Amounts Of IL-1β Research Articles

25-hydroxycholesterol promotes brain cytokine production and leukocyte infiltration in a mouse model of lipopolysaccharide-induced neuroinflammation

Neuroinflammation has been implicated in the pathogenesis of several neurologic and psychiatric disorders. Microglia are key drivers of neuroinflammation and, in response to different inflammatory stimuli, overexpress a proinflammatory signature of genes. Among these, Ch25h is a gene overexpressed in brain tissue from Alzheimer’s disease as well as various mouse models of neuroinflammation. Ch25h encodes cholesterol 25-hydroxylase, an enzyme upregulated in activated microglia under conditions of neuroinflammation, that hydroxylates cholesterol to form 25-hydroxycholesterol (25HC). 25HC can be further metabolized to 7α,25-dihydroxycholesterol, which is a potent chemoattractant of leukocytes. We have previously shown that 25HC increases the production and secretion of the proinflammatory cytokine, IL-1β, by primary mouse microglia treated with lipopolysaccharide (LPS). In the present study, wildtype (WT) and Ch25h-knockout (KO) mice were peripherally administered LPS to induce an inflammatory state in the brain. In LPS-treated WT mice, Ch25h expression and 25HC levels increased in the brain relative to vehicle-treated WT mice. Among LPS-treated WT mice, females produced significantly higher levels of 25HC and showed transcriptomic changes reflecting higher levels of cytokine production and leukocyte migration than WT male mice. However, females were similar to males among LPS-treated KO mice. Ch25h-deficiency coincided with decreased microglial activation in response to systemic LPS. Proinflammatory cytokine production and intra-parenchymal infiltration of leukocytes were significantly lower in KO compared to WT mice. Amounts of IL-1β and IL-6 in the brain strongly correlated with 25HC levels. Our results suggest a proinflammatory role for 25HC in the brain following peripheral administration of LPS.

Neisseria meningitidis activates pyroptotic pathways in a mouse model of meningitis: role of a two-partner secretion system.

There is evidence that in infected cells in vitro the meningococcal HrpA/HrpB two-partner secretion system (TPS) mediates the exit of bacteria from the internalization vacuole and the docking of bacteria to the dynein motor resulting in the induction of pyroptosis. In this study we set out to study the role of the HrpA/HrpB TPS in establishing meningitis and activating pyroptotic pathways in an animal model of meningitis using a reference serogroup C meningococcal strain, 93/4286, and an isogenic hrpB knockout mutant, 93/4286ΩhrpB. Survival experiments confirmed the role of HrpA/HrpB TPS in the invasive meningococcal disease. In fact, the ability of the hrpB mutant to replicate in brain and spread systemically was impaired in mice infected with hrpB mutant. Furthermore, western blot analysis of brain samples during the infection demonstrated that: i. N. meningitidis activated canonical and non-canonical inflammasome pyroptosis pathways in the mouse brain; ii. the activation of caspase-11, caspase-1, and gasdermin-D was markedly reduced in the hrpB mutant; iii. the increase in the amount of IL-1β and IL-18, which are an important end point of pyroptosis, occurs in the brains of mice infected with the wild-type strain 93/4286 and is strongly reduced in those infected with 93/4286ΩhrpB. In particular, the activation of caspase 11, which is triggered by cytosolic lipopolysaccharide, indicates that during meningococcal infection pyroptosis is induced by intracellular infection after the exit of the bacteria from the internalizing vacuole, a process that is hindered in the hrpB mutant. Overall, these results confirm, in an animal model, that the HrpA/HrpB TPS plays a role in the induction of pyroptosis and suggest a pivotal involvement of pyroptosis in invasive meningococcal disease, paving the way for the use of pyroptosis inhibitors in the adjuvant therapy of the disease.

Influence of locally delivered doxycycline on the clinical and molecular inflammatory status of intrabony defects prior to periodontal regeneration: A double-blind randomized controlled trial.

