Nuclear NF-κB Research Articles

Protective effects of chlorogenic acid on isoflurane-induced cognitive impairment of aged mice.

Postoperative cognitive dysfunction (POCD) is characterized by impairment in cognitive functions in patients following anesthesia and surgery. Chlorogenic acid (CGA) is a plant‐derived compound possessing numerous bioactive properties. The aim of this study was to investigate the therapeutic potential of CGA in isoflurane (ISO)‐induced cognitive dysfunction of aged mice, and further identify the mechanisms involved in the protective effects of CGA. A total of 80 male C57BL/6 mice, 20‐month‐old, were randomly divided into control group, isoflurane group (ISO), and ISO + 30 mg/kg CGA group and ISO + 60 mg/kg CGA. CGA was given orally once daily for 7 days to the mice and they were exposed to ISO (1.5%; 4 h). The open‐field and Morris water maze tests were used to investigate the cognitive function of mice. Pretreatment with CGA significantly attenuated ISO‐induced cognitive impairment. The levels of IL‐1β, TNF‐α, IL‐6, nuclear p65 NF‐kB, cleaved caspase‐3, and Bax were significantly increased, while the levels of IkBα and Bcl‐2 were decreased in the hippocampus 24 h after the ISO anesthesia. All the mentioned effects induced by ISO were reversed by CGA pretreatment. Furthermore, ISO exposure induced marked downregulation of SOD, CAT, HO‐1, and NQO‐1 and elevation of MDA and nuclear translocation of Nrf2 in the hippocampus tissue. All these parameters were reversed by CGA treatment. Importantly, the higher dose of CGA (60 mg/kg) showed a greater neuroprotective effect. In conclusion, these findings suggest that CGA attenuates the ISO‐induced cognitive impairment via its anti‐inflammatory, anti‐oxidative, and anti‐apoptotic properties in aged mice.

Dexmedetomidine attenuates perioperative neurocognitive disorders by suppressing hippocampal neuroinflammation and HMGB1/RAGE/NF-κB signaling pathway.

Surgical trauma can induce an inflammatory response in the central nervous system. Neuroinflammation is a crucial pathological mechanism of perioperative neurocognitive disorders (PND). Dexmedetomidine (Dex) is an alpha (α)-2 adrenoceptor agonist that is widely used in the perioperative period. Previous studies have shown that Dex has neuroprotection in various nerve injury models, but its role in PND remains unclear. Our study aimed to observe the neuroprotective effect of Dex pretreatment on postoperative cognitive change and explore the effects of hippocampal neuroinflammation, microglial polarization and HMGB1/RAGE/NF-κB signaling pathway involved in Dex on PND in rats. Rats were pretreated with Dex alone or in combination with yohimbine (α-2 adrenoceptor antagonist) before surgery. Behavioral tests results showed that Dex ameliorated surgery-induced cognitive impairment in rats. Nissl, immunohistochemistry and TUNEL-NeuN staining results indicated that Dex reduced hippocampus damage and neuronal apoptosis caused by surgery. Dex preconditioning reduced the expression of the proinflammatory cytokines IL-1β, TNF-α and IL-6 in hippocampus. Immunohistochemical and immunofluorescence results showed that Dex preconditioning inhibited the activation of glial cells induced by surgery. Western blot analysis showed that Dex preconditioning downregulated the expression of M1 phenotype markers (CD86 and iNOS), HMGB1, RAGE and nuclear NF-κB and upregulated the expression of M2 phenotype markers (Arginase 1 and CD206) and cytoplasmic NF-κB. Yohimbine could inhibit the neuroprotective effect of Dex. These results indicated that Dex pretreatment could improve postoperative short-term cognitive impairment, and the neuroprotective mechanism may involve the suppression of hippocampal neuroinflammation, regulation of M1/M2 polarization, and inhibition of HMGB1/RAGE/NF-κB signal transduction.

Network pharmacology and experimental validation identify the potential mechanism of sophocarpine for COVID-19.

