| Background:SLE is a classical autoimmune disease, which is characterized by a myriad of autoantibodies resulted from disturbances of immune response, leading to multi-organ and multi-organs chronic, inflammatory autoimmune disease. The exact pathogenesis of SLE is complex and still unknown. Disturbance of adaptive immunity in SLE has been extensively addressed, however, the role of innate immunity in pathogenesis of SLE remains largely unknown.Innate immunity which is also known as non-specific immune expressed by cells at the front line of defense against pathogenic microorganism, including macrophages, monocytes, dendritic cells, neutrophils and epithelial cells, play a crucial role as a first line of defense against. The innate immune system utilizes pattern recognition receptors (PRRs) to recognize molecular patterns derived from pathogens and damaged cells. Nucleotide binding and oligomerization domain (NOD) receptor (NLR) is a recently identified PRR family member which exists in the cell and sense intracellular danger signals. NOD-like receptor family, pyrin domain containing3and1(NLRP3and NLRP1), two important members of the Nod-like receptor family, can recruit the adaptor ASC, and mediates caspase-1activation and further production of mature IL-1β via assembly of the inflammasome platform in response to various pathogen-derived factors as well as danger-associated molecules. However, despite the potent role of NLRP3and NLRP1inflammasomes in preventing autoimmune disease, their roles in SLE have not been studied extensively. To further study the function of NLRP3and NLRP1inflammasomes in the pathogenesis of SLE and to provide new theoretical basis for immune intervention, we study NLRP3, NLRP1inflammasomes mRNA expression levels in peripheral blood mononuclear cells and granulocytes of SLE patients and healthy controls, explore the association between NLRP3/NLRP1inflammasomes and SLE disease activity, and detect NLRP3/NLRP1mRNA level between pre-treatment SLE patients and post-treatment.Objective:To explore relevant genes expression of NLRP3, NLRP1inflammasome signalling pathway and its association with clinical manifestations in peripheral blood mononuclear cells and granulocytes from SLE patients and healthy controls; to explore the possible role of NLRP3,NLRP1inflammasome signalling pathway in peripheral blood mononuclear cells and granulocyte of SLE pathogenesis.Methods:1.Peripheral blood and was obtained from39SLE patients and31healthy controls.2.Extracted mRNA from the peripheral blood mononuclear cells and granulocyte using Trizol, and then reverse transcription it for cDNA. Real-time PCR technique was used to detect the gene expressions of NLRP3, NLRP1, ASC, Caspase-1, IL-1β mRNA between SLE patients and normal controls. Correlmion analysis was performed between NLRP3, NLRP1, ASC, Caspase-1, IL-1βmRNA expression in peripheral blood mononuclear cells, granulocytes and SLEDAI, ds-DNA, AnuA with Graphpad Prism5.0software.3.The mRNA expression of NLRP3, NLRP1, ASC, Caspase-1, IL-1β was detected before and after glucocorticoid therapy(60mg/d).4.Immunohistochemistry technique was used to detect the gene expressions of NLRP3,ASC and IL-1β between Lupus nephritis (LN) organization and normal kidney tissue.Results:1.Expression of NLRP3, NLRP1inflammasomes from SLE patients and healthy control1.1Expression of NLRP3, NLRP1, ASC, Caspase-1, IL-1βmRNA in PBMC and granulocytes from SLE patients and healthy control. ①Expressions of NLRP3(p=0.0009), NLRP1(p=0.0001), Caspase-1(p=0.03), IL-1β (p=0.0357) mRNA were significantly lower in PBMC from SLE patients than those from healthy controls while no significant difference of ASC expression from SLE patients and healthy controls with RT-qPCR...②Expression of IL-1β mRNA in granulocytes were significantly lower in granulocytes from SLE patients than those from healthy controls (p=0.0408)1.2Expression of NLRP3, NLRP1, ASC, Caspase-1, IL-1βmRNA in LN patients and healthy controls.NLRP3inflammatory were expressed in LN renal tissue and were collected with type of renal pathology type,which suggesting that NLRP3signalling pathway was involved in LN pathogenesis.2. Correlation between the expression of NLRP3, NLRP1, ASC, Caspase-1, IL-1βResult shows that positive correlation between NLRP3and Caspase-1(r=0.7405,P<0.0001), NLRP3and IL-1β (r=0.6085,P=0.0053), NLRP1and Caspase-1(r=0.8010,P<0.0001), NLRP1and IL-1β (r=0.4745,P=0.0053), Caspase-1and IL-1β (r=0.5175, P=0.0010), NLRP3and NLRP1(r=0.7055,P<0.0001) in SLE patient.3. Correlation between expression level of NLRP3, NLRP1inflammasomes and disease activity3.1Correlation between the expression of NLRP3, NLRP1inflammasomes and SLEDAIExpression of NLRP3mRNA are significantly negatively correlated with SLEDAI (r=-0.3959, P=0.0205). Furthermore, other inflammasome components also have a significantly negative correlation between expression level and SLEDAI score, including NLRP1(r=-0.4113, p=0.0174), ASC (r=-0.4860,p=0.0087), and Caspase-1(r=-0.3989, p=0.0215). No correlation were found of IL-1β mRNA expression between SLE patients and healthy controls. No correlation were found of NLRP3, NLRP1inflammasomes and SLEDAI in granulocytes between SLE patients and healthy