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@#Objective To develop and verify a method for detecting the activity of neutralizing antibodies in ELISA antibody positive serum of rats immunized with recombinant human interleukin-1 receptor antagonist(rhIL-1Ra). Methods The SD rats were subcutaneously immunized with 3,20 and 100 mg/kg rhIL-1Ra injection respectively,10 rats in each group,half male and half female,twice a day at an interval of at least 4 h between each dose for 13 consecutive weeks. The blood samples were collected from the jugular vein of rats during the administration period and the recovery period. The serum samples were isolated and detected for the antibody titers by ELISA,and the samples positive for rhIL-1Ra antibody were purified by Protein A chromatographic column. Based on,D10G4·1 cells biological activity assay,a method for the detection of neutralizing antibody activity was developed and verified for the specificity,sensitivity and precision. The neutralizing antibody activity of rhIL-1Ra antibody positive serum determined by ELISA was detected by using the developed method.Results With the increase of doses,the serum antibody titers of rats in various dose groups gradually increased,and there were still antibodies in the recovery period,and the titer was still high. Rabbit anti-rhIL-1Ra monoclonal antibody showed obvious neutralizing effect on rIL-1Ra,while rabbit anti-rIFN-2b monoclonal antibody had no dose-effect relationship with rIL-1Ra. The sensitivity of the method was 171. 93 μg/mL;The CVs of precision verification were not more than 20%. The positive antibody sera detected by ELISA all had neutralizing effect on rhIL-1Ra injection,which was consistent with the results detected by ELISA. Conclusion The method developed in this study has good specificity and high sensitivity in the detection of serum neutralizing antibody activity in rats immunized with rhIL-1Ra,which can be used to detect the serum neutralizing antibody activity of animals with rhIL-1Ra repeated administration.
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Interleukin-1 receptor associated kinase 4 (IRAK-4), acting as a serine threonine kinase, is considered as a key signal node for the transduction of IL-1R family and TLRs signal pathway. Studies have found that IRAK-4 has a hand in many signal pathways, involving the inflammatory response of human joints, intestines, liver and nervous system, as well as other autoimmune diseases. It is also one of the causes of drug resistance of some cancer cells. Therefore, IRAK-4 tends to be an effective therapeutic target for inflammatory diseases and cancer. The prospects for the development of drugs in this pathway is to develop novel IRAK-4 small molecule inhibitors and investigate their safety and effectiveness, enrich the clinical treatment of inflammatory and cancer diseases finally. This paper classified and summarized the latest research progress on small molecule inhibitors of IRAK-4 signaling pathway according to structures of the compounds, in order to provide assistances and references for the research and development of related drugs.
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Objective:To investigate the role and mechanism of miR-146b in the proliferation, metastasis and apoptosis of thyroid papillary carcinoma cells.Methods:qRT-PCR was used to detect the expression of miR-146b in thyroid papillary carcinoma cells (NPA, GLAG-66, ONCNO-DG1 and B-CPAP) and normal human thyroid cell line HTori3. After B-CPAP cells were transfected with miR-146b inhibitor, the inhibition efficiency was detected by qRT-PCR, the effect of miR-146b on PTC cells proliferation was detected by MTT assay, the effect of miR-146b on PTC cells invasion was studied by Transwell assay, and the effect of miR-146b on tumor cell apoptosis was detected by flow cytometry. SiRNA-IRAK1 was transfected into B-CPAP cell line. The cell proliferation rate, migration ability and apoptosis rate were detected by MTT, cell scratch test and flow cytometry respectively. The target gene of miR-146b, interleukin-1 associated receptor kinase 1 (IRAK1) , was predicted by bioinformatics software, and the regulatory effect of miR-146b on IRAK1 was verified by double fluorescein reporter gene experiment.Results:QRT-PCR showed that the expression of miR-146b in NPA87, KAT-5, FTC-133 and B-CPAP cell lines was significantly higher than that in normal cell HTori3, especially B-CPAP ( P<0.05) . MiR-146b inhibitor transfection could significantly reduce the expression level of miR-146b in B-CPAP cells ( P<0.01) . MTT results showed that miR-146b inhibitor could inhibit the proliferation of B-CPAP cells ( P<0.05) . Flow cytometry showed that miR-146b inhibitor could promote the apoptosis of B-CPAP cells ( P<0.05) . Transwell results showed that miR-146b inhibitor could reduce the invasive ability of B-CPAP cells ( P<0.05) . After transfection with siRNA-IRAK1, the proliferation rate of B-CPAP cells increased significantly (MTT test) , the migration ability increased (cell scratch test) , and the apoptosis rate decreased significantly (flow cytometry) ( P<0.05) . The results of double luciferase reporter gene showed that irak1 was the target gene of miR-146b, and miR-146b inhibitor could significantly up regulate the expression level of irak1 protein in B-CPAP cells. Conclusion:miR-146b may play a role in promoting the proliferation and metastasis and inhibiting cell apoptosis of PTC cells by inhibiting the downstream target protein IRAK1.
