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Background: Interleukin-34 (IL-34) is an important immunomodulatory factor that plays a crucial role in a variety of autoimmune diseases. Aims: To investigate the expression of IL-34 in primary biliary cholangitis (PBC) and its influence on intrahepatic inflammation and bile duct damage. Methods: Liver tissues were obtained from 26 PBC patients and 10 hepatic hemangioma patients without PBC. Expression and localization of IL-34 were detected by immunohistochemistry. In animal experiment, Poly I:C intraperitoneal injection was used to construct PBC model in wild-type and IL-34-knockout C57BL/6 mice (WT-PBC group and IL-34KO-PBC group). Subsequently, the intrahepatic inflammation and bile duct damage were evaluated pathologically, and the expressions of IL-34 and associated cytokines in liver tissues were detected by real-time PCR and Western blotting. Results: Expression of IL-34 in liver tissues of PBC patients and PBC model mice was significantly higher as compared with those of the controls (all P<0.05). No morphological changes in hepatic pathological evaluation were observed in IL-34KO mice receiving intraperitoneal saline injection. In IL-34KO-PBC mice, the portal area inflammation and biliary epithelial cell damage were more severe than those in WT-PBC mice (all P<0.05). Expressions of proinflammatory cytokine interleukin-1β (IL-1β) and monocyte chemotactic protein-1 (MCP-1) in liver tissues of IL-34KO-PBC mice were significantly increased than those of WT-PBC mice, whereas expressions of antiinflammatory cytokine IL-10 and CD163, the surface marker of M2 macrophages, were significantly reduced (all P<0.05). Conclusions: IL-34 expression is increased in liver tissues of PBC patients and animals. It might reduce the portal area inflammation and bile duct damage via modulating cytokines expression and driving macrophages polarization to the M2 phenotype. IL-34 might act as a self-rescue factor which negatively regulates hepatic immune microenvironment and prevents disease progression.
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SUMMARY OBJECTIVE: The role of interleukins, such as IL-17 and IL-34, in the pathogenesis of autoimmune diseases has been established in the literature. In the current study, we aimed to identify the concentrations of IL-17 (IL-17A, IL-17F) and IL-34 in the cerebrospinal fluid (CSF) of patients with chronic inflammatory demyelinating polyneuropathy (CIDP) and acute inflammatory demyelinating neuropathy (AIDN). METHODS: We included in this study 8 patients with CIDP (none of them receiving immunomodulatory or immunosuppressant therapy), 7 patients with Guillain-Barre syndrome (GBS, AIDN), and 7 control subjects. The CIDP and AIDN diagnoses were made by clinical evaluation and electrophysiological investigations according to international criteria. CSF samples were obtained appropriately, and the levels of IL-17A, IL-17F, and IL-34 were measured by ELISA kits. RESULTS: The concentrations of IL-17A, IL-17F, and IL-34 were higher in those with CIDP and AIDN compared to the controls (p=0.005, p=0.01, and p=0.001, respectively). While IL-34 levels were significantly higher in AIDN patients than in CIDP patients (p=0.04), there were no significant differences between the AIDN and CIDP groups with regard to the levels of IL-17A and IL-17F (p=0.4 and p=0.2, respectively) CONCLUSION: Our results indicate that IL-17A, IL-17F, and IL-34 levels may have a role in CIDP and AIDN. Furthermore, the difference in the IL-34 levels of patients with AIDN and CIDP may indicate an important difference between the pathogenesis of these two sets of the disease.
