IL-1β Gene Expression Research Articles

Gene expression changes in the cerebellum are associated with persistent post-injury pain in adolescent rats exposed to early life stress

Chronic pain develops following injury in approximately 20% of adolescents, at twice the rate in females than males. Adverse childhood experiences also increase the risk for poor health outcomes, such as chronic pain. Emerging literature suggests the cerebellum to be involved in pain processing, however detailed explorations into how the cerebellum contributes to pain are lacking. Therefore, this study aimed to characterise chronic pain outcomes and cerebellar gene expression changes following early life stress and injury in both sexes. The adverse childhood experience of neglect was modelled using a maternal separation (MS) paradigm, which was combined with a subsequent injury (mild traumatic brain injury (mTBI) or plantar incision surgery) in adolescent male and female Sprague-Dawley rats. We measured behavioural nociceptive sensitivity, systemic modulators of pain such as calcitonin gene-related protein (CGRP) and Substance P, as well as gene expression of IL1β, GFAP, GR, MR, GABRA1, CNR1, MAOA, and DAT1 in the cerebellum to examine associations between pain and neuroinflammation, the stress response, inhibitory neurotransmission, and monoaminergic function. We found increases in mechanical nociceptive sensitivity following plantar incision surgery. Sex differences were observed in anxiety-like behaviour and neuroinflammation, whereas systemic pain modulators showed cumulative effects with the addition of stressors. Most interestingly however, the increases in nociceptive sensitivity were associated with the suppressed expression of cerebellar genes that regulate stress, inhibition, cannabinoid function, and dopaminergic function, alongside sex-dependent distinctions for genes involved in inflammation and injury. This study highlights a novel link between nociception and molecular function in the cerebellum. Further investigation into how the cerebellum contributes to pain in males and females will facilitate novel therapeutic insights and opportunities.

Effect of Quinolinic Acid on Behavior, Morphology, and Expression of Inflammatory/oxidative Status in Rats' Striatum: Is Coenzyme Q10 a Good Protector?

Quinolinic acid (QUIN) is a toxic compound with pro-oxidant, pro-inflammatory, and pro-apoptotic actions found at high levels in the central nervous system (CNS) in several pathological conditions. Due to the toxicity of QUIN, it is important to evaluate strategies to protect against the damage caused by this metabolite in the brain. In this context, coenzyme Q10 (CoQ10) is a provitamin present in the mitochondria with a protective role in cells through several mechanisms of action. Based on these, the present study was aimed at evaluating the possible neuroprotective role of CoQ10 against damage caused by QUIN in the striatum of young Wistar rats. Twenty-one-day-old rats underwent a 10-day pretreatment with CoQ10 or saline (control) intraperitoneal injections and on the 30th day of life received QUIN intrastriatal or saline (control) administration. The animals were submitted to behavior tests or euthanized, and the striatum was dissected to neurochemical studies. Results showed that CoQ10 was able to prevent behavioral changes (the open field, object recognition, and pole test tasks) and neurochemical parameters (alteration in the gene expression of IL-1β, IL-6, SOD, and GPx, as well as in the immunocontent of cytoplasmic Nrf2 and nuclear p-Nf-κβ) caused by QUIN. These findings demonstrate the promising therapeutic effects of CoQ10 against QUIN toxicity.

A Systemic and Local Comparison of Senescence in an Acute Anterior Cruciate Ligament Injury-A Pilot Case Series.

Senescence, a characteristic of cellular aging and inflammation, has been linked to the acceleration of osteoarthritis. The purpose of this study is to prospectively identify, measure, and compare senescent profiles in synovial fluid and peripheral blood in patients with an acute knee injury within 48 h. Seven subjects, aged 18-60 years, with an acute ACL tear with effusion were prospectively enrolled. Synovial fluid and peripheral blood samples were collected and analyzed by flow cytometry, using senescent markers C12FDG and CD87. The senescent versus pro-regenerative phenotype was probed at a gene and protein level using qRT-PCR and multiplex immunoassays. C12FDG and CD87 positive senescent cells were detected in the synovial fluid and peripheral blood of all patients. Pro-inflammatory IL-1β gene expression measured in synovial fluid was significantly higher (p = 0.0156) than systemic/blood expression. Senescent-associated factor MMP-3 and regenerative factor TIMP-2 were significantly higher in synovial fluid compared to blood serum. Senescent-associated factor MMP-9 and regenerative factor TGFβ-2 were significantly elevated in serum compared to synovial fluid. Correlation analysis revealed that C12FDG++/CD87++ senescent cells in synovial fluid positively correlated with age-related growth-regulated-oncogene (ρ = 1.00, p < 0.001), IFNγ (ρ = 1.00, p < 0.001), IL-8 (ρ = 0.90, p = 0.0374), and gene marker p16 (ρ = 0.83, p = 0.0416). There is an abundance of senescent cells locally and systemically after an acute ACL tear without a significant difference between those present in peripheral blood compared to synovial fluid. This preliminary data may have a role in identifying strategies to modify the acute environment within the synovial fluid, either at the time of acute ligament injury or reconstruction surgery.