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Objective:To explore the expression level of interleukin-1 receptor-associated kinase-1 (IRAK1) in the peripheral blood of rheumatoid arthritis (RA) patients and analyze its relevance between disease activity and CD4 + T cell subsets. Methods:① The concentration of IRAK1 in the peripheral blood of 77 RA patients and 24 healthy controls were detected by enzyme linked immunosorbent assay (ELISA). ② The demo-graphic and clinical data of the RA group including disease activity score with 28 joints (DAS28), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), CD4 + T cell subsets in peripheral blood. ③Independent sample t test or Mann-Whitney U test were used to compare the differences between the two groups. Spearman rank correlation test and multiple linear regression were used to analyze the correlation between IRAK1 expression level and clinical data. Results:① The IRAK1 level of the peripheral blood of RA patients was significantly higher than in the normal controls ( P<0.001). ② Compared to normal controls, the peripheral blood of the RA group, the absolute numbers and proportion of regulatory T (Treg) cells were decreased ( P<0.001), the absolute numbers and proportion of helper T (Th) 17 and the ratio of Th17/Treg were increased. Moreover, the ratio of Th17/Treg was also increased. ③ With the increase of disease activity in RA patients, the expression of IRAK1 also increased. The expression of IRAK1 in the peripheral blood of RA group was positively correlated with ESR, number of joints involved and DAS28, and had statistically significant difference between the two groups ( r=0.23, P<0.05; r=0.24, P<0.05; r=0.27, P<0.05). Meanwhile, it was sign-ificantly negatively correlated with the percentage of Treg ( r=-0.27, P<0.05), and was significantly positively correlated with the ratio of Th17/Treg ( r=0.23, P<0.05) . However, there was no significant correlation with the ratio of Th1/Th2( P>0.05). Furthermore, multiple stepwise regression analysis showed that the expression of IRAK1 in the peripheral blood of RA group was positively correlated with ESR and the number of joints involved ( β=0.34, P=0.019; β=0.27, P=0.004), and it was inversely correlated with percentage of Treg ( β=-0.23, P=0.047, R2=0.219). Conclusion:IRAK1 expression in the peripheral blood of RA patients is up-regulated and correlated with disease activity. The decrease of Treg and the imbalance of Th17/Treg caused by high expression of IRAK1 may be one of the main factors for the occurrence and development of RA. Interfering the expression of IRAK1 may be a potential new target for RA treatment.
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Allogeneic hematopoietic stem cell transplantation (allo-HSCT) could effectively treat multiple hematological diseases. At present, with persistent improvement of transplantation techniques and rapid development of economy, more and more patients with hematological diseases are able to survive for a long time due to allo-HSCT treatment. Chronic ocular graft-versus-host disease (coGVHD) is the most common ocular complication after allo-HSCT, which is primarily manifested with refractory dry eye. In severe cases, it may cause imbalance of ocular surface homeostasis and limbal stem cell insufficiency, further leading to a series of complications that threaten the visual function and eye health, such as corneal perforation and symblepharon, etc. It is highly difficult to cure these symptoms. At present, relevant studies of clinical manifestations, diagnostic criteria, treatment specification and pathogenesis of coGVHD have been gradually deepened within the international community. However, related research and the establishment of clinical specification are still in the primary stage in China. In this article, research progress on clinical characteristics and related mechanisms of coGVHD was reviewed, aiming to deepen the understanding of this disease by ophthalmologists, especially specialists in corneal and ocular surface diseases, and provide novel ideas for subsequent in-depth research.