Introduction. Coronavirus disease 2019 (COVID-19) has caused a serious threat to public health worldwide, and there is currently no effective therapeutic strategy for treating COVID-19.Hypothesis/Gap Statement. We propose that sophocarpine (SOP) might have potential therapeutic effects on COVID-19 through inhibiting the cytokine storm and the nuclear factor NF-κB signalling pathway.Aim. The objective was to elucidate the potential mechanism of SOP against COVID-19 through a network pharmacology analysis and its experimental validation.Methodology. The BATMAN-TCM database was used to identify the therapeutic targets of SOP, while the GeneCards and DisGeNET databases were used to identify the targets related to COVID-19. A protein-protein interaction (PPI) network was constructed from the STRING and analysed using Cytoscape software. Gene ontology (GO), Kyoto Encyclopaedia of Genes and Genomes (KEGG) and disease ontology (DO) enrichment analyses of the co-targets were performed using Metascape. Autodock 4.2.6 and Pymol software were applied for molecular docking. Levels of the proinflammatory cytokines IL-6, TNFα and IL-1β were measured by ELISA, while mRNA expression levels of intercellular adhesion molecule 1 (ICAM-1), vascular endothelial growth factor A (VEGFA) and IFN gamma (IFNG) were detected by real-time quantitative reverse transcription PCR. The protein levels of the molecules involved in the NF-κB signalling pathway were validated by western blot analysis.Results. A total of 65 co-targets of SOP and COVID-19 were determined. GO and KEGG enrichment analyses suggested that SOP affected COVID-19 by regulating the IL-17 signalling pathway, TNF signalling pathway and other signalling pathways. The PPI network and molecular docking showed that p65, ICAM-1 and VEGFA were key targets of SOP against COVID-19 and the underlying mechanism was validated in A549 cells in vitro. SOP attenuated the LPS-induced production of TNF-α and IL-6 and downregulated the LPS-induced mRNA expression of ICAM-1, VEGFA and IFNG. Mechanistically, SOP pretreatment inhibited the phosphorylation of p65 and facilitated the activation of Nrf2.Conclusions. SOP has a potential therapeutic effect on COVID-19 through multiple pathways and targets, and inhibits the production of pro-inflammatory cytokines and molecules involved in the NF-κB signalling pathway.

Anti-Inflammatory Mechanism of An Alkaloid Rutaecarpine in LTA-Stimulated RAW 264.7 Cells: Pivotal Role on NF-κB and ERK/p38 Signaling Molecules.

Lipoteichoic acid (LTA) is a key cell wall component and virulence factor of Gram-positive bacteria. LTA contributes a major role in infection and it mediates inflammatory responses in the host. Rutaecarpine, an indolopyridoquinazolinone alkaloid isolated from Evodia rutaecarpa, has shown a variety of fascinating biological properties such as anti-thrombotic, anticancer, anti-obesity and thermoregulatory, vasorelaxing activity. It has also potent effects on the cardiovascular and endocrine systems. Herein, we investigated rutaecarpine’s (Rut) anti-inflammatory effects in LTA-stimulated RAW macrophage cells. The Western blot and spectrophotometric results revealed that Rut inhibited the production of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and interleukin (IL)-1β in the LTA-induced macrophage cells. Successively, our mechanistic studies publicized that Rut inhibited LTA-induced phosphorylation of mitogen-activated protein kinase (MAPK) including the extracellular signal-regulated kinase (ERK), and p38, but not c-Jun NH2-terminal kinase (JNK). In addition, the respective Western blot and confocal image analyses exhibited that Rut reserved nuclear transcription factor kappa-B (NF-κB) by hindering inhibitor of nuclear factor κB-α (IκBα) and NF-κB p65 phosphorylation and p65 nuclear translocation. These results indicate that Rut exhibits its anti-inflammatory effects mainly through attenuating NF-κB and ERK/p38 signaling pathways. Overall, this result suggests that Rut could be a potential therapeutic agent for the treatment of Gram-positive bacteria induced inflammatory diseases.

The Role of SOCS2 Cytokine Signaling Suppressor in the Regulation of ProInflammatory Activity of Whole Blood Cells after Lower Respiratory Tract Infection

The aim of the investigation was to study the relationship between the content of whole blood in mononuclear leukocytes in pneumonia and in apparently healthy individuals of cytokine signaling suppressor 2 (SOCS2) with the production of cytokines (TNFα, TGFb, IFNα, IFNβ, IFNγ, IL-1β, IL-2, IL-4, IL-5, IL-10, IL-12, IL-17A, RAIL-1, RANTES) and individual factors of the NF-kB and JAK / STAT signaling pathways (NF-kB2, p65, p50, STAT1, STAT3, STAT5B, STAT6). Materials and research methods . The research material was mononuclear cells isolated from venous blood samples, as well as blood plasma of practically healthy individuals and patients with pneumonia. In nuclear-cytoplasmic lysates of mononuclear blood cells, the concentration of the components of the nuclear transcription factor NF-κB, p65, p50, NF-κB2, factors STAT1, STAT3, STAT5B, STAT6, and protein SOCS2, was assessed by enzyme immunoassay. We also determined the concentration of TNFα, IL-1β, TGFb, IFNα, IFNβ, IFNγ, IL-1β, IL-2, IL-4, IL-5, IL-10, IL-17A, RAIL-1, RANTES. The results of this study indicate that the stage of pneumonia convalescence is accompanied by dysregulation of the production of the main proinflammatory cytokines, manifested by a decrease in the level of TNFα, TGFb, RANTES, IL-4, IL-17A, IFNβ, IFNγ and an increase in the production of IL-2 and IFNα. Against this background, a decrease in the phosphorylation of the STAT3 and STAT4 factors was noted, as well as a decrease in the content of p50 and p65 proteins in MNCs. These changes were associated with an increased content of the SOCS2 factor in MNCs. The analysis showed that an increase in the content of SOCS2 in MNCs from the minimum level determined by the concentration corresponding to the 1st quartile of the sample (1.3 ng / ml) to the maximum, determined by the 4th quartile of the sample (1.7 ng / ml) is associated with a decrease in production IL-1β, IL-4, IL-4, IL-5, IL-10, IL-17A, TGFb, RANTES and IFNβ against the background of an increase in the level of INFα, INFγ and IL-2. Changes in cytokine production were accompanied by an increase in STAT5B, STAT4, and NF-kB2 levels and a decrease in STAT3 phosphorylation. a decrease in the content in the cell of the components of the nuclear transcription factor NF-κB, in particular, p50, p65. Conclusion . The peculiarities of the relationship of SOCS2 with the studied factors suggests that its high level helps to limit the production of proinflammatory cytokines, in particular those produced by type 2 T-helpers and Th17, stimulates an increase in ICC sensitivity to IL-2 and stimulation of type 1 T-helpers. These effects are realized due to an increase in the phosphorylation of the STAT5 and STAT4 factors, a decrease in the STAT3 activity, and a change in the ratio of the components p50, p65 and NF-κB2 of the nuclear transcription factor NF-κB in the cell.

