545,-0.0010,0.1144,P=0.73 16,0.9930,0.4706;respectively).10.3 The mRNA expression levels of IFN-?1 were negatively correlated with CRP(r=-0.3486,P=0.0237),and had no correlation with SLEDAI,anti-dsDNA,anti-AnuA and ESR(r=-0.0952,-0.0869,-0.0404,-0.1653,P=0.5489,0.5842,0.7997,0.2955;respectively).10.4 There was no correlation between the mRNA expression levels of NLRP3 with SLEDAI,anti-ds-DNA,anti-AnuA,ESR and CRP(r=-0.0228,0.0034,0.0803,-0.2083,-0.1408,P=0.8859,0.9829,0.6131,0.1857,0.3740;respectively).10.5 There was no correlation between the mRNA expression levels of ASC with SLEDAI,anti-ds-DNA,anti-AnuA,ESR and CRP(r=-0.1331,-0.1377,-0.2149,-0.0587,-0.2194,P=0.4008,0.3905,0.1717,0.7120,0.1626;respectively).10.6 The mRNA expression levels of Caspase-1 were negatively correlated with SLEDAI(r=-0.3566,P=0.0205),and had no correlation with anti-dsDNA,anti-AnuA,ESR and CRP(r=-0.2707,-0.2687,-0.0701,-0.1494,P=0.0830,0.0853,0.6594,0.3451;respectively)10.7 There was no correlation between the mRNA expression levels of IL-18 with SLEDAI,anti-ds-DNA,anti-AnuA,ESR and CRP(r = 0.1039,-0.1843,-0.1082,-0.0235,-0.0121,P=0.5234,0.2550,0.5062,0.8857,0.9409;respectively).10.8 There was no correlation between the mRNA expression level of IL-1? with SLEDAI,anti-ds-DNA,anti-AnuA,ESR and CRP(r=-0.0613,-0.1093,-0.0330,-0.0931,-0.1371,P=0.7069,0.5020,0.8398,0.5677,0.3987;respectively).11.The correlation analysis of ELISA test results showed that the serum levels of IFN-?were positively correlated with IFN-?,IL-18,and IL-1?(r=0.5820,0.3346,0.751 1,P<0.0001,P=0.0303,0.0197;respectively),and had no correlation with IFN-?1(r=-0.0261,P=0.8694).Serum levels of IFN-? were positive correlated with IL-1?(r=0.9489,P<0.0001),and had no correlation with IFN-X1,IL-18(r=-0.0762,0.2887,P=0.6360,0.0672;respectively).There was no correlation between the serum levels of IFN-?1 and IL-18,IL-1?(r = 0.1539,-0.2979,P=0.3306,0.4362;respectively),so as IL-18 and IL-1? serum levels(r=-0.3660,P=0.3326).12.ELISA test results correlation analysis showed that the serum levels of IFN-? were positively correlated with anti-AnuA,SLEDAI,anti-SM,and anti-rRNP(r=0.3160,0.5303,0.4850,0.4033;P=0.0415,0.0003,0.0011,0.0081;respectively),and were inversely associated with C3 and C4(r=-0.5045,-0.5491,P=0.0007,0.0002;respectively).Serum levels of IFN-? was positively correlated with anti-AnuA,SLEDAI,24-hour urine protein quantification,and anti-rRNP(r=0.3297,0.5589,0.3887,0.3620;P=0.0353,0.0001,0.012,0.02;respectively),and had negative correlation with C3,C4(r=-0.4526,-0.4724,P=0.003,0.0018;respectively).Serum levels of IFN-?1 was positively correlated with C3 and CRP(r=0.3237,0.3391;P=0.0365,0.028;respectively).Serum levels of IL-18 was positively correlated with IgE(r=0.3067,P=0.0482);serum levels of IL-1 ? was positively correlated with SLEDAI and IgM(r=0.7095,0.8463,P=0.0323,0.004;respectively)?and negatively correlated with C4(r=-0.8652,P=0.0026).Conclusions:1.There are differences in the expression of type ? and type ? IFN in the transcription and translation levels of patients with SLE.It is closely related to the immune response of SLE and related to the disease activity of SLE.Type ? IFN is involved in the pathogenesis of LN.2.In PBMC of SLE patients,IFN and NLRP3 inflammatory bodies are both lowly expressed.IFN may have a negative activation effect on NLRP3 inflammatory bodies,but IFN does not rely solely on NLRP3 inflammatory body signals during the pathogenesis of SLE.Regulation of pathways.3.The expression of Caspase-1 in PBMC of SLE patients is positively correlated with ASC,and there is no obvious correlation with IFN-?,IFN-?,IFN-?1,NLRP3.In addition to the regulation of body signaling pathways,there may be other signaling pathways that regulate the expression of Caspase-1.4.The expression of NLRP3 in PBMC of SLE patients is positively correlated with IL-1 ?,the expression of serum IFN-? is positively correlated with the expression of IL-18 and IL-1 ?,and the expression of IFN-? is positively correlated with IL-1 ?;IL-18 The mediation of IL-1? maturation and activation may be related not only to the regulation of the NLRP3 inflammatory corpuscle signaling pathway,but also to the type? interferon. |
PDF Full Text Request