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Aim To explore the effect of dexmedetomidine (DEX) on acute lung injury (ALI) induced by lipopolysaccharide ( LPS) in rats and its mechanism. Methods The experiment included normal control group (Control) , DEX control group (DEX) , LPS-in-duced acute lung injury group (LPS) and LPS + DEX treatment group ( LPS + DEX ). Twenty-four hours after the successful modelling, the wet/dry weight ratio ( W/D) of the lung tissues of each group and the content of inflammatory cytokines IL-1 (3, TNF-a and IL-6 in the bronchoalveolar lavage fluid ( BALF) were measured; Hematoxylin-eosin (HE) staining was used to observe the pathological changes of lung tissues; im- munohistochemistry and Western blot were used to detect the expression of SIGIRR and NF-kB in lung tissues. Results Compared with control group, the levels of IL-1 (3, IL-6 and TNF-a in lung tissues of LPS group and BALF increased (P <0. 01). The lung tis sues showed obvious pathological damage of ALI, and the expression of SIGIRR was reduced ( P < 0. 01), the expression of phosphorylation activation of NF-kB increased (P <0. 01). Compared with LPS group, the W/D of lung tissues in LPS + DEX group and IL-1 [}, IL- in BALF 6 and TNF-a content were reduced (P < 0. 05 or P < 0. 01 ) . The pathological damage of lung tissues was significantly reduced, SIGIRR expression increased, and the activation of NF-kB phosphorylation decreased (P < 0. 01 ). Conclusions Dexmedetomidine can reduce SIGIRR degradation, inhibit the activation of NF-kB and reduce the inflammation, thereby reducing the acute lung injury induced by lipopolysaccharide in rats.
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Objective:To explore the expression and clinical significance of IL-1β and IL-1β receptor antagonist(IL-1ra)in persistent pulmonary hypertension of the newborn(PPHN)secondary to sepsis.Methods:The newborns with sepsis were enrolled in the Department of Neonatal Intensive Care Unit(NICU)of Xi′an Children′s Hospital from January 2018 to November 2020.The newborns with sepsis were divided into two groups: the newborns without PPHN( n=108)were the control group and the newborns with PPHN( n=44)were the experimental group.Clinical data, laboratory examination and bedside echocardiography of all the newborns were collected to analyze the differences between the two groups.The expression levels of IL-1β and IL-1ra in neonatal plasma of the two groups were detected by enzym-linked immunosorbination(ELISA), and their roles in neonatal sepsis with PPHN were further analyzed.The risk factors of neonatal sepsis with PPHN were analyzed by multivariate Logistic regression, and the early prediction value of the risk factors for neonatal sepsis with PPHN were evaluated by the receiver operating characteristic(ROC)curve. Results:There were no significant differences in gestational age[(39.11±0.55)w vs(38.85±0.72)w], birth weight[(3.30±0.49)kg vs(3.24±0.55)kg]and proportions of males[60(55.6%)vs 30(68.2%)]between the two groups( P>0.05). The right ventricular diameter[(9.57±0.35)mm], pulmonary artery pressure[(51.36±5.91)mmHg]and the level of N-terminal brain natriuretic peptide(NT-proBNP)[(25436.83±12343.18)ng/L)]significantly increased in the experimental group than those in the control group[(8.77±0.41)mm, (31.24±5.11)mmHg, (11267.09±4405.48)ng/L, respectively, P<0.05]. Before treatment, the expression levels of plasma IL-1β[(31.24±5.25)ng/L]and IL-1ra[(41.94±10.13)ng/L]in the experimental group were significantly higher than those in the control group[(18.27±4.47)ng/L, (21.47±8.76)ng/L, respectively, P<0.05]. The expression levels of plasma IL-1β[(10.46±3.17)ng/L]and IL-1ra[(10.58±2.94)ng/L]in the experimental group after treatment were significantly lower than those before treatment[(31.24±5.25)ng/L, (41.94±10.13)ng/L , respectively, P<0.05]. Multivariate Logistic regression analysis showed that IL-1β and NT-proBNP were the independent risk factors for neonatal sepsis with PPHN( P<0.05). ROC curve analysis showed that IL-1β and NT-proBNP had the good predictive value for the occurrence of neonatal sepsis with PPHN( P<0.05). IL-1β combined with NT-proBNP has the better predictive value for neonatal sepsis with PPHN. Conclusion:IL-1β combined with NT-proBNP have the high predictive value for PPHN of the newborns secondary to sepsis.
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Toll-like receptor 3 (TLR3) is a member of the TLR family, mediating the transcriptional induction of type I interferons (IFNs), proinflammatory cytokines, and chemokines, thereby collectively establishing an antiviral host response. Studies have shown that unlike other TLR family members, TLR3 is the only RNA sensor that is utterly dependent on the Toll-interleukin-1 receptor (TIR)-domain-containing adaptor-inducing IFN-β (TRIF). However, the details of how the TLR3-TRIF signaling pathway works in an antiviral response and how it is regulated are unclear. In this review, we focus on recent advances in understanding the antiviral mechanism of the TRIF pathway and describe the essential characteristics of TLR3 and its antiviral effects. Advancing our understanding of TLR3 may contribute to disease diagnosis and could foster the development of novel treatments for viral diseases.