RESUMO OBJETIVO: O papel das interleucinas, como IL-17 e IL-34, na patogênese da doença auto-imune foi estabelecido na literatura. No presente estudo, objetivamos identificar as concentrações de IL-17 (IL-17A, IL-17F) e IL-34 no líquido cefalorraquidiano (LCR) de pacientes com polineuropatia desmielinizante inflamatória crônica (CIDP) e neuropatia desmielinizante inflamatória aguda (AIDN). MÉTODOS: incluímos neste estudo 8 pacientes com CIDP (nenhum deles recebendo terapia imunomoduladora ou imunossupressora), 7 pacientes com síndrome de Guillain-Barre (GBS, AIDN) e 7 indivíduos controle. Os diagnósticos CIDP e AIDN foram feitos por avaliação clínica e investigações eletrofisiológicas de acordo com critérios internacionais. As amostras de LCR foram obtidas adequadamente e os níveis de IL-17A, IL-17F e IL-34 foram medidos através de kits ELISA. RESULTADOS: As concentrações de IL-17A, IL-17F e IL-34 foram maiores naqueles com CIDP e AIDN em comparação aos controles (p = 0,005, p = 0,01 ep = 0,001, respectivamente). Enquanto os níveis de IL-34 foram significativamente mais altos nos pacientes com AIDN do que nos pacientes com CIDP (p = 0,04), não houve diferenças significativas entre os grupos com AIDN e CIDP em relação aos níveis de IL-17A e IL-17F (p = 0,4 ep = 0,2, respectivamente) CONCLUSÃO: Nossos resultados indicam que os níveis de IL-17A, IL-17F e IL-34 podem ter um papel no CIDP e no AIDN. Além disso, a diferença nos níveis de IL-34 de pacientes com AIDN e CIDP pode indicar uma diferença importante entre a patogênese desses dois conjuntos de doenças.
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Humans , Polyradiculoneuropathy, Chronic Inflammatory Demyelinating , Enzyme-Linked Immunosorbent Assay , Interleukins , Interleukin-17 , Guillain-Barre SyndromeABSTRACT
Objective:To investigate the effects of interleukin-34(IL-34)on the expression of CC chemokine ligand 28 (CCL28) by fibroblast-like synoviocytes(FLS) in rheumatoid arthritis(RA) patients.Methods:FLS were isolated from 6 RA patients and stimulated with IL-34,IL-34 receptor antagonist /IL-34,inhibitors of signaling pathway/IL-34 in vitro respectively.CCL28 mRNA expression was measured by reverse transcription polymerase chain reaction(RT-PCR).The level of CCL28 in the supernatant of RA FLS culture was detected by enzyme linked immunosorbent assay(ELISA).Statistical analysis between groups was performed by t test.Results:Compared with unstimulated FLS,CCL28 expression was increased obviously in IL-34-stimulated group(P<0.05).The level of CCL28 was significantly decreased when anti-IL-34 antibody was added into IL-34-administrated RA FLS(P<0.05).While after adding of nuclear factor κB(NF-κB) and mitogen-activated protein kinase38(p38 MAPK) signaling pathway inhibitors into the cell culture system,CCL28 expression was remarkably reduced (P<0.05).Conclusion: The secretion of CCL28 by RA FLS can be promoted by the binding of IL-34 with its specific receptor via the activation of NF-κB and p38 MAPK signaling pathways,which suggests that CCL28 might be involved in the pathogenesis of RA.
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Objective:To investigate the serum level of IL-34 in patients with acute phase Henoch-Schoenlein purpura ( HSP) and its potential significance .Methods:38 HSP patients were recruited ,together with 21 healthy volunteers as control .The serum level of IL-34,IL-6 and TNF-αwere determined using enzyme-linked immunosorbent assay (ELISA) or chemiluminescence.The correlation analysis was performed to explore the correlation of IL-34 with clinical parameters and the serum level of IL-6,TNF-α.Results:The se-rum levels of IL-34,IL-6 and TNF-αin HSP patients were significantly higher as compared to the control group (P<0.001).The serum level of IL-34 was positively correlated with that of high sensitive C-reactive protein ( hs-CRP) and IL-6 ( r=0.453 ,P=0.004 and r=0.469,P=0.003,respectively).Conclusion:IL-34 might be involved in the pathogenesis of HSP ,and the potential mechanisms of ac-tion is associated with the aberrant expression of pro-inflammatory factors in HSP ,which promote the development of inflammation in the vascular system.
