Expression Of NF-κB Research Articles

Berberine Inhibits the Disruption of the Blood-Brain Barrier and Glial Cell Activation in a Rat Model of Acute Hepatic Encephalopathy.

Hepatic encephalopathy (HE) is a complex neurological disorder in individuals with liver diseases, necessitating effective neuroprotective interventions to alleviate its adverse outcomes. Berberine (BBR), a natural compound with well-established anti-fibrotic and neuroprotective properties, has not been extensively studied in the context of glial activation under hyperammonaemic conditions. This study evaluates the neuroprotective potential of BBR in a thioacetamide (TAA)-induced HE rat model, focusing on its effects on glial activation and NLRP3 inflammasome signalling. Neurological impairments were assessed using open field tests and sensory analysis. Western blotting was performed to evaluate the expression of glial and neuronal markers, tight junction proteins and NLRP3 inflammasome components in the cortex and hippocampus. Histopathological and molecular changes were further examined using H&E, immunohistochemistry and immunofluorescence staining. BBR treatment significantly improved behavioural abnormalities and reduced systemic ammonia levels in TAA-exposed rats. It restored blood-brain barrier integrity, as evidenced by reduced tight junction protein degradation. BBR inhibited the expression of NLRP3 inflammasome markers, including caspase-1, IL-1β, ASC, and NF-κB, while reducing glial cell activation (IBA-1 and GFAP). Notably, BBR diminished NLRP3 expression in glial cells, indicating its potent anti-inflammatory effects. Additionally, BBR preserved neuronal integrity, as demonstrated by the maintained expression of MAP-2 and NeuN and reduced cleaved Gasdermin D levels. These findings suggest that BBR alleviates behavioural and molecular abnormalities in HE through NLRP3 inflammasome inhibition, highlighting its potential as a therapeutic agent for managing HE.

Effects of Seaweed Polysaccharide on the Growth and Physiological Health of Largemouth Bass, Micropterus salmoides

A seven-week trial was designed to evaluate the effects of dietary seaweed polysaccharide (SP) supplementation on the growth performance and physiological health of largemouth bass. The results reveal that the 0.05SP group showed the best growth performance. The mRNA expression levels of tor, 4ebp1, and igf1 genes were remarkably down-regulated in the 0.15SP and 0.2SP groups compared to the control group. The CAT activities were significantly increased in the 0.05SP and 0.1SP groups, and the GSH-Px activity was increased in the 0.15SP group. The expression of the immune response-related gene nfκb was significantly down-regulated in the 0.1SP group, and those of tnfα and il-8 were at the maximum in the control group. Moreover, the expression of il-10 in the 0.15SP and 0.2SP groups was significantly down-regulated. Furthermore, endoplasmic reticulum stress (ERS)-related expression of atf6 was the highest in the control group. Furthermore, the chopα and bax expression levels in the 0.15SP and 0.2SP groups were significantly down-regulated compared with other groups. In addition, the highest expression level of bcl-xl was observed in the 0.15SP group. Finally, the quadratic regression analysis of antioxidant, immune, and ERS core parameters (CAT, nf-κb, and bcl-xl) determined 0.06–0.11% to be the optimal SP supplemental level in largemouth bass diets.

Pink1/Parkin signaling mediates pineal mitochondrial autophagy dysfunction and its biological role in a comorbid rat model of depression and insomnia