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ABSTRACT BACKGROUND: Up to 5% of familial Mediterranean fever (FMF) cases are unresponsive to colchicine, through resistance, side effects and toxicity. Anakinra is an alternative treatment for FMF patients whose disease remains uncontrolled with colchicine. We aimed to evaluate anti-interleukin-1 treatment regarding clinical findings, laboratory parameters and quality of life (QoL) among FMF patients presenting resistance and toxicity towards colchicine. DESIGN AND SETTING: Descriptive observational study at the rheumatology clinic, Adnan Menderes University Medical School, Aydın, Turkey. METHODS: Among the patients included, age, sex, MEFV genotypes, acute-phase reactants, hepatic/renal function tests, average colchicine dose, disease duration, attack frequency, attack duration, disease severity, proteinuria, amyloidosis and QoL were evaluated. Colchicine resistance was defined as > 6 typical episodes/year or > 3 per 4-6 months. Kolmogorov-Smirnov, Friedman and two-way analysis of variance tests were used for statistical analyses. RESULTS: Between 2015 and 2017, 14 FMF patients receiving anakinra were enrolled. The mean colchicine dose was 1.7 ± 0.3 mg/day before use of anakinra. Ten patients were attack-free after treatment, while three showed reductions of at least 50% in attack frequency, attack duration and disease severity. Proteinuria levels in all patients with renal amyloidosis decreased after treatment. QoL among patients with renal amyloidosis differed significantly from QoL among non-amyloidosis patients. Mean visual analogue scale scores significantly improved in both groups after use of anakinra. CONCLUSIONS: Use of anakinra reduced attack frequency and proteinuria and acute-phase reactant levels, and improved QoL, with only a few uncomplicated side effects among colchicine-resistant or intolerant FMF patients. Injection-site reactions of severity insufficient to require discontinuation of treatment were seen.
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Humans , Male , Female , Adult , Middle Aged , Familial Mediterranean Fever/drug therapy , Quality of Life , Drug Resistance/drug effects , Colchicine/therapeutic use , Interleukin-1/antagonists & inhibitors , Antirheumatic Agents/therapeutic use , Interleukin 1 Receptor Antagonist Protein/therapeutic use , Familial Mediterranean Fever/physiopathology , Proteinuria/urine , Reference Values , Time Factors , Turkey , Severity of Illness Index , Blood Sedimentation , Reproducibility of Results , Retrospective Studies , Analysis of Variance , Treatment Outcome , Statistics, Nonparametric , Visual Analog Scale , Amyloidosis/physiopathology , Amyloidosis/drug therapy , Kidney Diseases/physiopathology , Kidney Diseases/drug therapyABSTRACT
Objective:To observe the Toll-like receptor 4(TLR4)and its negative regulating factorInterleukin-1 receptor-associated kinase-M(IRAK-M)in colonic mucosa of rats with experimental ulcerative colitis(UC), and to discuss the mechanism of the Chinese medicine Sishenwan. Method:The 90 Wistar rats were randomly divide into six groups, blank group, model group, sulfasalazine group(0.36 g·kg-1), Sishenwan low, medium and high-dose group(2.5, 5, 10 g·kg-1), 15 cases in each group. A rat model of UC was prepared by using a solution of trinitrobenzenesulfonic acid/ethano.The histopathological changes of colon were observed by hematoxylin-eosin (HE) staining. The contents of serum free triiodothyroid acid (FT3), serum free thyroxine (FT4), immunoglobulin (Ig) E and interleukin (IL)-2 were determined by radioimmunoassay. The activity of superoxide dismutase (SOD) in rat serum was determined by xanthine oxidation method. The activity of malondialdehyde (MDA) in serum of rats was determined by thiobarbituric acid (TBA) colorimetry. Result:Compared with blank group, intestinal mucosal injury score of rats in model group was significantly increased (PP3, FT4, IL-2 and SOD were significantly decreased (PPPPPP3, FT4, IL-2, and SOD contents were significantly increased (PPPPPConclusion:The unbalanced expressions of TLR4 and its negative regulating factor IRAK-M are connected with the pathogenesis of UC.Sishenwan can cure UC and control the expression of TLR4 and promote the expression of IRAK-M.