To test the effect of locally delivered doxycycline (DOX) administered 2 weeks prior to minimally invasive periodontal regeneration in terms of presurgical inflammatory status and cytokine expression profile in the gingival crevicular fluid (GCF). Secondary aim was to assess the early wound healing index (EHI) at 2 weeks after surgery. It is hypothesized that healing after periodontal regeneration is dependent on preoperative soft tissue condition, and that local antibiotics may improve the site-specific inflammatory status at short time. Sites associated with periodontal intrabony defects requiring regenerative surgery and showing bleeding on probing (BoP) were included. At T0, experimental sites were randomly treated with subgingival instrumentation with or without topic DOX application. After 2 weeks (T1), defects were approached by means of minimally invasive surgical technique. GCF was sampled at both T0 and T1 for inflammatory biomarker analysis. Two weeks after surgery, the EHI was evaluated (T2). Forty-four patients were included. At T1, the number of BoP+ sites was statistically significantly less in the test group (27.3% vs. 72.7%; p < .01). The total amount of interleukin (IL)-1β (p < .001), matrix-metalloproteinases (MMP)-8 (p < .001), and MMP-9 (p = .010) in the GCF significantly decreased in the test group at T1, with relevant differences compared to controls. At T2, the EHI had an average value of 1.45 ± 0.86 in the test group while in the control, it was 2.31 ± 1.43 (p = .027). A statistically significantly positive correlation was observed between the amount of IL-1β and MMP-9 and EHI scores. Within the limitations of this study, sites treated with DOX showed improved clinical and molecular inflammatory parameters before surgery, as well as soft tissue healing 2 weeks after surgery.

Evaluation of Interleukin-1β and Tumor Necrosis Factor Alpha Concentration in a Sample of Endodontic Patients in Hillah City, Iraq

Abstract Background: Endodontic illness, one of the most common chronic infectious diseases in the world, is brought on by an infected and inflamed dental pulp, which results in bone destruction around the tooth root, excruciating pain, and tooth loss. Objectives: This study aimed to determine levels of pro-inflammatory (interleukin-1β [IL-1β] and tumor necrosis factor alpha [TNF-α]) by enzyme-linked immune sorbent assay in the saliva of patients with endodontic infection against the gender and age groups. Materials and Methods: From November 2021 to March 2022, 60 patients (n = 60) were referred to specialized dentistry centers and private clinics in Hillah City, Iraq, with 32 (53.33%) females and 28 (46.67%) males, and 20 (n = 20) healthy adults serving as study controls, all of whom were between the ages of 20 and 50. Results: There were no major differences between girls and males in both groups, according to the findings of this study. In comparison to the control group, however, patients showed higher significant differences in measuring (IL-1β) and (TNF-α) concentrations (180.8), (119.44) (P ≤ 0.05), respectively. Furthermore, the amounts of IL-1β and TNF-α in female saliva were substantially greater than in male saliva. Conclusion: Pro-inflammatory IL-1β and TNF-α were shown to be higher in patients with primary endodontic disease than in normal subjects.

Local cytokine levels as prognostic factors for early relapse of non-muscle-invasive bladder carcinoma

The aim of our study is to assess the local cytokine levels as prognostic factors for early relapse in NMIBC patients. 75 patients with NMIBC were enrolled in the study: 51 with primary NMIBC and 24 with initially recurrent NMIBC, LG and HG tumors were diagnosed in each group. Patients with primary NMIBC were monitored during 9 months after treatment: TURB and chemotherapy (No. 6). During TURB samples of tumors were taken, supernatants were obtained and tissue cytokine levels were measured (IL-1β, IL-6, IL-10, IL-18, TNFα, IFNγ, IL-8) by ELISA test. The results showed that in patients with primary NMIBC early relapses were diagnosed in 15 (46.8%) of LG tumors and in 11 (45%) of HG tumors matching that there was no difference depending upon tumor grade. In initially recurrent tumors of both LG and HG NMIBC the amounts of cytokines were maximal: in LG tumors they exceeded the primary ones from 7.1 (IFNγ) to 300 (IL-6) while in HG - from 2.0 (IL-10) to 9.7 (IL-6). The amounts of IL-1β, IL-6, IL-10, IFNγ, IL-8 were higher in those LG primary tumors which relapsed in 6-9 months compared to the ones which didn't, though their levels were much lower than in initially manifested relapse (from 2.6 times for IFNy to 150 times for IL-6). A similar trend, though not for all the same cytokines, was observed in HG tumors: tissue levels of IL-6, IL-10, IL-18 and TNFα were higher in tumors which relapsed in 6-9 months after treatment. The increase of 2 cytokines' levels were common for both LG and HG tumors (IL-6 and IL-10). This finding might be considered as a new prognostic factor of the early relapse. We conclude that relapse of LG and HG NMIBC is related to some immune mechanisms, namely to local hyperproduction of cytokines, especially IL-6 and IL-10, though IL-1β, IL-8, IFNγ could have an impact on LG and IL-18, TNFα — on HG tumors. Taking into account common signaling pathways of IL-6 and IL-10 like JAK/STAT, these transcription factors might be potential targets for new effective approaches to treatment.