Lupeol triggers oxidative stress, ferroptosis, apoptosis and restrains inflammation in nasopharyngeal carcinoma via AMPK/NF-κB pathway

Objective Nasopharyngeal carcinoma is a malignant tumor with high incidence in Asia. This study investigated the anti-tumor capacities of lupeol in nasopharyngeal carcinoma. Methods CCK-8 assay was employed to select the suitable concentration and intervention time of lupeol in 5–8 F and CNE1 cells. The anti-cancer impacts of lupeol were evaluated by flow cytometry, ROS generation, western blotting, ELISA, iron assay, lipid peroxidation, mitochondrial membrane potential (MMP), TUNEL, and immunohistochemistry assays. Additionally, levels of AMPK/NF-κB pathway-related proteins were tested by western blotting. Results Cell viability was notably decreased after administration of lupeol ≧ 20 μM. 20 μM and 40 μM lupeol induced cell apoptosis, enhanced oxidative stress and restrained immune response in nasopharyngeal carcinoma cells to some extent, as evidenced by the elevation of apoptotic rate, Bax and cleaved caspase-3 expression, ROS production and malondialdehyde level, and reduction of levels of Bcl-2, MMP, superoxide dismutase, TNF-α, IL-6 and IL-1β. Also, lupeol promoted the iron secretion and lipid peroxidation, the effects of which were reversed by ferroptosis inhibitor (Fer-1). The inhibitory impacts of lupeol at the doses of 20 μM and 40 μM on glutathione and GPX4 levels were observed. Importantly, lupeol significantly elevated AMPKα phosphorylation, and reduced the levels of p-IκBα and nuclear NF-κB p65. Rescue assay stated that siAMPK could neutralize the above impacts of lupeol. Moreover, lupeol suppressed tumorigenesis of xenografts in nude mice. Conclusion Lupeol exerted the anti-cancer impacts by inducing oxidative stress, ferroptosis and apoptosis, and suppressing inflammation via the AMPK/NF-κB pathway in nasopharyngeal carcinoma.

The ROCK-ezrin signaling pathway mediates LPS-induced cytokine production in pulmonary alveolar epithelial cells

BackgroundEzrin/radixin/moesin (ERM) proteins are members of the protein 4.1 superfamily and function as linkers that connect the actin cytoskeleton to the plasma membrane of cells. ERM also play critical role in the Lipopolysaccharide (LPS)-induced inflammatory response. However, the signaling mechanisms involved in this process remain unclear. In this study, we aimed to investigate the potential role of the rho-associated coiled-coil containing protein kinase (ROCK) pathway in LPS-induced ezrin phosphorylation and cytokine production in pulmonary alveolar epithelial cells.MethodsCultured A549 and HPAEpiC cells were treated with LPS. The expression and localization of ezrin in A549 and HPAEpiC cells were then analyzed by western blotting and immunoflurescence. Activation of RhoA/ROCK was assessed by western blotting and RhoA activity assays. The interaction of ezrin with Syk and myeloid differentiation factor 88 (MyD88)/IL-1R-associated kinase 1 (IRAK-1) was investigated by co-immunoprecipitation. The activation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) was measured with electrophoretic mobility shift assays and by western blotting. ELISA and western blotting were performed to detect the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and high mobility group box 1 protein (HMGB1) release into the culture supernatant, and cellular HMGB1 levels.ResultsLPS induced ezrin phosphorylation in a concentration- and time-dependent manner. The blockade of RhoA/ROCK inhibited LPS-induced ezrin phosphorylation and its translocation from the cytoplasm to the cell membrane. Co-immunoprecipitation assays further revealed that ezrin associated with Syk constitutively, but only associated with MyD88/IRAK-1 upon LPS challenge. Moreover, LPS-induced p38 and nuclear NF-κB activation was found to be ezrin dependent. The suppression of ezrin by siRNA or the blockade of ROCK activation with Y-27632 reduced the production of TNF-α, IL-1β, and HMGB1 in response to LPS.ConclusionsOur findings reveal a novel regulatory mechanism involving ezrin in the LPS-induced production of pro-inflammatory cytokines, and highlight the importance of the RhoA/ROCK-ezrin/Syk-MyD88/IRAK1 axis. Data presented in this manuscript provide novel insights into the signaling pathways activated in pulmonary alveolar epithelial cells by LPS.3MUhuZjAzSev1aZk4ypCqUVideo