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OBJECTIVE: To study the effect of serum containing Jieduquyuziyin-prescription (JP) on signal pathway of interleukin-1 receptor-associated kinase 1 (IRAK1) in mononuclear macrophages of mice stimulated by lipopolysaccharide (LPS), and to explore the effect of Jieduquyuziyin-prescription on IRAK1 and NF-κB inflammatory signaling pathways, which providing a good theoretical support for its anti-inflammatory clinical medication. METHODS: In this study, mice mononuclear macrophages cultured in vitro were randomly divided into blank group, LPS group, JP serum group, blank serum group, LPS plus JP serum group, LPS plus blank serum group, IRAK1 inhibitor group, inhibitor plus LPS group, inhibitor plus JP serum group and inhibitor plus blank serum group. After intervention for 24 h, the activity of JP on macrophages was tested by CCK8 method. The IRAK1 expression in macrophages was tested by immunofluorescence chemical staining. The content of TNF-α in the supernatant of the cells was detected by ELISA. The mRNA expressions of IRAK1, NF-κB, TNF-α and IL-6 were detected by RT-PCR. The protein expressions of IRAK1, p-IRAK1 and NF-κB were detected by Western-blot. The LC-MS was used to detect the active ingredients in JP serum. RESULTS: The results show that 2.5% of JP serum is the optimal concentration. Jieduquyuziyin-prescription could down-regulate the expression of TNF-α and IL-6 and inhibit the expression of IRAK1 and activate NF-κB(P<0.05). Paeoniflorin and ferulic acid were detected in the JP serum. CONCLUSION: Jieduquyuziyin-prescription can inhibit the expression of IRAK1 and NF-κB in mouse monocyte-macrophage cells after LPS stimulation and provide a good theoretical support for its anti-inflammatory clinical medication.
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Acute gout subsides spontaneously within a certain period of time after onset, a process known as spontaneous remission of gouty inflammation. Its pathophysiological mechanism is related to the dynamic interaction of various components in the immune system, i. e. through the phagocytosis of autoimmune cells, the negative regulation of inflammatory mediators such as NLRP3 inflammasome, 1L-1R and Toll-like receptors, the recruitment of neutrophil extracellular traps at the site of crystal deposition and the formation of local chronic granuloma, to alleviate the necrotizing inflammation mediated by monosodium urate crystals. This review summarizes the molecular mechanism of spontaneous remission of gouty inflammation.
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OBJECTIVE: To observe the effect of electroacupuncture (EA) on neurological behavior and activity of Toll-like receptor 2 / nuclear factor kappa B (TLR2/NF-κB) signaling of the ischemic cerebral area in cerebral ischemia-reperfusion injury (CIRI) rats, so as to explore its mechanisms underlying improvement of CIRI. METHODS: A total of 120 male SD rats were randomly divided into blank control, sham operation, model, EA and EA＋NF-κB inhibitor (Pyrrolidine Dithiocarbamate Hydrochloride, PDTC, EA＋PDTC) groups which were further divided into 3, 7, 14 and 28 d subgroups (n＝6 in each subgroup). The CIRI model was established by occlusion of the middle cerebral artery for 90 min, followed by reperfusion. EA (1－20 Hz, 6 V) was applied to "Shuigou" (GV26), "Neiguan" (PC6), "Sanyinjiao" (SP6) and "Weizhong" (BL40) for 30 min, once a day for 28 days. For rats of the EA＋PDTC group, PDTC solution (120 mg/kg) was intraperitoneally injected on the 3rd day after successful modeling and before EA intervention. The neurological deficit severity (Zea Longa score) was assessed 3, 7, 14 and 28 days after modeling. The expression levels of TLR2, Interleukin-1 receptor-associated kinase (IRAK) and NF-κB mRNAs in the ischemic penumbra region of brain tissue were detected by real-time fluorescence quantitative PCR. RESULTS: Following modeling, the neurological deficit scores were significantly increased from the 3rd day on after CIRI (P<0.05), the expression levels of TLR2 mRNA on day 3, 7, 14 and 28, and IRAK mRNA on day 3 and 7, as well as NF-κB mRNA on day 3, 7 and 14 were significantly up-regulated in the model group relevant to the blank control group (P<0.05). After EA intervention, the neurological deficit scores were significantly decreased in the EA group on day 3, 7 and 28 and in the EA＋PDTC group on day 3, 7, 14 and 28 in comparison with those of the model group (P<0.05). In addition, the expression levels of TLR2 mRNA and NF-κB mRNA on day 3, 7 and 14 in the EA group, and on day 3, 7, 14 and 28 in the EA＋PDTC group, IRAK mRNA on day 3 in the EA and EA＋PDTC group were significantly down-regulated (P<0.05), but those of IRAK mRNA on day 14 and 28 in the EA group were significantly up-regulated in comparison with those of the model group (P<0.05). The effect of the EA＋PDTC was obviously superior to that of simple EA in down-regulating the expression of TLR2 (on day 28), and IRAK (on day 3, 14, 28), and NF-κB (on day 3, 7 and 14) (all P<0.05). CONCLUSION: EA stimulation can improve the symptoms of neurological deficits in CIRI rats, which may be related to its effect in suppressing the expression of TLR2, NF-κB and IRAK mRNAs of the ischemic cerebral tissue, i．e., down-regulating the activity of TLR2/NF-κB signaling.