Analysis of target organs of Houttuynia cordata: A study on the anti-inflammatory effect of upper respiratory system

Ethnopharmacological relevanceHouttuynia cordata Thunb. (HC) is a traditional anti-pyretic herb that is classified as the lung meridian in traditional Chinese medicine. However, no articles have explored the main organs responsible for the anti-inflammatory activities of HC.Aim of the study: The aim of the study was to investigate the meridian tropism theory of HC in lipopolysaccharide (LPS)-induced pyretic mice, as well as to identify the underlying mechanisms. Materials and methodsTransgenic mice carrying the luciferase gene driven by nuclear factor-κB (NF-κB) were intraperitoneally injected with LPS and orally administered standardized concentrated HC aqueous extract. The phytochemicals present in the HC extract were analyzed using high-performance liquid chromatography. In vivo and ex vivo luminescent imaging from transgenic mice was used to investigate the meridian tropism theory and anti-inflammatory effects of HC. Microarray analysis of gene expression patterns was used to elucidate the therapeutic mechanisms of HC. ResultsHC extract was found to contain phenolic acids, such as protocatechuic acid (4.52%) and chlorogenic acid (8.12%), as well as flavonoids like rutin (2.05%) and quercitrin (7.73%). The bioluminescent intensities induced by LPS in the heart, liver, respiratory system, and kidney were significantly suppressed by HC, while the maximal decrease (about 90% reduction) of induced luminescent intensity was observed in the upper respiratory tract. These data suggested that upper respiratory system might be the target for HC anti-inflammatory abilities. HC affected the processes involved in innate immunity, such as chemokine-mediated signaling pathway, inflammatory response, chemotaxis, neutrophil chemotaxis, and cellular response to interleukin-1 (IL-1). Moreover, HC significantly reduced the proportions of p65-stained cells and the amount of IL-1β in trachea tissues. ConclusionBioluminescent imaging coupled with gene expression profile was used to demonstrate the organ selectivity, anti-inflammatory effects, and therapeutic mechanisms of HC. Our data demonstrated for the first time that HC displayed lung meridian-guiding effects and exhibited great anti-inflammatory potential in the upper respiratory tract. The NF-κB and IL-1β pathways were associated with the anti-inflammatory mechanism of HC against LPS-provoked airway inflammation. Moreover, chlorogenic acid and quercitrin might be involved in the anti-inflammatory properties of HC.

Gene expression of Toll-like receptors, cytokines and a nuclear factor and cytokine secretion in DH82 canine macrophage cells infected with Brucella canis

Canine brucellosis caused by Brucella canis infection occurs mainly in dogs, and is a zoonotic disease that also has the possibility of infection in humans. Many studies have been conducted to understand the immunopathological mechanism of B. canis infection. However, the precise immune mechanism remains to be elucidated because compared to other Brucella spp., B. canis has different immune evasion mechanisms. In this study, gene expression levels of Toll-like receptors (TLRs) and TLR-associated molecules and cytokine production were analyzed to figure out the roles of immune-related host factors in B. canis infection. Time-dependent gene expression of TLRs (1−10) and TLR-related molecules (TNF-α, IL-5, IL-23, CCL4, CD40 and NFκ-B) and release of Th1, Th2 and Th17-related cytokines (IFN-γ, IL-1β, IL-4, IL-6, IL-10 and IL-17A) were investigated in DH82 canine macrophages infected with B. canis. Time-dependent induction of TLRs 3, 7 and 8 was observed, and TLR 7 had the highest expression level (p <0.05). The expression levels of all TLR-related genes were significantly increased after infection. In particular, the expression of the CCL4 and IL-23 genes was highly induced. The amounts of IL-1β, IL-6 and IL-10 were significantly increased by B. canis infection, but the amounts of IL-4 and IL-17A were not. The production of IL-1β and IL-6 was the highest at 24 hr after B. canis infection (p <0.05). This study demonstrates that TLRs 3, 7 and 8 are prominent sites of to immune response induction with the production of related cytokines and a nuclear factor in DH82 cells infected with B. canis. These results suggest a sequential immune mechanism of B. canis infection, involving TLRs, cytokines and their associated factors.