Environmental Effects Of Monosodium Glutamate On (NF-κB) Levels In The Male Reproductive System Of Rats

Monosodium glutamate (MSG) has been identified as a food additive that Adversely affects MSG use on the reproductive efficiency by several mechanisms, this study was carried out to determine its role in stimulating the immune response in the reproductive system by estimating levels of NF-κB in serum and of its expression in reproductive organs. Thirty sex males rats were used in this study each included 12 male rats. Male rats A and B are receiving 60 and 120 mg/kg for 28 days respectively male rats of the control group were left without treatment for the period of the experiment. The levels of Nuclear factor-kappa NF-κB in serum and tissue fluid of testes and epididymis were measured. histological changes in addition to detection NF-κB expression were studying in testes and epididymis. Results of the present study recorded the toxic effect of MSG as it caused an elevation in the levels of NF-κB in serum, testicular, and epididymis tissue fluid. histological alternation in the reproductive organs was observed to represent detachment and vacuolation of the seminal epithelium, degeneration of spermatogenesis edema in a lumen in testicular tissues. Epididymal sections appeared to sever necrosis, degeneration in the epithelial layer and stereocilia, and the lumen of the epididymis. Also there was an increasing density of NFκB protein immunoreactivity in both testis and epididymis.

Abstract 205: Macrophage Rac1-IL-1β Signaling Axis Promotes Atherosclerotic Calcification

Coronary artery disease caused by atherosclerosis is a major cause of morbidity and mortality worldwide. Calcification of atherosclerotic plaque is a marker of atherosclerotic burden and worsening cardiovascular outcomes. Inflammation plays a role in both atherosclerotic progression and calcification, but the mechanisms of calcification are not entirely well-defined. We sought to understand the role of macrophage Rac1 signaling in the natural progression of atherosclerotic calcification in the context of an experimental model of hyperlipidemia. We developed a tamoxifen-inducible, myeloid specific Rac1 -deletion model ( Csf1r mercremer Rac1 fl/fl ApoE -/- ) to evaluate the impact of macrophage-specific Rac1 on atherosclerotic calcification. After 20 weeks of ad libitum feeding on a cholesterol-supplemented high fat diet, Rac1 -deleted animals demonstrated significantly decreased atherosclerotic calcification compared to wild-type mice, irrespective of modest changes in lipid metabolism as demonstrated by plasma analysis. Mechanistically, Rac1 -deleted bone marrow derived macrophages demonstrated reduced expression of the potent inflammatory cytokine, IL-1β, in response to a cholesterol-based inflammasome activation. Rac1 -deleted macrophages also demonstrated decreased nuclear NF-κB activity and lost binding of NF-κB to its seed site in the IL-1β promoter as evaluated by chromatin immunoprecipitation. We further demonstrated that Rac1 and NF-κB form a complex with each other upon innate inflammatory signaling activation using a proximity ligation assay. We confirmed the important functional role of the Rac1 nuclear localization sequence in the expression of IL-1β, which together with the proximity ligation assay, suggests a role for Rac1 in chaperoning NF-κB to the IL-1β promoter. The interaction between macrophage Rac1 and NF-κB may have implications on numerous inflammatory disease processes, and uncovers a novel signaling contribution of Rac1 as a transcriptional co-factor for IL-1β in the progression of atherosclerosis. Targeted inhibition of Rac1 may have potential as a therapeutic treatment to mitigate progression of vascular calcification.