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@#Interleukin-33(IL-33) is a new member of the interleukin-1 (IL-1) cytokine superfamily. It can activate mast cells, lymphocytes and macrophages to produce Th2 cytokines and plays a very important role in inflammation, infection, and autoimmune disease. The classical signal pathway of IL-33 includes the isotrimer of ST2 and interleukin-1 receptor accessory protein (IL-1 RAcP), which transduces signals into cells. The IL-33/ST2 signaling pathway affects bone metabolism by activating T and B lymphocytes. This article reviews the role of the IL-33/ST2 signaling pathway in bone metabolism. The results of a literature review showed that at present, scholars at home and abroad still dispute the role of IL-33 in bone metabolism. Some scholars believe that IL-33 can inhibit osteoclast formation, and IL-33 has been recently implicated in physiological bone remodeling. However, other scholars believe that IL-33 can promote osteoclast formation and differentiation, which leads to bone absorption. IL-33 and its signaling pathway are involved in bone metabolism of alveolar bone in periodontitis and periapical periodontitis. The specific mechanism remains unclear, and further studies are warranted.
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Objective The study aimed to investigate the clinical value of serum soluble ST2 (sST2) and galectin-3(gal-3) in dilated cardiomyopathy(DCM) patients associated with heart failure.Methods 60 DCM patients with heart failure were collected in Anzhen Hospital Department of Cardiology from June to December 2017 as researching group.52 coronary heart disease(CHD) patients with heart failure were as the controls,also,70 healthy controls were analyzed by cross-sectional investigation.In DCM group,18 patients (male/female:11/7) were enrolled in the NYHA(New York Heart Association) cardiac function Ⅰ-Ⅱ group and 42 patients (male/female:32/10) were enrolled in the NYHA cardiac function Ⅲ-Ⅳ group.sST2 was detected by enzyme-linked immunoassay;gal-3 was detected by chemiluminescence immunoassay.One-Way ANOVA,non-parametric test,Spearman linear correlation analysis and other statistical methods were used in our study.Results Compared with healthy controls,the serum level of sST2 in DCM group increased markedly [21.23(14.59,25.29) ng/ml vs 14.59(12.78,17.16) ng/ml,standardized H=-4.645,adjusted P＜0.001],and significantly higher serum level of gal-3 [13.50(10.48,19.68) μg/L vs 10.05(8.25,12.80) μg/L,standardized H=4.266,adjusted P＜0.001].Compared with CHD group,there was a significant increase [13.50(10.48,19.68) μg/L vs 11.00(8.65,13.58) μg/L,standardized H=-2.715,adjusted P＝).020] in the expression of serum gal-3 in DCM group.The serum level of sST2 in cardiac function Ⅲ-Ⅳ group was obviously higher than that in the Ⅰ-Ⅱ group [22.54(16.82,26.78) ng/ml vs 12.24(9.22,17.79) ng/ml,Z=2.613,P＝0.009].Spearman correlation analysis showed a negative correlation between sST2 and left ventricular ejection fraction(LVFE) (r=-0.647,P＜0.01),but a positive correlation between left ventricular end-diastolic diameter (LVEDD) and left ventricular end-systolic dimension(LVESD) (r=0.393,r=0.462,all P＜0.01).The correlation of sST2 and B-type natriuretic peptide(BNP)/gal-3 (r=0.320,r=0.331,all P＜0.05),gal-3 and high-sensitivity troponinI(hs-TnI)/BNP(r=406,r=0.401,all P≤0.01) presented the same.There was a inverse correlation between gal-3 and estimated glomerular filtration rate(eGFR)(r=-0.326,P＜0.05).ROC analysis showed that the cutoff of sST2 was 21.00 ng/ml,with 51.9％ sensitivity and 95.7％ specificity (AUC=0.745,95％ CI:0.652-0.838,P＜0.001),and the cutoff of gal-3 was 13.15 μg/L,with 53.7％ sensitivity and 85.7％ specificity (AUC=0.720,95％ CI.0.627-0.812,P＜0.001).Combined detection of BNP/hs-TnI may remarkably improved the sensitivity (up to 95.3％).Conclusions The serum levels of sST2 and gal-3 were significantly increased in dilated cardiomyopathy patients with heart failure.It showed a strong relation between sST2 and the deterioration of cardiac function,and a certain correlation with left ventricular remodeling.Although the diagnostic value of DCM was lower than that of BNP,sST2 may not be affected by age,body mass index and the damage to kidney function.Combined diagnosis could dramatically improve the diagnostic efficacy.