Evaluation of IL-1β and IL-6 Expression following EBNA-1 and BRLF-1 Peptide Treatment in Epstein-Barr Virus-Positive Multiple Sclerosis Patients

Introduction: Epstein-Barr virus (EBV/HHV-4) has been implicated in the pathogenesis of multiple sclerosis (MS). This study was conducted to investigate the levels of pro-inflammatory cytokines IL-1β and IL-6 in healthy EBV carriers and MS patients with prior EBV infection in response to treatment with EBV nuclear antigen 1 (EBNA-1) and replication and transcription activator (BRLF-1/Rta) peptide antigens in whole blood cell culture to assess the cytokine expression across all cells in the peripheral blood. Methods: Isolated whole blood cells from the included participants were incubated at a concentration of 10⁶ cells/mL with BRLF-1 or EBNA-1. The amount of IL-1β and IL-6 transcripts were measured with quantitative RT-PCR at day 3 after incubation. MTT assay was conducted to examine cytotoxicity of the peptides and their effect on cell viability. Changes in cytokine expression and cell viability were analyzed using one-way and two-way ANOVA, respectively. Results: Ten MS patients and ten healthy donors were enrolled in the study. Treatment with the peptide antigens resulted in increased cytokines expression in both MS patients and healthy subjects. Furthermore, IL-1β levels were higher in MS patients compared to healthy EBV carriers. MTT assay revealed no significant difference in cell viability between the two groups. Discussion: The higher levels of IL-1β in response to EBV antigens in MS patients may reflect the host neuroinflammatory environment and support the notion that immune response against EBV has a role as an aggravating factor in the progression of MS by contributing to the neuroinflammatory cascade.

A Comparative Evaluation of the Measurement of Interleukin-1β as Salivary Biomarker in Periodontitis Patients with and without Diabetes Mellitus

Introduction: Periodontitis is characterized by inflammatory destruction of connective tissue, loss of periodontal attachment, and resorption of the alveolar bone. Because of its activity as an inflammatory mediator as well as a modulator of the extracellular matrix and bone, the pro-inflammatory cytokine interleukin (IL)-1β.&#x0D; Aim: The study aimed to evaluate Interleukin-1β as a salivary biomarker in periodontitis patients with and without diabetes mellitus.&#x0D; Materials and methods: The study included 30 patients, 15 males and 15 females, ranging in age from 30 to 60 years old. Group A: Clinically healthy, Group B: Periodontitis with diabetes mellitus, and Group C: Periodontitis without diabetes mellitus were the three groups of patients under this investigation. Expectoration into sterile bulbs was used to collect whole unstimulated saliva from all patients. Following that, collected samples are sent to the laboratory, where the amount of IL-1β, as a periodontal disease marker was determined using the ELISA sandwich technique. The data were statistically analysed using One-Way ANOVA. The Newman-Keuls multiple comparison test was used to test the significance at the levels of p&lt;0.05.&#x0D; Results: IL-1β level was found to be significantly higher (p&lt;0.05) in periodontitis with diabetes mellitus (75 ±7 pg/L) when compared with patients with periodontitis only (45.06±7 pg/L) compared with healthy controls (36.37±5.6 pg/L).&#x0D; Conclusion: From the study, it was concluded that IL-1β level was significantly increased with periodontitis patients, and patients with periodontitis and diabetes mellitus, when compared to patients with clinically healthy group. In summary, diabetes mellitus and periodontal diseases are closely associated with each other. Salivary IL-1β level can be used as a biomarker in early diagnosis of chronic periodontitis in patients with diabetes mellitus.

Relationship Between Curative Effect and Serum Inflammatory Factors Level in Male Patients With First-Episode Schizophrenia Treated With Olanzapine.

Background: A growing body of evidence shows that immune system disorders are one of the important etiological factors of schizophrenia. Inflammatory cytokines play a very critical role in the pathogenesis and treatment of schizophrenia. However, in the actual clinical practice, there is still a lack of confirmed biological indicators that can be used to evaluate the therapeutic effect of antipsychotics.Methods: In this study, 82 male patients with first-episode schizophrenia and 30 healthy controls were included. The Positive and Negative Syndrome Scale (PANSS) scores were evaluated, and the serum levels of high-sensitivity C-reactive protein (hs-CRP), interleukin 1β (IL-1β), interleukin 6 (IL-6), interleukin 17 (IL-17), and transforming growth factor β1 (TGF-β1) were detected, both at baseline and 4 weeks later. The patients were divided into two groups, the effective group and the ineffective group, according to the reduction rate of PANSS.Results: In the case group, the levels of hs-CRP were significantly elevated (p = 0.00), whereas IL-1β, IL-6, and IL-17 were significantly reduced as compared to the baseline (p = 0.01, 0.02, and 0.00, respectively). Importantly, the baseline levels of the five inflammatory factors were significantly higher in the case group as compared to the control group (p = 0.00, 0.00, 0.00, 0.00, and 0.00, respectively). Post-treatment, the serum levels for IL-1β, IL-6, and IL-17 were significantly higher in the effective group than in the ineffective group (p = 0.00, 0.00, and 0.01, respectively). For every increase in the amount of IL-1β, the risk of ineffectiveness increased by 7% (OR = 0.93 [0.86–1.00]; p = 0.04), whereas for every increase in the amount of IL-17, the risk of ineffectiveness increased by 5% (OR = 0.95 [0.90–0.99]; p = 0.03).Conclusion: The results of the study showed that the levels of inflammatory factors in patients with different therapeutic effects were different, and the changes in the amounts of IL-1β and IL-17 acted as predictors of poor efficacy.