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Objective To investigate and analyze the changes of serum interleukin-34(IL-34),rheumatoid factor(RF) and nuclear factor κB receptor activator ligand(RANKL) in patients with rheumatoid arthritis Its relationship with osteoporosis.Methods A total of 40 patients suffering from rheumatoid arthritis in the hospital between January 2016 and January 2017 were included in the study group,and 40 healthy subjects who underwent the same period of physical examination at the same time were enrolled as controls.By enzyme-linked immunosorbent assay(ELISA) were detected in 2 groups of serum IL-34 and RANKL levels were detected by latex agglutination assay of serum RF levels in 2 groups,and routine testing in patients with bone mineral density,erythrocyte sedimentation rate(ESR),C reactive protein(CRP) and anti CCP antibody.And the modified DAS28 score of disease activity of rheumatoid arthritis evaluation.Results The levels of serum IL-34,RF and RANKL in the study group were significantly higher than those in the control group(P＜0.05).The incidence of osteoporosis in the study group was significantly higher than that in the control group(P＜0.05).The levels of IL-34,RF and RANKL in the osteoporosis patients were significantly higher than those in the normal group(P＜0.05).The Spearman correlation analysis showed that the serum IL-34 levels were positively correlated with ESR,CRP,anti-CCP and DAS28 (P＜ 0.05).There was a negative correlation between serum IL-34,RF and RANKL levels and bone mineral density(P＜0.05).Conclusion The levels of serum IL-34,RF and RANKL in patients with rheumatoid arthritis were significantly increased,and the above indexes were negatively correlated with bone mineral density.It suggested that IL-34 may be involved in osteoporosis.
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Objective To investigate and analyze the changes of serum interleukin-34(IL-34),rheumatoid factor(RF) and nuclear factor κB receptor activator ligand(RANKL) in patients with rheumatoid arthritis Its relationship with osteoporosis.Methods A total of 40 patients suffering from rheumatoid arthritis in the hospital between January 2016 and January 2017 were included in the study group,and 40 healthy subjects who underwent the same period of physical examination at the same time were enrolled as controls.By enzyme-linked immunosorbent assay(ELISA) were detected in 2 groups of serum IL-34 and RANKL levels were detected by latex agglutination assay of serum RF levels in 2 groups,and routine testing in patients with bone mineral density,erythrocyte sedimentation rate(ESR),C reactive protein(CRP) and anti CCP antibody.And the modified DAS28 score of disease activity of rheumatoid arthritis evaluation.Results The levels of serum IL-34,RF and RANKL in the study group were significantly higher than those in the control group(P＜0.05).The incidence of osteoporosis in the study group was significantly higher than that in the control group(P＜0.05).The levels of IL-34,RF and RANKL in the osteoporosis patients were significantly higher than those in the normal group(P＜0.05).The Spearman correlation analysis showed that the serum IL-34 levels were positively correlated with ESR,CRP,anti-CCP and DAS28 (P＜ 0.05).There was a negative correlation between serum IL-34,RF and RANKL levels and bone mineral density(P＜0.05).Conclusion The levels of serum IL-34,RF and RANKL in patients with rheumatoid arthritis were significantly increased,and the above indexes were negatively correlated with bone mineral density.It suggested that IL-34 may be involved in osteoporosis.
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Objective To investigate the effects of interleukin (IL)-34 on the proliferation and chemokines expression of fibroblast-like synoviocytes (FLS) in patients with rheumatoid arthritis (RA).Methods RA FLS were isolated and cultured in the presence or absence of IL-34.FLS proliferation was determined by methyl thiazlyl tetrazoliu method (MTT) method.The levels of chemokines from RA FLS supernatant were detected by protein chip AAH-CYT-G1000 assay.The IL-34 receptor (IL-34R) expression on RA FLS was analyzed by flow cytometry (FCM).Statistical analysis between groups was performed by t test.Results Compared to the control group, the proliferation of IL-34-stimulated RA FLS was obviously increased, and up to the maximum at 72 hours.In addition, the levels of chemokines [epithelial neutrohil-activating peptide 78 (ENA-78), IL-8, growth-related oncogene (GRO), macrophage chemoattractant protein-1 (MCP-1)] on RA FLS supernatant in IL-34 stimulated group were significantly elevated than those in the control group [ENA-78: (397.1±8.2) pg/ml, (54.0±2.9) pg/ml (t=127.61, P＜0.01);IL-8:(2 017±36) pg/ml, (778±102) pg/ml (t=36.67, P＜0.01);GRO: (4 935±160) pg/ml, (2 746±188) pg/ml (t=43.60, P＜0.01);MCP-1: (46 798±1 293) pg/ml, (27 813±2 329) pg/ml (t=22.43, P＜0.01)].IL-34R was highly expressed on the RA FLS.Conclusion IL-34 promotes RA FLS proliferation and chemokines expression, which may be one of the important mechanisms involved in the pathogenesis of RA.