Using a chronic unpredictable mild stress (CUMS) combined with multi-platform water environment sleep deprivation (SD) as an animal model, the occurrence and development of human depression combined with insomnia were simulated. The abnormal mitochondrial autophagy signaling caused by the putative kinase 1/Parkin E3 ubiquitin protein ligase (Pink1/Parkin) signaling pathway directly affects the normal secretion of melatonin by the pineal gland, which may explain the pathogenesis of depression combined with insomnia. This study aims to explore the depression-like behavior, sleep changes, central oxidative stress response, pineal mitochondrial autophagy damage, melatonin secretion, histopathological changes of the pineal gland, and the expression of Pink1/Parkin signaling-related factors in CUMS+SD rats. The results showed that the levels of reactive oxygen species (ROS) in cerebrospinal fluid of CUMS+SD rats significantly increased along with the inflammatory factors Interleukin-1β (IL-1β) and nuclear factor kappa-B (NF-κB) in cerebrospinal fluid. In addition, the number of pineal gland cells significantly decreased, cell boundaries became blurred, cell volume shrank, and apoptotic bodies appeared in the pineal gland tissue under HE staining, indicating pineal gland inflammation. Sleep deprivation further disrupted the levels of autophagy damage factors, including histamine (MDA), glutathione (GSH), and catalase (CAT), in the cerebrospinal fluid of CUMS + SD rats. Transmission electron microscopy of the pineal gland in CUMS+SD rats revealed damage to mitochondrial autophagy. The levels of 5-hydroxytryptamine (5-HT) and aromatic amine-N-acetyltransferase (AANAT) in the cerebrospinal fluid, as well as melatonin levels in the pineal gland, were significantly decreased. Additionally, the expression of IL-1β, NF-κB, Pink1, and Parkin in the pineal gland of CUMS+SD rats significantly increased. The expression of microtubule-associated protein 1 light chain 3-β (LC3), selective autophagy adaptor protein (P62), cytochrome c oxidase IV (COXIV), and mitochondrial outer membrane translocation enzyme 20 (TOM20) proteins downstream of the Pink1/Parkin signaling pathway was enhanced, while the expression of downstream brain-derived neurotrophic factor (BDNF), Beclin 1, and BCL2 interacting protein 3 (BNIP3) proteins was negatively regulated. Pink1/Parkin signaling may specifically respond to mitochondrial autophagy damage in the pineal gland, affecting the normal synthesis and secretion of melatonin in the pineal gland. In summary, mitochondrial autophagy damage in the pineal gland affects the normal secretion of melatonin in CUMS+SD rats, which is closely related to the specific autophagy signaling impairment of Pink1/Parkin pathway, which may mediate the occurrence of depression combined with insomnia.

Identification of Key Chondrocyte Apoptosis-Related Genes in Osteoarthritis Based on Weighted Gene Co-Expression Network Analysis and Experimental Verification.

Osteoarthritis (OA) is the primary cause of disability worldwide. Chondrocyte apoptosis has important implications for OA onset and progression. This work was designed to explore the mechanisms of chondrocyte apoptosis in OA and identify key chondrocyte apoptosis-related genes (CARGs). GSE32317 and GSE55235 datasets were acquired from the Gene Expression Omnibus (GEO) database. OA-associated module genes were determined via weighted gene co-expression network analysis (WGCNA) in GSE32317. CARGs were acquired from public databases. ClusterProfiler was employed for GO and KEGG analyses. Protein-protein interaction (PPI) network establishment was realized via the STRING database and Cytoscape, and the hub genes were screened by MCC, MNC, and DMNC algorithms of cyto-Hubba. The diagnostic values of the hub CARGs in OA in GSE55235 were verified via receiver operating characteristic (ROC) curve analysis. C28/I2 cells were stimulated with IL-1Β to establish the in vitro OA model. WGCNA identified 9,141 OA-related genes and 248 CARGs, resulting in 75 CARGs in OA. GO and KEGG analyses demonstrated that the 75 CARGs were primarily enriched in response to lipopolysaccharide, transcription regulator complex, and DNA-binding transcription factor binding, along with NF-kappa B and TNF signaling pathways. NFKB1 and ICAM1 were identified as the hub CARGs in OA through the three algorithms, which showed favorable prognostic values for OA. Notably, both bioinformatics analysis and in vitro assays revealed upregulated NFKB1 and ICAM1 expression in OA. NFKB1 and ICAM1 were the hub CARGs in OA, and might serve as diagnostic signatures and therapeutic targets for OA therapy.

Combined injection of pristane and Bacillus Calmette-Guerin Vaccine successfully establishes a lupus model with atherosclerosis.