The frequency of interleukin-1β-producing monocytes is significantly associated with varicella-zoster responses of nursing home residents.

Previous studies have demonstrated that the status of the T cell compartment and inflammation-related factors are associated with the immunogenicity of the varicella-zoster virus (VZV) vaccine in older adults; however, little is known about the roles of other immune cell subsets known to influence the generation and maintenance of immunological memory. Responses to a live-attenuated VZV vaccine were studied in relation to peripheral blood mononuclear cell (PBMC) composition and function in a sample of 30 nursing home residents (aged 80-99 years). Interferon-gamma enzyme-linked immunospot (ELISPOT) was used to measure VZV responses at baseline and 6weeks following vaccination, and associations were sought with the frequencies of monocytes and T, B and natural killer (NK) cells and the production and secretion of cytokines following their ex-vivo stimulation with different agents. While only the frequency of interleukin (IL)-6+ CD14+ monocytes was inversely associated with post-vaccination VZV response, amounts of IL-1β, IL-10, IL-17A and tumour necrosis factor (TNF) secreted by PBMCs and the frequency of IL-1β+ CD14+ monocytes was positively correlated with pre-vaccination VZV response. Furthermore, both bivariate correlation and causal mediation analyses supported the notion that IL-1β+ CD14+ monocytes were significant mediators of the associations between IL-1β and TNF secretion by PBMCs and pre-vaccination VZV responses. Our findings implicate a strong cytokine response mediated by inflammatory IL-1β+ monocytes in coordinating responses of long-lived VZV-reactive memory T cells, but with an opposing effect of IL-6+ CD14+ monocytes. Whether monocyte status promotes or inhibits the induction and/or maintenance of these memory T cells later in life has yet to be determined.

Decoy Receptor 3 Inhibits Monosodium Urate-Induced NLRP3 Inflammasome Activation via Reduction of Reactive Oxygen Species Production and Lysosomal Rupture.

Gout is a common inflammatory arthritis caused by the deposition of monosodium urate (MSU) crystals in the joints. This activates the macrophages into a proinflammatory state by inducing NLRP3-dependent interleukin-1β (IL-1β) secretion, resulting in neutrophil recruitment. Soluble decoy receptor 3 (DcR3) is an immune modulator and can exert biological functions via decoy and non-decoy actions. Previously, we showed that DcR3 suppresses lipopolysaccharides (LPS)- and virus-induced inflammatory responses in the macrophages and promotes the macrophages into the M2 phenotype. In this study, we clarified the actions of DcR3 and its non-decoy action motif heparin sulfate proteoglycan (HSPG) binding domain (HBD) in the MSU crystal-induced NLRP3 inflammasome activation in the macrophages and in mice. In bone marrow-derived macrophages, THP-1 and U937 cells, we found that the MSU crystal-induced secretion of IL-1β and activation of NLRP3 were suppressed by both DcR3.Fc and HBD.Fc. The suppression of the MSU-induced NLRP3 inflammasome activation is accompanied by the inhibition of lysosomal rupture, mitochondrial production of the reactive oxygen species (ROS), expression of cathepsins, and activity of cathepsin B, without affecting the crystal uptake and the expression of NLRP3 or pro-IL-1β. In the air pouch mice model of gout, MSU induced less amounts of IL-1β and chemokines secretion, an increased M2/M1 macrophage ratio, and a reduction of neutrophil recruitment in DcR3-transgenic mice, which expresses DcR3 in myeloid cells. Similarly, the mice intravenously treated with DcR3.Fc or HBD.Fc displayed less inflammation response. These findings indicate that HBD of DcR3 can reduce MSU crystal-induced NLRP3 inflammasome activation via modulation of mitochondrial and lysosomal functions. Therefore, we, for the first time, demonstrate a new therapeutic potential of DcR3 for the treatment of gout.