Inflammasome Activation and Pro-inflammatory Cytokine Genes Expression Following Exposure of PPRV in Vero Cells

Background: Peste des petits ruminants (PPR), a highly contagious and fatal disease of small ruminants, caused by PPR virus (PPRV). Detection of microbes is mediated by pattern recognition receptors (PRRs). These PRRs induce the assembly of a multi-protein signaling platform called the Inflammasome. It detects the pathogens and induces the synthesis of pro-inflammatory cytokines interleukins (IL) i.e. IL-1β and IL-18. The aim of this study was to assess the activation of inflammasome related genes, following exposure to PPRV in Vero Cells and was to assess the effect of activation of inflammasome on downstream pro-inflammatory cytokines IL-1β and IL-18 gene expression. Methods: Vero cells were infected with PPRV. The cells were incubated for different periods. Total RNA was reverse transcribed and used for amplification by quantitative real time PCR (qRT-PCR) of NLRP3, ASC, Caspase-1 and cytokines IL-1β and IL-18 gene expression. Result: It was revealed that 1 MOI of PPRV was appropriate for NLRP3 inflammasome activation. NLRP3 and ASC showed an increased level of expression at 2-4 hrs post inoculation (hpi). While, the Caspase-1 showed biphasic expression on 2-4 and 24 hpi, further, in the synchrony to it, IL-1β and IL-18 expression were also found to increase at 2-4 and 24 hpi.

Activation of distinct inflammatory pathways in subgroups of LR-MDS

Aberrant innate immune signaling has been identified as a potential key driver of the complex pathophysiology of myelodysplastic neoplasms (MDS). This study of a large, clinically and genetically well-characterized cohort of treatment-naïve MDS patients confirms intrinsic activation of inflammatory pathways in general mediated by caspase-1, interleukin (IL)-1β and IL-18 in low-risk (LR)-MDS bone marrow and reveals a previously unrecognized heterogeneity of inflammation between genetically defined LR-MDS subgroups. Principal component analysis resolved two LR-MDS phenotypes with low (cluster 1) and high (cluster 2) levels of IL1B gene expression, respectively. Cluster 1 contained 14/17 SF3B1-mutated cases, while cluster 2 contained 8/8 del(5q) cases. Targeted gene expression analysis of sorted cell populations showed that the majority of the inflammasome-related genes, including IL1B, were primarily expressed in the monocyte compartment, consistent with a dominant role in determining the inflammatory bone marrow environment. However, the highest levels of IL18 expression were found in hematopoietic stem and progenitor cells (HSPCs). The colony forming activity of healthy donor HSPCs exposed to monocytes from LR-MDS was increased by the IL-1β-neutralizing antibody canakinumab. This work reveals distinct inflammatory profiles in LR-MDS that are of likely relevance to the personalization of emerging anti-inflammatory therapies.

Potent In Vitro and In Vivo Effects of Stachys lavandulifolia Methanolic Extract against Toxoplasma gondii Infection.

MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to evaluate the in vitro effect of the SLME on T. gondii tachyzoites. Totally, 72 male BALB/c mice (40 mice for in vivo evaluation of SLME and 32 mice for its toxicity effects on liver and kidney serum enzymes) were used for the present investigation. At first, 40 mice were orally pre-treated with the SLME at doses of 25, 50, and 75 mg/kg/day for two weeks. Mice were checked daily, and the rate of survival and the mean number of tachyzoites were recorded. Liver lipid peroxidation (LPO) and nitric oxide (NO) levels, the effects on kidney and liver function, as well as the expression level of the proinflammatory cytokines such as interleukin-1β (IL-1β) and interferon-γ (IFN-γ), were studied by the quantitative real-time PCR. Flow cytometry analysis was performed on the effects of SLME on the detection of apoptotic and necrotic cells in T. gondii tachyzoites. The SLME at the concentrations 75 and 150 µg/mL completely killed the tachyzoites after 2 hr of incubation. SLME at 25, 50, and 75 mg/kg/day increased the survival rate of infected mice by the sixth, seventh, and eighth days, respectively. SLME also significantly (p < 0.05) decreased the LPO and NO levels and upregulated the IL-1β and IFN-γ mRNA gene expression levels, whereas no considerable change was observed in the serum level of kidney and liver enzymes. Flow cytometry analysis revealed the prompted early and late apoptosis after exposure to T. gondii tachyzoites with various concentrations of SLME. We found the relevant in vitro anti-Toxoplasma effects of SLME against T. gondii. Moreover, the results confirmed the promising in vivo prophylactic effects of SLME. SLME provokes the innate immune system, induces apoptosis, modulates the proinflammatory cytokines, and inhibits hepatic injury in infected mice. With all these descriptions, further surveys are required to support these findings and elucidate this plant's possible mechanisms of action.

The Role of NLRP3 Inflammasome in Obesity and PCOS-A Systematic Review and Meta-Analysis.

Inflammasomes have recently been implicated in the pathogenesis of several chronic inflammatory disorders, such as diabetes and obesity. The aim of this meta-analysis was to investigate the possible role of the NLRP3 inflammasome in obesity and polycystic ovarian syndrome (PCOS). A comprehensive search of electronic databases was conducted to identify studies investigating NLRP3 its related components (Caspase 1, ASC and IL-1β) in adipose tissue and/or blood from obese individuals compared to non-obese controls. Another search was conducted for studies investigating NLRP3 in PCOS women and animal models. The ssearched databases included Medline, EMBASE, Cochrane Library, PubMed, Clinicaltrials.gov, the EU Clinical Trials Register and the WHO International Clinical Trials Register. The quality and risk of bias for the included articles were assessed using the modified Newcastle-Ottawa scale. Data were extracted and pooled using RevMan software for the calculation of the standardized mean difference (SMD) and 95% confidence interval (CI). Twelve eligible studies were included in the obesity systematic review and nine in the PCOS review. Of the obesity studies, nine (n = 270) were included in the meta-analysis, which showed a significantly higher adipose tissue NLRP3 gene expression in obese (n = 186) versus non-obese (n = 84) participants (SMD 1.07; 95% CI, 0.27, 1.87). Pooled analysis of adipose tissue IL-1β data from four studies showed significantly higher IL-1β gene expression levels in adipose tissue from 88 obese participants versus 39 non-obese controls (SMD 0.56; 95% CI, 0.13, 0.99). Meta-analysis of adipose tissue ASC data from four studies showed a significantly higher level in obese (n = 109) versus non-obese (n = 42) individuals (SMD 0.91, 95% CI, 0.30, 1.52). Of the nine PCOS articles, three were human (n = 185) and six were animal studies utilizing PCOS rat/mouse models. All studies apart from one article consistently showed upregulated NLRP3 and its components in PCOS women and animal models. In conclusion, obesity and PCOS seem to be associated with upregulated expression of NLRP3 inflammasome components. Further research is required to validate these findings and to elucidate the role of NLRP3 in obesity and PCOS.

Limosilactobacillus johnsoni and Limosilactobacillus mucosae and Their Extracellular Vesicles Alleviate Gut Inflammatory Injury by Mediating Macrophage Polarization in a Lipopolysaccharide-Challenged Piglet Model