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BACKGROUND: The liver is an organ with remarkable regenerative capacity; however, once chronic fibrosis occurs, liver failure follows, with high mortality and morbidity rates. Continuous exposure to proinflammatory stimuli exaggerates the pathological process of liver failure; therefore, immune modulation is a potential strategy to treat liver fibrosis. Mesenchymal stem cells (MSCs) with tissue regenerative and immunomodulatory potential may support the development of therapeutics for liver fibrosis. METHODS: Here, we induced hepatic injury in mice by injecting carbon tetrachloride (CCl₄) and investigated the therapeutic potential of conditionedmedium from tonsil-derivedMSCs (T-MSCCM). In parallel, we used recombinant human IL-1Ra,which, as we have previously shown, is secreted exclusively from T-MSCs and resolves the fibrogenic activation of myoblasts. Hepatic inflammation and fibrosis were determined by histological analyses using H&E and Picro-Sirius Red staining. RESULTS: The results demonstrated that T-MSC CM treatment significantly reduced inflammation as well as fibrosis in the CCl₄-injured mouse liver. IL-1Ra injection showed effects similar to T-MSC CM treatment, suggesting that T-MSC CM may exert anti-inflammatory and anti-fibrotic effects via the endogenous production of IL-1Ra. The expression of genes involved in fibrosis was evaluated, and the results showed significant induction of alpha-1 type I collagen, transforming growth factor beta, and tissue inhibitor of metalloproteases 1 upon CCl₄ injection, whereas treatment with T-MSC CM or IL-1Ra downregulated their expression. CONCLUSION: Taken together, these data support the therapeutic potential of T-MSC CM and/or IL-1Ra for the alleviation of liver fibrosis, as well as in treating diseases involving organ fibrosis.
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Animals , Humans , Mice , Carbon Tetrachloride , Collagen Type I , Culture Media, Conditioned , Fibrosis , Inflammation , Interleukin 1 Receptor Antagonist Protein , Liver Cirrhosis , Liver Failure , Liver , Mesenchymal Stem Cells , Metalloproteases , Mortality , Myoblasts , Transforming Growth Factor betaABSTRACT
Background: Vitiligo is a multifactorial, polygenic, autoimmune skin disorder caused by selective destruction of melanocytes. Interleukin 1 receptor antagonist intron 2 polymorphism was found to be associated with various autoimmune disorders. Aims: We aimed to investigate the association of interleukin 1 receptor antagonist intron 2 variable number of tandem repeats polymorphism (rs2234663) with vitiligo to assess interleukin 1 receptor antagonist transcript levels and to perform possible genotype–phenotype correlation. Methods: Three hundred and seven vitiligo patients and 316 controls were enrolled in the study, genotyping of interleukin 1 receptor antagonist rs2234663 was performed by polymerase chain reaction, and relative gene expression of interleukin 1 receptor antagonist was carried out in peripheral blood mononuclear cells from patients (n = 36) and controls (n = 36) by real-time-PCR. Results: A significant difference was observed in the frequency of interleukin 1 receptor antagonist *A (1/2) genotype among patients with active and stable vitiligo (P = 0.0172). Interleukin 1 receptor antagonist*A (2/2) genotype and allele frequencies were significantly different between SV patients and controls (P = 0.0246 and P = 0.0046, respectively). Significant difference was also observed for interleukin 1 receptor antagonist*A2 (allele) in active and stable vitiligo patients (P = 0.0060). However, other comparisons did not show any significant difference in genotype and allele frequencies. Moreover, interleukin 1 receptor antagonist*A (3/2) genotype was observed only in patients whereas interleukin 1 receptor antagonist*A (5/2) was observed only in controls. Gene expression analysis showed no significant difference in interleukin 1 receptor antagonist transcript levels in patients compared to controls (P = 0.5962). Interestingly, genotype–phenotype correlation analysis revealed that individuals with IL1RN*A (2/2) exhibited higher interleukin 1 receptor antagonist expression compared to other major genotypes interleukin 1 receptor antagonist*A (1/2) (P = 0.01) and interleukin 1 receptor antagonist*A (1/1) (P = 0.03). Limitations: More case-control studies on interleukin 1 receptor antagonist rs2234663 polymorphism and gene expression from different ethnic populations are required to explore the impact of interleukin 1 receptor antagonist in vitiligo susceptibility. Conclusion: Interleukin 1 receptor antagonist*A2 might be a risk factor for progressive vitiligo.