Activation of Nurr1 with Amodiaquine Protected Neuron and Alleviated Neuroinflammation after Subarachnoid Hemorrhage in Rats

Background. Nurr1, a member of the nuclear receptor 4A family (NR4A), played a role in neuron protection, anti‐inflammation, and antioxidative stress in multidiseases. We explored the role of Nurr1 on subarachnoid hemorrhage (SAH) progression and investigated the feasibility of its agonist (amodiaquine, AQ) as a treatment for SAH. Methods. SAH rat models were constructed by the endovascular perforation technique. AQ was administered intraperitoneally at 2 hours after SAH induction. SAH grade, mortality, weight loss, neurological performance tests, brain water content, western blot, immunofluorescence, Nissl staining, and qPCR were assessed post‐SAH. In vitro, hemin was introduced into HT22 cells to develop a model of SAH. Results. Stimulation of Nurr1 with AQ improved the outcomes and attenuated brain edema. Nurr1 was mainly expressed in neuron, and administration of AQ alleviated neuron injury in vivo and enhanced the neuron viability and inhibited neuron apoptosis and necrosis in vitro. Besides, AQ reduced the amount of IL‐1β+Iba‐1+ cells and inhibited the mRNA level of proinflammatory cytokines (IL‐1β and TNF‐α) and the M1‐like phenotype markers (CD68 and CD86). AQ inhibited the expression of MMP9 in HT22 cells. Furthermore, AQ reduced the expression of nuclear NF‐κB and Nurr1 while increased cytoplasmic Nurr1 in vivo and in vitro. Conclusion. Pharmacological activation of Nurr1 with AQ alleviated the neuron injury and neuroinflammation. The mechanism of antineuroinflammation may be associated with the Nurr1/NF‐κB/MMP9 pathway in the neuron. The data supported that AQ might be a promising treatment strategy for SAH.

Severity of odontogenic phlegmons affects local mucosal immunity

Safe methods for examining phlegmon of the maxillofacial area (MFA) have been extensively sought for, and assessing cytokine profile in the patient mixed unstimulated saliva may be used to control its development. There were enrolled 102 patients with maxillofacial phlegmon aged 25 to 75 years as well as age-matched 30 healthy subjects. Concentration of IL-1β, TNFα, TNFβ, IL-17, IFNγ, IL-4, IL-10, IL-22, TGF-β1 и TGF-β2 in mixed unstimulated saliva was measured. We found that patients with MFA inflammation vs. control group had significantly increased level of almost all cytokines except IFNγ. Additionally, we analyzed changes in saliva cytokine profile of patients with maxillofacial phlegmon, subdivided into groups by age, gender, and spread of the disease. It allowed to find that amount of IL-1β, TNFα, IL-17, IL-10, IL-22, TGF-β1 directly correlated with spread of the process, whereas level of IL-4, IFNγ, TGF-β2, on the contrary, was decreased with aggravated disease severity.

Resveratrol ameliorates inflammatory and oxidative stress in type 2 diabetic Goto-Kakizaki rats

Type 2 diabetes is associated with inflammatory and oxidative stress. Resveratrol, a naturally occurring diphenolic compound, was shown to improve glycemic control and alleviate metabolic disturbances in Goto-Kakizaki (GK) rats, a non-obese model of type 2 diabetes. However, in GK rats effects of resveratrol addressing inflammatory and oxidative stress were not explored. The present study aimed to determine anti-inflammatory and anti-oxidative properties of resveratrol in these rats. GK and Sprague-Dawley (SD) rats were divided into 4 groups: GK control, GK treated with resveratrol, SD control and SD treated with resveratrol. Resveratrol (20 mg/kg b.w.) was given once a day for 10 weeks. It was shown that contents of inflammatory markers, interleukin 6 (IL-6), interleukin 1 β (IL-1β), tumor necrosis factor α (TNF-α) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), were increased in the skeletal muscle of diabetic rats, but these effects were prevented by resveratrol therapy. Similarly, amounts of IL-1β and TNF-α were elevated in livers of GK rats; however, this rise was alleviated in resveratrol-treated animals. Moreover, the contents of inflammation-related factors (IL-6, IL-1β, TNF-α and NF-κB) were augmented in adipose tissue of GK rats; nevertheless, in this tissue resveratrol was ineffective. Resveratrol reduced also lipid peroxidation in the skeletal muscle, reduced activities of glutathione peroxidase in blood serum and catalase in the livers of GK rats. Our new findings show that resveratrol therapy results in relieving inflammatory and oxidative stress in GK rats, which may be largely associated with the alleviation of metabolic disturbances in this model of diabetes. Nevertheless, it was demonstrated that the efficacy of resveratrol action is tissue-specific.