controls, P>0.05.3.2Correlation between the expression of NLRP3/NLRP1inflammasomes and anti-dsDNAIt showed that there was a significant negative correlations between the anti-dsDNA antibodies and the expression level of NLRP3(r=-0.5493, p=0.0011,), NLRP1(r=-0.4077,p=0.0281) and Caspase-1(r=-0.4002, p=0.0284) in SLE patients. No correlation were found in ASC and IL-1βmRNA expression between SLE patients and healthy controls. No correlation were found of NLRP3, NLRP1inflammasomes mRNA expression and anti-dsDNA in granulocytes between SLE patients and healthy controls, P>0.05.3.3Correlation between the expression of NLRP3, NLRP1inflammasomes and anti-AunAAnalysis of the correlations of inflammasomes expression and anti-AnuA also showed significant negative correlations between anti-AnuA and NLRP1expression (r=-0.3357, p=0.0486, Fig.4D), Caspase-1expression (r=-0.3699, p=0.0372, Fig.4E), IL-1β expression (r=-0.3544, p=0.0465, Fig.4F). No correlation were found of NLRP3, ASC inflammasomes mRNA expression and anti-AunA in granulocytes between SLE patients and healthy controls, P>0.05.4. NLRP3, NLRP1inflammasomes mRNA level between pre-treatment SLE patients and post-treatment.Expression of NLRP3, NLRP1inflammasomes were increased after glucocorticoid therapy.There was a significant differences in expression levels of NLRP3(p=0.0237), NLRP1(p=0.0436), Caspase-1(p=0.0252) and IL-1β(p=0.0148) between pre-treatment SLE patients and post-treatment ones. No significant difference of ASC mRNA expression between pre-treatment SLE patients and post-treatment ones were found.Conclusion:1.The expression of NLRP3, NLRP1inflammasomes were significantly downregulated in PBMCs and NLRP3expression were significantly upregulated in granulocytes in SLE patients, and regular glucocorticoids treatment significantly corrected these inflammasomes deregulation.which prompt that dysregulated expression of NLRP3, NLRP1inflammasome in PBMC and granulocyte of SLEpatients was involved in pathogenesis of SLE.2.Expressions of NLRP3, NLRP1inflammasomes were negatively correlated with the SLE disease activity index (SLEDAI, anti-dsDNA, anti-AnuA), which suggested an important role for inflammasomes in SLE pathogenesis.3. NLRP3, ASC, IL-1β inflammatory were expressed in LN renal tissue and were collected with type of renal pathology type,which suggesting that NLRP3-ASC signalling pathway was involved in LN pathogenesis. Background:Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease, which is characteristic of limb joint involvement.The cause of RA is unknown and his course and outcome were differences.It has confirmed that RA is the result of the role of genes, environment.Innate and adaptive immunity are both involved in the pathogenesis of RA.NOD-like receptor family, pyrin domain containing3and1(NLRP3and NLRP1), two important members of the Nod-like receptor family, can recruit the adaptor ASC, and mediates caspase-1activation and further production of mature IL-1β via assembly of the inflammasome platform in inflammatory response and innate immune response. The activation mechanism of NLRP3, NLRP1is mainly concentrated in the innate immune cells which is still rarely studied in RA.Objective:To explore relevant expression of NLRP3, NLRP1inflammasome constituent in peripheral blood mononuclear cells and granulocytes from RA patients and healthy controls; To analysis the relation between expression of NLRP3, NLRP1inflammasome constituent and clinical manifestations to explore the possible role of NLRP3, NLRP1inflammasome in RA signalling pathway.Methods:1.Peripheral blood and granulocyte were obtained from36SLE patients and30healthy controls.2. Real-time PCR technique was used to detect the gene expressions of NLRP3、 NLRP1, ASC, Caspase-1, IL-1β mRN A between RA patients and normal controls. Correlation analysis was performed between NLRP3. NLRP1, ASC, Caspase-1, IL-1β mRNA expression and RF,CCP,ESR with Graphpad Prism5.0software.Results:1.Expression of NLRP3, NLRP1inflammasomes1.1Expression of NLRP3, NLRP1inflammasomes comment in PBMC and granulocytes from RA patients and healthy control.Expressions of NLRP1(p=0.0457),Caspase-1(p=0.0273) were significantly lower in PBMC from RA patients than those from healthy controls while no significant difference of NLRP3, ASC,IL-1β expression from RA patients and healthy controls.1.2Expression of NLRP3(p=0.0489), ASC (p=0.0064) and Caspase-1(p=0.0249) in granulocytes were significantly lower in granulocytes from RA patients than those from healthy controls.3. Correlation between expression level of NLRP3, NLRP1inflammasomes and CCP, RF, ESRExpression of NLRP1was significantly negatively correlated with CCP (r=-0.3674, P=0.0354) and RF (r=-0.4189,P=0.0332). Expression of IL-1β was significantly negatively correlated with RF (r=-0.3948, P=0.0340).Conclusion:The expression of NLRP3, NLRP1and Caspase-1inflammasomes were significantly downregulated in PBMCs and granulocytes of RA patients, which prompt that NLRP3, NLRP1inflammasome was involved in pathogenesis of RA. Expressions of NLRP1inflammasomes were negatively correlated with CCP and RF, which suggested that NLRP1inflammasomes may be a new therapeutic target in RA. |
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