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Objective • To amplify the interferon regulator factor 3 (IRF3) short hairpin RNA (shRNA) virus and investigate the effect of the virus on the nuclear expression of Irak1bp1 protein in lipopolysaccharide (LPS)-stimulated Raw 264.7 cells. Methods • Adenovirus was amplified in HEK293T cells and the virus titer was detected by TCID 50 assay. The Raw 264.7 cells were randomly divided into four groups including adenovirus (-) LPS (-) group, adenovirus (-) LPS (+) group, adenovirus (+) LPS (-) group and adenovirus (+) LPS (+) group. The expression of intracellular IRF3 mRNA was detected by real-time PCR, and the nuclear expression of IRF3 and Irak1bp1 protein were detected by Western blotting. Results • The titer of adenovirus was 2.2×1011 PFU/mL and the best MOI was 300. The expression of IRF3 mRNA and nuclear IRF3 protein in LPS-stimulated Raw 264.7 cells were significantly higher than those of the control group. The cellular constitutive expression of IRF3 at mRNA level and the LPS-induced expression of IRF3 were significantly inhibited after transfection of Raw 264.7 cells with adenovirus strains carrying IRF3 shRNA. However, the nuclear constitutive expression of IRF3 protein was not affected by IRF3 shRNA in the unstimulated state. The expression of nuclear Irak1bp1 protein was significantly higher than that of the control group. The nuclear constitutive expression and the LPS-induced expression of Irak1bp1 protein were not affected by IRF3 shRNA. Conclusion • Transfection of LPS-stimulated Raw 264.7 cells with adenovirus strains carrying IRF3 shRNA could effectively inhibit the expression of IRF3, but not affect the nuclear expression of Irak1bp1 protein.
ABSTRACT
Inflammation response is an important pathological process of neuronal cell death after stroke.Interleukin (IL)-33/ST2 system is involved in the inflammatory response process of ischemic stroke and has been proved to be a protective factor.This article reviews the role and mechanism of IL-33/ST2 system in the regulation of immune inflammation after ischemic stroke.
ABSTRACT
Chronic urticaria may often be associated with interleukin (IL)-1-mediated autoinflammatory disease, which should be suspected if systemic inflammation signs are present. Here, we report a case of Schnitzler's syndrome without monoclonal gammopathy treated successfully with the IL-1 receptor antagonist anakinra. A 69-year-old man suffered from a pruritic urticarial rash for 12 years. It became aggravated episodically and was accompanied by high fever, arthralgia, leukocytosis, and an elevated C-reactive protein and erythrocyte sedimentation rate. The episodes each lasted for over one week. Neutrophilic and eosinophilic inflammation was found on skin biopsy. However, serum and urine electrophoresis showed no evidence of monoclonal gammopathy. The cutaneous lesions were unresponsive to various kinds of anti-histamines, systemic glucocorticoids, colchicine, cyclosporine, dapsone, and methotrexate, which were administered over a span of 3 years immediately preceding successful treatment. A dramatic response, however, was observed after a daily administration of anakinra. This observation suggests that the correct diagnosis of this case is Schnitzler's syndrome without monoclonal gammopathy. For an adult patient with refractory chronic urticaria and systemic inflammation, Schnitzler's syndrome could be considered as a possible differential diagnosis. Although the typical form of Schnitzler's syndrome exhibits the presence of monoclonal gammopathy as a diagnostic criterion, monoclonal gammopathy may be absent in an atypical form. In such a situation, an IL-1 antagonist should be effective for the management of chronic urticaria.