Staphylococcus epidermidis from prosthetic joint infections induces lower IL-1β release from human neutrophils than isolates from normal flora.

The aim of this study was to test the hypothesis that Staphylococcus epidermidis isolated from prosthetic joint infections (PJIs) differs from S.epidermidis isolated from normal flora in terms of its capacity to induce activation of caspase-1 and release of IL-1β in human neutrophils. The amount of active caspase-1 was determined over 6h by detecting Ac-YVAD-AMC fluorescence in human neutrophils incubated with S.epidermidis isolates from PJIs (ST2) or normal flora. The amount of IL-1β was detected by ELISA in neutrophil supernatants after 6h of incubation. Mean IL-1β release was lower after incubation with S.epidermidis from PJIs compared to isolates from normal flora, but no statistically significant difference was found in active caspase-1. Substantial inter-individual differences in both active caspase-1 and IL-1β were noted. These results suggest that evasion of innate immune response, measured as reduced capacity to induce release of IL-1β from human neutrophils, might be involved in the predominance of ST2 in S.epidermidis PJIs, but that other microbe-related factors are probably also important.

Abstract P415: ACE N-domain Regulates High-glucose Mediated Interleukin-1 beta Production by Renal Epithelial Cells Independently From Angiotensin II Generation

Research studies demonstrated that interleukin (IL)-1β contributes to the development of diabetic nephropathy and hypertension. However, the origin and regulation of IL-1β synthesis during diabetic kidney injury are still unknown. Here, we hypothesize that renal epithelial cells produce IL-1β in response to a high glucose stress and that angiotensin converting enzyme (ACE) plays a key role in this process. To study this, we isolated proximal tubular (PT) epithelial cells from wild-type (WT) and mice lacking either the ACE N-domain (NKO) or the C-domain (CKO) catalytic activity. These cells were exposed to normal (5 mM) or high (30 mM) glucose for 24 hours. IL-1β produced by PT cells were assessed by ELISA and RT-PCR. High glucose induced WT PT cells to release significant amounts of IL-1β (from 5±1 to 70±6 pg/ml, p&lt;0.001; n=6). When WT PT cells were exposed to a high glucose media in the presence of an ACE inhibitor (lisinopril, 10 mM), IL-1β levels were significantly reduced (from 70±6 to 38±6 pg/ml, p&lt;0.01). In contrast, AT1 receptor blockade by losartan did not change the amount of IL-1β produced by WT PT cells. To determine which ACE domain is associated with IL-1β production, NKO and CKO PT cells were exposed to high glucose. Strikingly, NKO PT cells released lower amounts of IL-1β when exposed to high glucose compared to WT (NKO: 15±7 vs. WT: 79±9 pg/ml, p&lt;0.01, n=4). No differences were observed between WT and CKO PT cells. Since the ACE N-domain degrades the anti-inflammatory tetrapeptide N-acetyl-Ser-Asp-Lys-Pro (AcSDKP), we tested whether the lower IL-1β production in NKO PT cells was due to an accumulation of AcSDKP. For this, we pre-treated NKO PT cells with a prolyl endopeptidase inhibitor (S17092, 50μM) to prevent the production of AcSDKP. Notably, this treatment increased the IL-1β response to high glucose in NKO PT cells (2.1±0.3-fold increase, p&lt;0.01, n=4). Our data indicate that: 1) PT cells can sense and respond to high glucose by secreting IL-1β and 2) the absence of the ACE N-domain blunts the production of IL-1β through a mechanism that involves AcSDKP accumulation. In conclusion, ACE might contribute to the inflammatory response that underlays diabetic nephropathy independently from angiotensin II generation.