BackgroundLimosilactobacillus johnsoni (L. j) and Limosilactobacillus mucosae (L. m) can alleviate the inflammatory response. ObjectivesThis study aimed to elucidate the underlying mechanisms by which L. j- and L. m-derived extracellular vesicles (EVs) mitigate lipopolysaccharide (LPS)-induced intestinal injury. MethodsPiglets were assigned to 4 groups: oral phosphate-buffered saline inoculation for 2 wk prior to intraperitoneal injection of physiological saline or LPS, and oral L. j/L. m inoculation for 2 wk prior to intraperitoneal injection of LPS. The intestinal integrity, macrophage markers, cytokine levels, and microbiota were determined. The cytokine levels and macrophage phenotype were detected after L. j/L. m and their EVs were coincubated with macrophages. The levels of cytokines, tight junction proteins, and apoptosis were measured after intestinal epithelial cells were cocultured with macrophages. ResultsLPS challenge decreased jejunal villus length; expression levels of zonula occludens-1 (ZO-1), occludin, arginase-1 (Arg1), and interleukin (IL)-10; and number of CD163+ cells and increased the expression levels of inducible nitric oxide synthase (iNOS), IL-1β, IL-6, and tumor necrosis factor (TNF)-α compared with that in the control. L. j and L. m pretreatment rescued the aforementioned indicators compared with LPS challenge. Pretreatment of L. j and L. m and their EVs reversed the levels of IL-1β, IL-6, TNF-α, and IL-10 and the gene expression of iNOS and Arg1 in the LPS group in macrophages. Pretreatment with L. j and L. m-derived EVs increased ZO-1 and occludin mRNA expression and reduced IL-1β, caspase-3, and bax gene expression in intestinal epithelial cells of the coculture system. Enzyme-treated EVs were less effective than native EVs. ConclusionsThis study suggests that EVs secreted by L. j and L. m control inflammation by modulating macrophage polarization, thereby improving intestinal barrier function.

Unraveling Mechanisms of Autoimmune Skin Blistering: Applying Single-Cell Transcriptomics to Pemphigus B Cells

Unraveling Mechanisms of Autoimmune Skin Blistering: Applying Single-Cell Transcriptomics to Pemphigus B Cells

#3566 ACSS2 AGGRAVATES RENAL TUBULAR INFLAMMATION BY INDUCING THE DYSREGULATION OF FATTY ACID METABOLISM AND MITOCHONDRIAL OXIDATIVE STRESS IN DIABETIC MICE

Abstract Background and Aims Treatment options for diabetic nephropathy (DN) are currently limited. Chronic renal inflammation has been widely recognized as a major promoter of DN. The tubular epithelial cells are a major source of IL-1β in diabetic db/db mice. Inhibition of Acetyl CoA Synthase 2 (ACSS2) has been shown to promote high-fat diet-induced liver injuries, but the underlying mechanism of action of ACSS2 in diabetic renal injuries has not been elucidated. Method Streptozotocin (STZ) was intraperitoneally injected to establish the diabetic mouse model. We investigated the effects of ACSS2 on diabetes-induced renal tubular injury in transgenic mice with global acss2 knockout (acss2−/−). PAS staining was carried out to evaluate pathological injury of kidneys in mice. The renal tissue immunostaining was conducted to detect IL-1β expression and macrophage distribution in kidney tissues of mice. HK-2 cell culture model was achieved by high glucose stimulation in vitro. Real-time PCR analysis was applied to evaluate gene expression of inflammatory factors IL-1β, TNF-α, and MCP-1. Western Blot and immunofluorescence staining were conducted to the activation of the NLRP3 pathway and mitochondrial reactive oxygen species (ROS). Results ACSS2 was upregulated in STZ-induced diabetic renal tubular cells, which is significantly co-expressed in IL-1β-positive renal tubular cells of STZ-induced diabetic mice. The mice with acss2−/- exhibited decreased renal tubular inflammation and inhibited NOD-like receptor protein 3 (NLRP3) inflammasome activation as evidenced by elevated protein levels of NLRP3 and cleaved caspase-1 when compared with diabetic wild-type mice. The selective inhibition of ACSS2 protected HK-2 cells in vitro when exposed to a high glucose environment by downregulating NLRP3 inflammasome and ameliorating mitochondrial oxidative stress. Mechanistically, we showed that mitochondrial dysfunction preceded inflammatory response in vitro. The elimination of mitochondrial ROS using mitochondria-targeted antioxidants (mito-TEMPO) reversed HG-mediated NLRP3 inflammation activation in HK-2 cells, which recapitalized the phenotype observed in acss2-/- mice. We also found that cytoplasm ACSS2, which was increased after HG stimulation, induced fatty acid synthase (FASN) mediated overproduction of free fatty acid. Downregulating FASN using FASN-small interfering RNA restored mitochondrial function, and suppressed NLRP3 inflammasome activation. Conclusion In conclusion, diabetes-induced ACSS2 upregulation led to subsequent FASN-mediated dysregulation of fatty acid metabolism and mitochondrial ROS overproduction, eventually causing NLRP3 inflammasome activation and renal inflammation. Therefore, our findings established the role of ACSS2 in diabetes-induced inflammation of renal tubular epithelium and unveiled the underlying mechanism of its action. ACSS2 appears as a promising target for treating diabetic renal tubular epithelial inflammation.