Cardiovascular disease (CVD) induced by atherosclerosis (AS) is the main fatal complication of systemic lupus erythematosus (SLE). Establishing an appropriate animal model of SLE with AS is of great value for investigating the pathogenesis and therapeutic targets of SLE-CVD. In the present work, pristane was injected intraperitoneally into C57BL/6J mice to establish the SLE model and Bacillus Calmette-Guerin Vaccine (BCG) was injected intradermally one month later to enhance immunity and induce AS. Both pristane or pristane and BCG treated C57BL/6J mice exhibit a classical lupus-like phenotype which manifested as proteinuria and high levels of serum anti-ANA and anti-dsDNA autoantibodies, in conjunction with pathological changes in spleen and kidney, complement C3 deposition in the kidney, overactivation of T and B cells, a decreased proportion of splenic CD19+ B cells, and an increased frequency of CD19-CD138+ plasma cells, CD19+CD27+ memory B cells, CD3+CD4+ helper T (Th) cells, and CD3+CD4+CXCR5+PD-1+ T follicular helper cells. The combined pristane and BCG challenge aggravated AS in SLE mice, which had an enlarged thymus gland, an increased T cell proliferative vitality, an expanded Th cell pool, severe perivascular lymphocytes, and CD68+ macrophage infiltration, in addition to an intimal lipid deposition, and further elevation of Toll-like receptor 2 (TLR2), TLR4, and the transcription factor nuclear factor-κB (NF-κB) expression in kidney tissue and blood vessels when comparing with pristane or BCG injection alone. Pristane combined BCG challenge is able to establish a validated SLE-AS mouse model in C57BL/6J mice.

In vivo, in vitro, and in silico approaches in the detailed study of di-butyl phthalate (DBP), a plasticizer-induced lung fibrosis via Nrf-2/Keap-1/HO-1 pathway and its regulation.

The alveolar epithelium is a crucial barrier against external threats, yet it becomes a key player in initiating pulmonary fibrosis when compromised. Despite its importance, the intricate relationship between, DBP exposure and alveolar epithelial cell injury ensuing pro-fibrotic effects remains poorly understood. Phthalates, ubiquitous in nature, pose a significant risk to lung health upon inhalation, acting as immune triggers that cause airway inflammation and epithelial damage. We aimed to investigate the impact of intranasal administration ofDi-butyl Phthalate (DBP) inhalation, and its probable effects on normal and asthmatic lungs. DBP was administered via intranasal route in normal and OVA-induced asthmatic mice. DBP exposure enhanced oxidative stress and inflammatory parameters, leading to exacerbated asthmatic response and oxidative lung damage. Enhanced accumulation of immune cells, bronchial thickening, and collagen deposition was noted in histopathological investigations of DBP-exposed lung sections. Curcumin, a plant-derived molecule, significantly mitigated DBP-exposed asthma exacerbations by suppressing NF-κB expression and enhancing NRF2 levels via the Nrf-2/Keap-1/HO-1 signaling pathway. FACS analysis revealed increased CD11b+ cells (32%) in asthmatic mice which were reduced in the curcumin pre-treatment group (10.5%). Enhanced epithelial to mesenchymal transition (EMT) was noted in mice lungs and A549 cells where E-cadherin expression was reduced as compared to Vimentin, and α-SMA. Apart from aggravated airway inflammation, DBP exposure damages healthy lungs also. MMP-9/TIMP-1 ratios and collagen-1 levels were restored which were enhanced after DBP exposure. Moreover, antioxidant enzyme levels such as NQO-1, HO-1, and Catalase were significantly enhanced (p<0.01) and comparable to dexamethasone, a conventional corticosteroid. Notably, both dexamethasone and curcumin treatments effectively regulated the stimulation and accumulation of Nrf-2 in the nucleus, promoting antioxidant production and offering potential therapeutic benefits in mitigating pulmonary fibrosis. OVA and DBP alone caused DNA damage in the lung cells where increasedpercentage of damaged DNA movement in thetail, tail length, tail moment, and olive tail moment indicated severe damage in theDBP and OVA combined exposure strategies. Dexamethasone and Curcumin treatments reduced theextent of the DNA damage indicating anti-inflammatory and ant-oxidative potentials. Moreover, in silico studies are supportive of therapeutic potential of Curcumin and Dexamethasone in DBP-induced lung inflammation and fibrosis.

Impacts of Copper Deficiency on Oxidative Stress and Immune Function in Mouse Spleen.