Down-regulation of NRIP1 alleviates pyroptosis in human lens epithelial cells exposed to hydrogen peroxide by inhibiting NF-κB activation

Purpose: To investigate the role of nuclear receptor-interacting protein 1 (NRIP1) in oxidative stressinduced apoptosis and pyroptosis in cataract disease.Methods: Human lens epithelial cells (HLE-B3 cells) were exposed to hydrogen peroxide (H2O2). NRIP1 expression in hydrogen peroxide (H2O2)-treated HLE-B3 cells was determined by western blotting and quantitative reverse transcription polymerase chain reaction (qRT-PCR). CCK8 and EdU staining were used to assess cell viability. Flow cytometry and western blotting were used to assess pyroptosis.Results: NRIP1 was significantly up-regulated in HLE-B3 cells post-H2O2 incubation (p < 0.01). Hydrogen peroxide incubation reduced cell viability and proliferation of HLE-B3 cells, while NRIP1 knockdown enhanced cell viability and proliferation. NRIP1 silencing attenuated the H2O2-induced increase in NLRP3, N-terminal domain of gasdermin D, caspase-1, interleukin (IL)-1β, and IL-18 in HLEB3 cells, but suppressed the pyroptosis of H2O2-treated HLE-B3 cells. Hydrogen peroxide incubation down-regulated protein expression of cytoplasmic NF-κB and up-regulated nuclear NF-κB, while the expression of cytoplasmic NF-κB was increased and nuclear NF-κB was decreased in HLE-B3 cells by HLE-B3 interference.Conclusion: NRIP1 down-regulation represses apoptosis and pyroptosis of H2O2-treated human lens epithelial cells by inhibiting NF-κB activation, thus, providing a potential strategy to treat cataract disease.

Small-molecule IKKβ activation modulator (IKAM) targets MAP3K1 and inhibits pancreatic tumor growth

Activation of inhibitor of nuclear factor NF-κB kinase subunit-β (IKKβ), characterized by phosphorylation of activation loop serine residues 177 and 181, has been implicated in the early onset of cancer. On the other hand, tissue-specific IKKβ knockout in Kras mutation-driven mouse models stalled the disease in the precancerous stage. In this study, we used cell line models, tumor growth studies, and patient samples to assess the role of IKKβ and its activation in cancer. We also conducted a hit-to-lead optimization study that led to the identification of 39-100 as a selective mitogen-activated protein kinase kinase kinase (MAP3K) 1 inhibitor. We show that IKKβ is not required for growth of Kras mutant pancreatic cancer (PC) cells but is critical for PC tumor growth in mice. We also observed elevated basal levels of activated IKKβ in PC cell lines, PC patient-derived tumors, and liver metastases, implicating it in disease onset and progression. Optimization of an ATP noncompetitive IKKβ inhibitor resulted in the identification of 39-100, an orally bioavailable inhibitor with improved potency and pharmacokinetic properties. The compound 39-100 did not inhibit IKKβ but inhibited the IKKβ kinase MAP3K1 with low-micromolar potency. MAP3K1-mediated IKKβ phosphorylation was inhibited by 39-100, thus we termed it IKKβ activation modulator (IKAM) 1. In PC models, IKAM-1 reduced activated IKKβ levels, inhibited tumor growth, and reduced metastasis. Our findings suggests that MAP3K1-mediated IKKβ activation contributes to KRAS mutation-associated PC growth and IKAM-1 is a viable pretherapeutic lead that targets this pathway.

Post-COVID pain syndrome: a review of international observations

The novel coronavirus infection is commonly referred to as COVID-19, sometimes by the name of the causative agent of a respiratory viral infection, as SARS-CoV-2. Frequently, the course of COVID-19 is divided into three main periods: acute COVID-19 (up to 4 weeks), post-acute COVID-19 (from 4 to 12 weeks), post-COVID (post-COVID; from 12 weeks to 6 months). If a more protracted course of COVID (over 6 months) is discussed, the term “long-COVID” is used. All observations demonstrated a high incidence of pain syndromes of various localization in the post-COVID period and long-COVID. According to survey data, 92.3% of patients with COVID-19 reported musculoskeletal problems at the time of admission. Pain syndrome is observed in 56.3% of cases 1 month after hospitalization. Three months after COVID-19, myalgia was observed in 40.55% of cases, joint pain in 39.18%, back pain in 31.62%, and lower back pain in 24.74%. After 6 months, joint pain continues to be observed in 18.59% of patients, myalgia – in 15.09%, back pain – in 14.39%, lower back pain – in 11.23%. In 50.8% of cases, patients reported new-onset pain, of which 38.5% had pain of moderate severity (≥3 points on the visual analog scale). Patients with new-onset pain during COVID had worse quality of life indicators and a negative correlation with the pain syndrome severity, which significantly hampered recovery. Data from a meta-analysis that included 47,910 patients with long-COVID and with a protracted course of COVID indicate that 19% of them had pain in the joints of various localization. The direct cytopathic effect of SARS-CoV-2 and the systemic immune inflammation that occurs in response to infection cause damage to the joint tissue. According to the Guidelines for the Treatment of Patients with the Consequences of COVID-19, it is recommended to use slow acting structure-modifying drugs – SYSADOA – in the pharmacological treatment regimen for patients with osteoarthritis, among which parenteral forms of pharmaceutically standardized drugs – chondroitin sulfate (CS) and glucosamine sulfate (GS) are preferred. GS and CS are inhibitors of the signaling cascade of the nuclear factor NF-κB, which is involved in the realization of biological effects of a pro-inflammatory cytokine (tumor necrosis factor α), the excessive activity of which is associated with the cytokine storm in COVID-19.