Effect of autologous conditioned serum (ACS) on histological characteristics and expression of soft tissue IL-1β gene after horizontal ridge augmentation surgery.

Horizontal ridge augmentation surgeries are common procedures in periodontics. Histological changes in soft tissues affect the success of surgery in many ways. Autologous conditioned serum (ACS) increases interleukin-1 receptor antagonist (IL-1Ra) and causes inflammation modulation. Therefore, the present study aimed to investigate the effect of ACS on histological changes and gene expression of soft tissues after horizontal ridge augmentation surgeries. This double-blind split-mouth clinical trial was performed on patients needing implants with horizontal ridge augmentation (n=21). The control and intervention groups were considered split-mouth in two areas of the patients' oral soft tissues. A collagen membrane impregnated with ACS was used on the test side, and only a collagen membrane was used on the control side. After four months, histological changes in soft tissues, such as the amount of connective tissue collagen, fibroblast and inflammatory cell counts, and expression of the IL-1β gene, were evaluated. The fibroblast counts in the ACS group were significantly higher than in the control group. In addition, ACS caused a significant increase in the amount of collagen in the soft tissues compared to the control group (P<0.01). However, the number of inflammatory cells was similar in the two groups (P>0.05). IL-1β gene expression was not significantly different between the case and control groups. Under the limitations of the present study and based on the results of histological examinations, ACS increased the number of fibroblasts and the amount of collagen in soft tissues without affecting inflammatory cells (P=0.562).

Incomplete Polymerization of Dual-Cured Resin Cement Due to Attenuated Light through Zirconia Induces Inflammatory Responses.

Zirconia restorations are becoming increasingly common. However, zirconia reduces the polymerization of dual-cured resin cement owing to light attenuation, resulting in residual resin monomers. This study investigated the effects of dual-cured resin cement, with incomplete polymerization owing to attenuated light through zirconia, on the inflammatory response in vitro. The dual-cured resin cement (SA Luting Multi, Kuraray) was light-irradiated through zirconia with three thickness diameters (1.0, 1.5, and 2.0 mm). The light transmittance and the degree of conversion (DC) of the resin cement significantly decreased with increasing zirconia thickness. The dual-cured resin cement in 1.5 mm and 2.0 mm zirconia and no-irradiation groups showed significantly higher amounts of hydroxyethylmethacrylate and triethyleneglycol dimethacrylate elution and upregulated gene expression of proinflammatory cytokines IL-1β and IL-6 from human gingival fibroblasts (hGFs) and TNFα from human monocytic cells, compared with that of the 0 mm group. Dual-cured resin cement with lower DC enhanced intracellular reactive oxygen species (ROS) levels and activated mitogen-activated protein (MAP) kinases in hGFs and monocytic cells. This study suggests that dual-cured resin cement with incomplete polymerization induces inflammatory responses in hGFs and monocytic cells by intracellular ROS generation and MAP kinase activation.

Effect of antioxidant and anti-inflammatory on bioactive components of carrot (Daucus carota L.) leaves from Jeju Island

The present study evaluated the potential of carrot (Daucus carota L.) leaf, a non-edible part of carrots, which are among the most consumed vegetables worldwide. The antioxidant activities of 70% ethanol extract (EEC) and hot water extract (HEC) of carrot leaves were compared. The results revealed that the total polyphenol content, total flavonoid content, and DPPH and ABTS radical scavenging activities were higher in EEC than in HEC. Both extracts protected the cells against H2O2-induced toxicity and markedly reduced the levels of reactive oxygen species in RAW 264.7 cells. Moreover, pretreatment of RAW 264.7 cells with EEC and HEC prior to H2O2 (500 μM) exposure increased superoxide dismutase and glutathione peroxidase activities in these cells. Notably, EEC and HEC increased intracellular catalase activity by 36.77 and 6.39 times, respectively. Compared to HEC, EEC remarkably inhibited the production of lipopolysaccharide-induced nitric oxide and reduced the gene expression of IL-6, IL-1β, iNOS, COX-2, and TNF-α. Comparative analysis of the composition of the extracts using HPLC–UV suggested notably higher contents of catechin, chlorogenic acid, caffeic acid, rutin, quercetin, and cynaroside in EEC than in HEC. Collectively, these results imply that carrot leaves are a potentially beneficial natural source of antioxidants and anti-inflammatory compounds in functional foods.