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Objective To preliminarily investigate the levels of interleukin (IL)-1 family and IL-34 in serum of patients with ankylosing spondylitis (AS) and their roles.Methods Serum IL-1 family levels were detected from 6 AS patients and 4 healthy controls by using protein-chip technique.Enzyme-linked immunosorbent assay (ELISA) method was used to detect the levels of serum IL-34 from 65 AS patients and 85 healthy controls and the relationships of serum IL-34 levels and clinical or laboratory features were analyzed.T test and Spearman correlation were used for statistical analysis.Results IL-1Ra [(3302±1352) pg/ml vs (10778±2764) pg/ml]and IL-36Ra [(1363±194) pg/ml vs (3875±996) pg/ml] levels were significantly down-regulated in AS patients compared with that of healthy controls (t=5.363 and 4.289 respectively,both P＜0.05).The levels of IL-1α,IL-18,IL-36α and IL-37 were increased more remarkable in AS patients than in healthy controls (t=-2.532,-5.400,-5.023 and-5.783 respectively,both P＜0.05).Moreover,serum IL-34 levels were elevated more significantly in AS patients than in healthy controls [(169±153) pg/ml vs (54±31) pg/ml,t=6.722,P＜0.01] and were positively correlated with the levels of CRP and ESR.Serum IL-34 levels were markedly up-regulated in human leukocyte antigen (HLA)-B27 positive patients than in HLA-B27 negative patients(P＜0.05).Conclusion Part of IL-1 family and IL-34 may be involved in inflammatory or immunological process of AS.
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Objective To investigate the expression of interleukin (IL)-34/colony stimulating factor(CSF)-1R in the process of transforming growth factor ( TGF)-β1 inducing epithelial-mesenchymal transition ( EMT) of human alveolar epithelial cells A549 cells.Methods A549 cells were cultured in vitro.CCK 8 was used to test the influence of the proliferative rate of A549 cells which were stimulated by TGF-β1 at different concentrations and time points .A549 cells were stimulated by 5μg/L TGF-β1 at 0 hour, the 12th hour, the 24th hour, and the 48th hour.Western blotting was adopted to detect changes of the following proteins: α-smooth muscle actin (α-SMA ) , E-cadherin ( E-Cad ) , matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1(TIMP-1). Real-time PCR was adopted to detect changes of the following genes: IL-34 mRNA, CSF-1R mRNA, MMP-2 mRNA and MMP-9 mRNA.Results TGF-β1 had no significant influence in the proliferation of A 549 cells compared with the control group(P>0.05).TGF-β1(5μg/L)stimulated A549 cells at different time point (0 hour, 12, 24, 48 hours), compared with the control group .The epithelial phenotype E-Cad protein was gradually down-regulated ( P <0.01 ) , while the mesenchymal phenotype α-SMA protein was gradually up-regulated ( P <0.01 ) and the protein of MMP-2 increased gradually (P<0.01).The protein of MMP-9 increased firstly and then was reduced (P<0.01),the peak was at the 24th hour.The protein of TIMP-1 was firstly transiently increased and then reduced (P<0.01), the minimum was at the 48th hour.Compared with the control group , the gene of IL-34 mRNA increased gradually (P<0.01), and the genes of CSF-1R mRNA, MMP-2 mRNA and MMP-9 mRNA increased firstly and then decreased ( P <0.01), which were peaked respectively at the 24th hour, the 24th hour, the 12th hour, respectively.Conclusion In the process of TGF-β1 inducing A549 cells transition,there is accompanied with the expression of IL-34/CSF-1R.