Consumption of Cashew (Anacardium occidentale L.) Nuts Counteracts Oxidative Stress and Tissue Inflammation in Mild Hyperhomocysteinemia in Rats.

Hyperhomocysteinemia (HHcy) is a methionine metabolism problem that causes a variety of inflammatory illnesses. Oxidative stress is among the processes thought to be involved in the pathophysiology of the damage produced by HHcy. HHcy is likely to involve the dysfunction of several organs, such as the kidney, liver, or gut, which are currently poorly understood. Nuts are regarded as an important part of a balanced diet since they include protein, good fatty acids, and critical nutrients. The aim of this work was to evaluate the anti-inflammatory and antioxidant effects of cashew nuts in HHcy induced by oral methionine administration for 30 days, and to examine the possible pathways involved. In HHcy rats, cashew nuts (100 mg/kg orally, daily) were able to counteract clinical biochemical changes, oxidative and nitrosative stress, reduced antioxidant enzyme levels, lipid peroxidation, proinflammatory cytokine release, histological tissue injuries, and apoptosis in the kidney, colon, and liver, possibly by the modulation of the antioxidant nuclear factor erythroid 2–related factor 2 NRF-2 and inflammatory nuclear factor NF-kB pathways. Thus, the results suggest that the consumption of cashew nuts may be beneficial for the treatment of inflammatory conditions associated with HHcy.

Dietary resveratrol improved production performance, egg quality, and intestinal health of laying hens under oxidative stress

Resveratrol (RV) is associated with protection against oxidative stress to improve health, however the effect of RV in layers under oxidative stress (OS) is limited. The objective of this experiment was to investigate the negative effect of OS and protective effects of RV against OS in laying hens. 40 Lohmann layers (25-wk-old; BW = 1.44±0.10 kg) were allocated to four treatments in a 2 × 2 factorial arrangement with either RV (0 or 600 mg/kg) or intraperitoneal injection of tert-butyl hydroperoxide (tBHP) (0 or 800 μmol/kg BW) for 31 days. The results shown that the hens challenged with tBHP presented lower egg-laying rate, feed intake, feed efficiency and higher defective egg rate (P(tBHP)<0.05). The RV were also observed to attenuated egg laying rate and feed intake reduction together with decreased broken egg rate under t-BHP challenge (P(Interaction)≤0.01). The tBHP challenged layer demonstrated lower intestinal morphology (villus height in duodenum and jejunum), lower antioxidant enzymes activities [total superoxidase (SOD), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC)], and glutathione (GSH) levels and higher malondialdehyde (MDA) level] (P(tBHP)<0.05). Dietary RV increased jejunal SOD, GSH-Px and T-AOC activities, and reduced MDA concentration (P(RV) ≤0.05). Layers under tBHP challenge up-regulated mRNA expression of pro-inflammatory cytokine [interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α)] and nuclear factor NF-κB (P(tBHP)<0.05) in jejunum. Dietary RV supplementation down-regulated mRNA gene expression of IL-1β, IL-6, TNF-α and NF-κB (P(RV) ≤0.05). Dietary RV up-regulated mRNA expression of jejunal barrier-related proteins (claudin-1, claudin-2, mucin-1, and occludin) and ovarian reproductive hormone receptor [steroidogenic acute regulatory protein (StAR), androgen receptor (AR), estrogen receptor 1 (ESR1), and activin a receptor type 1 (ACVR1)] (P(RV) ≤0.05). Overall, the results indicate that tBHP induced oxidative stress to result in reducing production performance, intestinal health and induced ovarian inflammation; whereas dietary RV was able to maintain intestinal health and mitigate the negative impact of tBHP challenge on production performance and ovarian function.

ETS variant transcription factor 6 enhances oxidized low-density lipoprotein-induced inflammatory response in atherosclerotic macrophages via activating NF-κB signaling.

Objectives: Macrophages play a critical role in atherosclerosis by contributing to plaque development, local inflammation, and thrombosis. Elucidation of the molecular cascades in atherosclerotic macrophages is important for preventing and treating atherosclerosis. This study aims to deepen the understanding of the mechanisms that regulate the function of aorta macrophage in atherosclerosis. Methods: In the current study, the expression and function of ETS variant transcription factor 6 (ETV6) in aorta macrophages in a mouse atherosclerosis model. Aorta macrophages were enriched by flow cytometry. ETV6 expression was analyzed by quantitative RT-PCR. The role of ETV6 in macrophage-mediated pro-inflammatory response was evaluated both in vitro and in vivo after ETV6 silencing. Results: A remarkable elevation of ETV6 in aorta macrophages of atherosclerotic mice was observed. In addition, in vitro analysis indicated that oxidized low-density lipoprotein (oxLDL) up-regulated ETV6 in macrophages via the NF-κB pathway. ETV6 silencing suppressed oxLDL-induced expression of IL-1β, IL-6, and TNF-α in macrophages in vitro. However, ETV6 silencing did not impact the uptake of either oxLDL or cholesterol by macrophages. Furthermore, ETV6 silencing suppressed oxLDL-induced activation of the NF-κB pathway in macrophages, as evidenced by less phosphorylation of IKKβ and NF-κB p65, more cytoplasmic IκBα, and lower nuclear NF-κB p65. Moreover, ETV6 silencing inhibited the production of IL-1β and TNF-α in aorta macrophages in vivo. Conclusion: ETV6 supports macrophage-mediated inflammation in atherosclerotic aortas. This is a novel mechanism regulating the pro-inflammatory activity of atherosclerotic macrophages.