(R)-ketamine attenuates neurodevelopmental disease-related phenotypes in a mouse model of maternal immune activation.

Infections during pregnancy are associated with an increased risk of neuropsychiatric disorders with developmental etiologies, such as schizophrenia and autism spectrum disorders (ASD). Studies have shown that the animal model of maternal immune activation (MIA) reproduces a wide range of phenotypes relevant to the study of neurodevelopmental disorders. Emerging evidence shows that (R)-ketamine attenuates behavioral, cellular, and molecular changes observed in animal models of neuropsychiatric disorders. Here, we investigate whether (R)-ketamine administration during adolescence attenuates some of the phenotypes related to neurodevelopmental disorders in an animal model of MIA. For MIA, pregnant Swiss mice received intraperitoneally (i.p.) lipopolysaccharide (LPS; 100µg/kg/day) or saline on gestational days 15 and 16. The two MIA-based groups of male offspring received (R)-ketamine (20mg/kg/day; i.p.) or saline from postnatal day (PND) 36 to 50. At PND 62, the animals were examined for anxiety-like behavior and locomotor activity in the open-field test (OFT), as well as in the social interaction test (SIT). At PND 63, the prefrontal cortex (PFC) was collected for analysis of oxidative balance and gene expression of the cytokines IL-1β, IL-6, and TGF-β1. We show that (R)-ketamine abolishes anxiety-related behavior and social interaction deficits induced by MIA. Additionally, (R)-ketamine attenuated the increase in lipid peroxidation and the cytokines in the PFC of the offspring exposed to MIA. The present work suggests that (R)-ketamine administration may have a long-lasting attenuation in deficits in emotional behavior induced by MIA, and that these effects may be attributed to its antioxidant and anti-inflammatory activity in the PFC.

An immunoregulator nanomedicine approach for the treatment of tuberculosis.

Introduction: A nanoparticle composed of a poly (lactic-co-glycolic acid) (PLGA) core and a chitosan (CS) shell with surface-adsorbed 1,3 β-glucan (β-glucan) was synthesized. The exposure response of CS-PLGA nanoparticles (0.1mg/mL) with surface-bound β-glucan at 0, 5, 10, 15, 20, or 25ng or free β-glucan at 5, 10, 15, 20, or 25ng/mL in macrophage in vitro and in vivo was investigated. Results: In vitro studies demonstrate that gene expression for IL-1β, IL-6, and TNFα increased at 10 and 15ng surface-bound β-glucan on CS-PLGA nanoparticles (0.1mg/mL) and at 20 and 25ng/mL of free β-glucan both at 24h and 48h. Secretion of TNFα protein and ROS production increased at 5, 10, 15, and 20ng surface-bound β-glucan on CS-PLGA nanoparticles and at 20 and 25ng/mL of free β-glucan at 24h. Laminarin, a Dectin-1 antagonist, prevented the increase in cytokine gene expression induced by CS-PLGA nanoparticles with surface-bound β-glucan at 10 and 15ng, indicating a Dectin-1 receptor mechanism. Efficacy studies showed a significant reduction in intracellular accumulation of mycobacterium tuberculosis (Mtb) in monocyte-derived macrophages (MDM) incubated with on CS-PLGA (0.1mg/ml) nanoparticles with 5, 10, and 15ng surface-bound β-glucan or with 10 and 15ng/mL of free β-glucan. β-glucan-CS-PLGA nanoparticles inhibited intracellular Mtb growth more than free β-glucan alone supporting the role of β-glucan-CS-PLGA nanoparticles as stronger adjuvants than free β-glucan. In vivo studies demonstrate that oropharyngeal aspiration (OPA) of CS-PLGA nanoparticles with nanogram concentrations of surface-bound β-glucan or free β-glucan increased TNFα gene expression in alveolar macrophages and TNFα protein secretion in bronchoalveolar lavage supernatants. Discussion: Data also demonstrate no damage to the alveolar epithelium or changes in the murine sepsis score following exposure to β-glucan-CS-PLGA nanoparticles only, indicating safety and feasibility of this nanoparticle adjuvant platform to mice by OPA.