Interleukin‐1β has trophic effects in microglia and its release is mediated by P2X7R pore

Numerous neuroinflammatory conditions such as multiple sclerosis show enhanced expression of the purinergic P2X7 receptor (P2X7R) in the neuroinflammatory foci, where increased microglial activation is a co-existing feature. Activated microglia are known to release a number of bio-active substances such as interleukin 1β (IL-1β). Previous studies have linked P2X7R stimulation to the processing and release of IL-1β, but whether P2X7R channel or P2X7R pore is the predominant entity driving that release is unknown. Also the effect of released IL-1β on microglial activation and proliferation is unclear. The aim of this study was to characterise IL-1β release dynamics from microglia. Primary hippocampal rat cultures with electron microscopy and immunohistochemistry was used to delineate the sub-cellular localisation of IL-1β. FM1-43 fluorescent dye and confocal microscopy were used to quantify vesicular exocytosis from microglia expressing the pore forming P2X7R versus a non-pore forming point mutant, P2X7RG345Y. Amount of IL-1β in culture was quantified with enzyme-linked immunosorbent assay (ELISA). IL-1β intracellular processing was blocked with inhibition of caspase 1 and its extracellular form was neutralised with an IL-1β neutralising antibody. Microglial activation and proliferation was quantified immunohistochemically and with aid of confocal microscopy. Immunohistochemistry revealed P2X7R and IL-1β co-localisation in lysosomes. Vesicular exocytosis was higher in microglia expressing the pore forming P2X7R compared to those expressing the non-pore forming mutant. IL-1β levels were raised in cultures expressing the pore forming P2X7R and this cytokine was found to mediate the trophic effects of P2X7R pore in microglia. Inhibition of IL-1β production and function resulted in a significant decrease in P2X7R-mediated microglial activation and proliferation. The results reveal IL-1β as a mediator of microglial activation and proliferation. Blockade of P2X7R pore could serve as a therapeutic target in alleviating the degree of inflammation seen in conditions such as multiple sclerosis.

NLRP3 inflammasome activation is associated with proliferative diabetic retinopathy.

Innate immunity and dysregulation of inflammatory processes play a role in vascular diseases like atherosclerosis or diabetes. Nucleotide-binding domain and Leucine-rich repeat Receptor containing a Pyrin domain 3 (NLRP3) inflammasomes are pro-inflammatory signalling complexes that were found in 2002. In addition to pathogens and other extracellular threats, they can be activated by various endogenous danger signals. The purpose of this study was to find out whether NLRP3 activation occurs in patients with sight-threatening forms of diabetic retinopathy (DR). Inflammasome components NLRP3 and caspase-1, inflammasome-related pro-inflammatory cytokines IL-1β and IL-18, vascular endothelial growth factor (VEGF), acute-phase cytokines TNF-α and IL-6, as well as adaptive immunity-related cytokine interferon gamma (IFN-γ) were measured from the vitreous samples of 15 non-proliferative diabetic retinopathy (non-PDR) and 23 proliferative diabetic retinopathy (PDR) patients using the enzyme-linked immunosorbent assay (ELISA) method. The adaptor protein apoptosis-associated speck-like protein containing a CARD (ASC) was determined using the Western blot technique. Inflammasome components were present in the vitreous of DR patients. Along with VEGF, the levels of caspase-1 and IL-18 were significantly increased, especially in PDR eyes. Interestingly, clearly higher levels of NLRP3 were found in the PDR eyes with tractional retinal detachment (TRD) than from PDR eyes with fully attached retina. There were no significant differences in the amounts of IL-1β, TNF-α, IL-6, and IFN-γ that were detectable in the vitreous of both non-PDR and PDR patients. Our results suggest that NLRP3 inflammasome activation can be associated especially with the pathogenesis of PDR. The lack of differences in TNF-α, IL-6, and IFN-γ also alludes that acute inflammation or T-cell-mediated responses do not dominate in PDR pathogenesis.

Oral Hygiene in Children with Epilepsy: Effect of Interleukin-1β and VEGFLevels in Gingival Crevicular Fluid

Aim: Biochemical parameters in gingival crevicular fluid (GCF) are affected by some systemic diseases and poor oral hygiene. The aim of this study was to evaluate the total amounts of Interleukin-1β (IL-1β) and vascular endothelial growth factor (VEGF) in GCF in children with epilepsy. Material and methods: 80 children with epilepsy in free seizure period (Test group) and 80 healthy children (Control group) were evaluated. Gingival index (GI), plaque index (PI), probing depth (PD), clinical attachment level (CAL) was measured. GCF was collected and its volume was measured. The total amounts of IL-1β and VEGF in GCF were analyzed in Biochemistry Laboratory. Results: The biochemical (IL-1β and VEGF) and clinic parameters (GI, PI, PD and GCF volume) were significantly higher in the children with epilepsy compared with healthy children (p<0.0001). When the epilepsy patients were divided into groups according to the drug used in this study, there were no significant differences in GCF levels of IL-1β and VEGF among the drug groups. Conclusion: According to these results it may be suggested that patients need an optimal oral care in epilepsy.