Targeting the Non-Canonical NF-κB Pathway in Chronic Lymphocytic Leukemia and Multiple Myeloma.

Simple SummaryThis study was designed to investigate the potential for targeting the NF-κB-inducing kinase, NIK, in two common B-cell malignancies, chronic lymphocytic leukemia (CLL) and multiple myeloma (MM). Using a selective NIK inhibitor, CW15337, we were able to demonstrate that cell lines and primary tumor cells were sensitive to the effects of NIK inhibition, whilst normal lymphocytes were significantly more resistant to its cytotoxic effects. Sensitivity to CW15337 was associated with the nuclear expression of the NF-κB subunit, p52. Importantly, tumor samples from a subset of poor prognosis CLL patients, with mutations in a gene called BIRC3, showed elevated p52 expression and were particularly sensitive to NIK inhibition. Furthermore, the combination of CW15337 and ABT-199 (venetoclax) reversed the drug resistance observed when treating tumor cells with ABT-199 alone. Our study shows the potential for targeting NIK in both CLL and MM.In this study, we evaluated an NF-κB inducing kinase (NIK) inhibitor, CW15337, in primary chronic lymphocytic leukemia (CLL) cells, CLL and multiple myeloma (MM) cell lines and normal B- and T-lymphocytes. Basal NF-κB subunit activity was characterized using an enzyme linked immunosorbent assay (ELISA), and the effects of NIK inhibition were then assessed in terms of cytotoxicity and the expression of nuclear NF-κB subunits following monoculture and co-culture with CD40L-expressing fibroblasts, as a model of the lymphoid niche. CW15337 induced a dose-dependent increase in apoptosis, and nuclear expression of the non-canonical NF-κB subunit, p52, was correlated with sensitivity to CW15337 (p = 0.01; r2 = 0.39). Co-culture on CD40L-expressing cells induced both canonical and non-canonical subunit expression in nuclear extracts, which promoted in vitro resistance against fludarabine and ABT-199 (venetoclax) but not CW15337. Furthermore, the combination of CW15337 with fludarabine or ABT-199 showed cytotoxic synergy. Mechanistically, CW15337 caused the selective inhibition of non-canonical NF-κB subunits and the transcriptional repression of BCL2L1, BCL2A1 and MCL1 gene transcription. Taken together, these data suggest that the NIK inhibitor, CW15337, exerts its effects via suppression of the non-canonical NF-κB signaling pathway, which reverses BCL2 family-mediated resistance in the context of CD40L stimulation.

Thyroid-Stimulating Hormone Inhibits Insulin Receptor Substrate-1 Expression and Tyrosyl Phosphorylation in 3T3-L1 Adipocytes by Increasing NF-κB DNA-Binding Activity.

Background Abundant evidence indicates that thyroid-stimulating hormone (TSH) levels are associated with insulin resistance in adipocytes. However, the potential mechanism of the association remains uncertain. The objective of this study was to determine the potential role of TSH in the suppression of insulin receptor substrate-1 (IRS-1) expression and IRS-1 tyrosyl phosphorylation, which might contribute to insulin resistance. Methods Mouse 3T3-L1 preadipocytes were differentiated into adipocytes. After treatment with 0.01, 0.1, and 1.0 mIU/ml bovine TSH, the TNF-α concentration in the medium was determined by enzyme-linked immunosorbent assay (ELISA). Nuclear factor-kappa B (NF-κB) DNA-binding activity was quantified by electrophoretic mobility shift assay (EMSA). IRS-1 levels in adipocytes were quantified by Western blotting, and tyrosine phosphorylation was measured by immunoprecipitation. Results TSH induced TNF-α secretion in a dose-dependent manner. There was a significant positive correlation between NF-κB DNA-binding activity and TNF-α secretion. This effect and correlation were weakened by BAY 11-7082 (a nuclear NF-κB inhibitor) and H89 (an inhibitor of cyclic adenosine monophosphate- (cAMP-) dependent protein kinase A (PKA)). Treatment of cultured adipocytes with TSH inhibited insulin-stimulated IRS-1 tyrosyl phosphorylation but promoted TSH-dependent secretion of TNF-α and activation of NF-κB DNA-binding activity. The effects of TSH were significantly inhibited by BAY 11-7082 and H89 and were completely blocked by the TNF-α antagonist WP9QY. Conclusion TSH inhibited IRS-1 protein expression and tyrosyl phosphorylation in 3T3-L1 adipocytes by stimulating TNF-α production via promotion of NF-κB DNA-binding activity. TSH might play a pivotal role in the development of insulin resistance.