Tisochrysis lutea F&amp;M-M36 Mitigates Risk Factors of Metabolic Syndrome and Promotes Visceral Fat Browning through β3-Adrenergic Receptor/UCP1 Signaling

Pre-metabolic syndrome (pre-MetS) may represent the best transition phase to start treatments aimed at reducing cardiometabolic risk factors of MetS. In this study, we investigated the effects of the marine microalga Tisochrysis lutea F&M-M36 (T. lutea) on cardiometabolic components of pre-MetS and its underlying mechanisms. Rats were fed a standard (5% fat) or a high-fat diet (20% fat) supplemented or not with 5% of T. lutea or fenofibrate (100 mg/Kg) for 3 months. Like fenofibrate, T. lutea decreased blood triglycerides (p < 0.01) and glucose levels (p < 0.01), increased fecal lipid excretion (p < 0.05) and adiponectin (p < 0.001) without affecting weight gain. Unlike fenofibrate, T. lutea did not increase liver weight and steatosis, reduced renal fat (p < 0.05), diastolic (p < 0.05) and mean arterial pressure (p < 0.05). In visceral adipose tissue (VAT), T. lutea, but not fenofibrate, increased the β3-adrenergic receptor (β3ADR) (p < 0.05) and Uncoupling protein 1 (UCP-1) (p < 0.001) while both induced glucagon-like peptide-1 receptor (GLP1R) protein expression (p < 0.001) and decreased interleukin (IL)-6 and IL-1β gene expression (p < 0.05). Pathway analysis on VAT whole-gene expression profiles showed that T. lutea up-regulated energy-metabolism-related genes and down-regulated inflammatory and autophagy pathways. The multitarget activity of T. lutea suggests that this microalga could be useful in mitigating risk factors of MetS.

The Alleviating Impacts of Quercetin on Inflammation and Oxidant-antioxidant Imbalance in Rats with Allergic Asthma.

Asthma is an inflammatory disease of the airways. We assessed the anti-inflammatory and antioxidative impacts of quercetin, a plant derivative, on inflammatory and oxidative indices in lung tissue and serum of rats with asthma.Asth ma was induced by ovalbumin. Rats were divided into 4 groups: control, asthma+vehicle (Receieved normal saline), asthma+dexamethasone, and asthma+quercetin. After asthma induction, quercetin (50 mg/kg) and dexamethasone (2.5 mg/kg) were injected intraperitoneally once daily for 1 week. On day 50, lung histopathology indices; inflammatory factors; tissue gene expression, including GATA Binding Protein 3 (Gata-3), Tbx21 (T-bet), Transforming growth factor-β (TGF-β), Il10 (IL-10), Il1b (IL-1β), Il6 (IL-6), Acta2 (α-SMA), and Tnf (TNF-α); and oxidative stress indices (malondialdehyde [MDA], catalase [CAT], glutathioneperoxidase [GPX], superoxide dismutase [SOD], and total antioxidant capacity [TAC]) in tissue and serum, were evaluated. The results showed that quercetin reduced Gata3, Tnf, Tgfb1, Il1b, and Acta2 gene expression and increased Tbx21 gene expression following asthma. Quercetin also improved oxidative stress by decreasing MDA levels and increasing TAC, CAT, SOD, and GPX levels in serum and lung tissue. Furthermore, quercetin decreased IL6 and TNFα levels and increased IL10 levels in lung tissue after asthma was treated with quercetin. Quercetin ameliorates oxidative stress and inflammation caused by asthma, especially at the tissue level. Therefore, quercetin can be considered a potent antiasthmatic agent.

Dietary Alpha-Monolaurin for Nile Tilapia (Oreochromis niloticus): Stimulatory Effects on Growth, Immunohematological Indices, and Immune-Related Gene Expressions