Anti-inflammatory effects of fish bone fermented using Monascus purpureus in LPS-induced RAW264.7 cells by regulating NF-κB pathway.

Fish bones are the by-products of aquatic and fishery processing, which are often discarded. However, it has been considered having health-promoting by containing many essential nutrients. This study investigates the anti-inflammatory effect of fish bone fermented by Monascus purpureus (FBF) and the NF-κB pathway regulation mechanism in lipopolysaccharides (LPS)-induced RAW 264.7 cells. FBF has inhibited the production of PGE2 (prostaglandin E2), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in LPS-induced RAW264.7 cells. The FBF has significantly inhibited mRNA expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Moreover, FBF has suppressed activation of NF-κB (nuclear factor kappa-B) by increasing IκB mRNA expression and reduced of p65, p50 mRNA expression, as well as nuclear NF-κB DNA binding activity in LPS-induced RAW 246.7 cells. These findings demonstrate that FBF has inhibited LPS-induced inflammation by subsiding the activation of NF-κB in RAW 246.7 cells, implying that FBF could be employed as a promising natural product.

A Non-Nuclear NF-κB Modulates Alcohol Sensitivity But Not Immunity.

NF-κB proteins are well known as transcription factors important in immune system activation. In this highly conserved role, they contribute to changes in behavior in response to infection and in response to a variety of other insults and experiences. In some mammalian neurons, NF-κBs can be found at the synapse and translocate to the nucleus to alter gene expression when activated by synaptic activity. Here, we demonstrate that, in Drosophila melanogaster, NF-κB action is important both inside and outside the nucleus and that the Dif gene has segregated nuclear and non-nuclear NF-κB action into different protein isoforms. The DifA isoform is a canonical nuclear-acting NF-κB protein that enters the nucleus and is important for combating infection. The DifB variant, but not the DifA variant, is found in the central nervous system (mushroom bodies and antennal lobes). DifB does not enter the nucleus and co-localizes with a synaptic protein. In males and females, a DifB mutant alters alcohol behavioral sensitivity without an obvious effect on combating infection, whereas a DifA mutant does not affect alcohol sensitivity but compromises the immune response. These data are evidence that the non-nuclear DifB variant contributes to alcohol behavioral sensitivity by a nongenomic mechanism that diverges from the NF-κB transcriptional effects used in the peripheral immune system. Enrichment of DifB in brain regions rich in synapses and biochemical enrichment of DifB in the synaptoneurosome fraction indicates that the protein may act locally at the synapse.SIGNIFICANCE STATEMENT NF-κBs are transcription factors used by innate immune signaling pathways to protect against infection. Alcohol abuse also activates these pathways, which contributes to the addictive process and the health consequences associated with alcohol abuse. In the mammalian nervous system, NF-κBs localize to synapses, but it is axiomatic that they effect change by acting in the nucleus. However, for the Drosophila Dif gene, immune and neural function segregate into different protein isoforms. Whereas the nuclear isoform (DifA) activates immune genes in response to infection, the CNS isoform acts nongenomically to modulate alcohol sensitivity. Immunohistochemical and biochemical assays localize DifB to synapse-rich regions. Direct synaptic action would provide a novel and rapid way for NF-κB signaling to modulate behavior.

Bcl-3: A Double-Edged Sword in Immune Cells and Inflammation.

The NF-κB transcription factor family controls the transcription of many genes and regulates a number of pivotal biological processes. Its activity is regulated by the IκB family of proteins. Bcl-3 is an atypical member of the IκB protein family that regulates the activity of nuclear factor NF-κB. It can promote or inhibit the expression of NF-κB target genes according to the received cell type and stimulation, impacting various cell functions, such as proliferation and differentiation, induction of apoptosis and immune response. Bcl-3 is also regarded as an environment-dependent cell response regulator that has dual roles in the development of B cells and the differentiation, survival and proliferation of Th cells. Moreover, it also showed a contradictory role in inflammation. At present, in addition to the work aimed at studying the molecular mechanism of Bcl-3, an increasing number of studies have focused on the effects of Bcl-3 on inflammation, immunity and malignant tumors in vivo. In this review, we focus on the latest progress of Bcl-3 in the regulation of the NF-κB pathway and its extensive physiological role in inflammation and immune cells, which may help to provide new ideas and targets for the early diagnosis or targeted treatment of various inflammatory diseases, immunodeficiency diseases and malignant tumors.