Alpha-monolaurin is reported to exhibit strong antiviral and antibacterial effects. This paves the way for its use as a new generation of feed additives. The experiment was carried out to examine the effects of the inclusion of alpha-monolaurin as a feed additive for Nile tilapia (Oreochromis niloticus) growth and immune response. Four diets were formulated to include alpha-monolaurin at 0, 2, 4, and 6 g alpha-monolaurin kg−1 diets and fed to the Nile tilapia (initial weight, 3.19 ± 0.11 g) for 70 days. Compared to those of the control group, the final body weight, weight gain rate, specific growth rate, and efficiency of feed of fish fed 4 and 6 g alpha-monolaurin kg−1 diets were (P ≤ 0.05) higher. The diet supplemented with a 2 g alpha-monolaurin kg−1 diet (P ≤ 0.05) improved endogenous amylase and lipase more than other treatments. The intestinal villus length, width, and goblet cell number were increased (P ≤ 0.05) in fish fed a 6 g alpha-monolaurin kg−1 diet. The same treatment displayed decreased (P ≤ 0.05) AST and ALT and increased total protein, albumin, and globulin. The highest levels of IgG and IgM were also noted in fish fed a 6 g alpha-monolaurin kg−1 diet. The highest values of hematocrit, hemoglobin, and red blood cell and white blood cell counts were seen in fish fed either a 4 or 6 g alpha-monolaurin kg−1 diet. The highest levels of thyroxine, triiodothyronine, and growth hormone were recorded in fish fed a 6 g alpha-monolaurin kg−1 diet. Likewise, the same treatment recorded the highest levels of SOD, CAT, GSH, and GPx but the lowest MDA value. Diets supplemented with 4 g or 6 g alpha-monolaurin kg−1 displayed the highest gene expressions of IFN-γ and IL-1β; however, HP70 genes were downregulated. In summary, the study showed that monolaurin may exert immunostimulatory effects on the immune system of the Nile tilapia by modulation of the host immune response and through metabolite production.

Elevated expression of the AIM2 gene in response to Helicobacter pylori along with the decrease of NLRC4 inflammasome is associated with peptic ulcer development.

Helicobacter Pylori (H. Pylori) cause peptic ulcer disease (PUD), but the inflammasome's role in PUD is not well understood. Therefore this study has investigated inflammasome compartment expression and IL-1β production in gastritis (G) and peptic ulcer disease. This study was based on gene expression of inflammasome compartments on stomach biopsies of 50 patients with PUD as cases and 50 individuals with gastritis as controls. The expression of NLRC4, ASC, IL-18, and serum IL-1β decreased in the PUD group compared to the control group. AIM2 gene expression increased, and NLRP12 gene expression decreased in H. pylori-seropositive positive (HP+ ) individuals compared to H. pylori-seronegative (HP- ) individuals. The G-HP+ subjects had higher serum IL-1β and AIM2 gene expression than G-HP- subjects but lower NLRP3 and NLRP12 gene expression. The PUD-HP+ had lower serum IL-1β, but higher AIM2 and IL-18 expression than PUD-HP- . The PUD-HP- patients had decreased IL-18 expression than G-HP- group. The PUD-HP+ had lower serum IL-1β and NLRC4 expression than G-HP+, while NLRP1 and NLRP3 were higher in expression in PUD-HP+ . The expression of caspase-1, NLRP3 and NAIP were correlated with IL-1β and IL-18. In conclusion, a decrease in NLRC4, IL-18, ASC genes, and IL-1β levels in PUD patients compared to gastritis may act in the development of PUD. H. pylori caused AIM2 induction and reduced NLRP12, indicating their contribution to bacterial responses. Decreased NLRC4 expression and IL-1β protein, together with enhanced NLRP1, and NLRP3 expression, promotes H. pylori to develop peptic ulcers.

Modeling the regulation of the Il12b locus by NF-κB dimers

Abstract The transcription factor NF-κB is composed of a family, which regulates the expression of various genes in immune and inflammatory responses. The NF-κB family consists of five members: RelA/p65, RelB, c-Rel, p50/p105, and p52/p100, which control the transcription of target genes by binding to regulatory κB sites in homo-or heterodimer forms. Unlike the widely known RelA, the mechanisms related to the regulation of target genes by c-Rel is less understood. Interleukin 12b(Il12b), which encodes the p40 subunit of IL12 and IL23, is one example of gene that depend on c-Rel for proper activation in macrophages. The differences in intrinsic binding properties of c-Rel- and RelA-containing dimers at various regulatory sites around Il12b can be a key to understand the mechanism of c-Rel-containing dimers’ role in Il12b regulation. Using c-Rel and Il12b reporter mice with fluorescence live-cell imaging, we observe the single-cell dynamics of nuclear c-Rel and Il12b gene expression in bone marrow-derived macrophages induced by NF-κB activating ligands. We analyze the live-cell imaging data and extract quantitative features of nuclear c-Rel and Il12b expression from the time-series data. For additional mechanistic insights of the Il12b gene regulation by NF-κB dimers, we propose a mathematical model of dynamic gene regulation which includes actions of RelA- and c-Rel-containing dimers at κB sites proximal and distal to the Il12b locus. Our combined experimental and mathematical modeling approach will reveal quantitative insights that may provide a more comprehensive understanding of regulatory mechanisms underlying Il12b gene expression employed by NF-κB dimers.