Copper is an essential trace element crucial for enzyme synthesis and metabolism. Adequate copper levels are beneficial for maintaining the normal immune function of the spleen. Copper deficiency disrupts the metabolic processes within the spleen and impairs its immune function. This research examines the impact of copper deficiency on the spleen and the potential recovery following copper supplementation. Weaned mice underwent a 4-week copper-deficient diet, succeeded by 1-week of copper repletion via intraperitoneal copper sulfate injection. Histological examination was used to assess pathological changes in the spleen. Biochemical assays were performed to measure oxidative stress levels in the spleen. ELISA, qPCR, and Western blot were employed to examine alterations in inflammatory markers, immune indicators, and oxidative regulatory factors across various levels. Copper deficiency caused histological damage to the spleen, altered the expression of oxidative stress regulatory pathways (Nrf2, Keap1, and HO-1), and affected the expression of key inflammatory enzymes (iNOS, COX2) and transcription factor NF-κB, leading to oxidative damage. This was reflected by decreased levels of SOD, GSH, and T-AOC, along with increased levels of CAT and MDA. The levels of inflammatory cytokines IL-1β, TNF-α, IL-6, and IL-8 were notably increased. Copper supplementation significantly improved these changes. Copper deficiency leads to spleen tissue damage in mice, affecting the Nrf2 regulatory pathway and inducing oxidative damage. Subsequent copper supplementation with copper sulfate effectively ameliorates the damage caused by copper deficiency.

Down-regulation of interlinked inflammatory signalling cascades by ethanolic extract of Suaeda fruticosa Forssk. ex J.F. Gmel. attenuated in vivo inflammatory and nociceptive responses.

Juice and decoction of leaves of Suaeda fruticosa, a halophytic medicinal plant of Cholistan desert, is traditionally used to treat rheumatism. The current study was carried out to probe into in vivo anti-nociceptive, anti-inflammatory, and anti-arthritic potential of ethanolic extract of the whole plant of S. fruticosa (Et-SF)and its bioactive molecules. GC-MS screening of Et-SF revealed presence ofvarious bioactive compounds including phytol, thymol, n-hexadecanoic acid,farnesol, and 1-heptacosanol. DPPH in vitro radical scavengingassay demonstrated moderate antioxidant potential of Et-SF. Safety evaluation of Et-SF confirmed no lethal effects in female albino rats up to the singleoral dose of 5000mg/kg. In all invivo models, Et-SF was administered in three doses (125, 250, and 500mg/kg) and a single dose of flurbiprofen (FP) (10mg/kg). Et-SF significantly (p < 0.05) attenuated acute inflammation in carrageenan, histamine, and serotonin-induced rat paw oedema models in a time-dependent manner. Et-SF alleviated oedema, restored haematological parameters, and reduced severe pannus formation, inflammatory cell infiltration, and fibrous tissue proliferation inthe paws of CFA-induced arthritic rats.Moreover, treatment with thymol, farnesol and n-hexadecanoic acid alone and in combination also attenuated the arthritic progression in the arthritic rats indicating involvementof these compounds towards anti-arthritic potential of Et-SF. Et-SF and FP significantly (p < 0.05) down-regulated IL-1β, TNF-α, IL-6, NF-κB, and COX-2 mRNA expression, and up-regulated IL-4 and IL-10 mRNA expression in arthritic rats. Hot plate and acetic acid-induced writhing models results indicated the analgesic attributes of Et-SF in mice models. This study suggests that S. fruticosa ethanol extract may regulate the expression ofinflammatory markers involved in nociceptive, inflammatory, and arthritic disorders. Its phytochemicals could target multiple phases of these conditions at cellular and subcellular levels. Further research is needed to confirm this hypothesis.

Bioinformatics based exploration of the anti-NAFLD mechanism of Wang’s empirical formula via TLR4/NF-κB/COX2 pathway

BackgroundNonalcoholic fatty liver disease (NAFLD) has developed as a leading public wellness challenge as a result of changes in dietary patterns. Unfortunately, there is still a lack of effective pharmacotherapy methods for NAFLD. Wang’s empirical formula (WSF) has demonstrated considerable clinical efficacy in treating metabolic disorders for years. Nevertheless, the protective effect of WSF against NAFLD and its underlying mechanism remains poorly understood.MethodsThe NAFLD model was established using a 17-week high-sucrose and high-fat (HSHF) diet with 32 ICR mice. In assessing the therapeutic efficacy of WSF on NAFLD, we detected changes in body weight, viscera weight, biomarkers of glycolipid metabolism in serum and liver, transaminase levels and histopathology of liver with H&E and Oil Red O staining after oral administration. The chemical components in WSF were extensively identified and gathered utilizing the HPLC-Q-TOF/MS system, database mining from HMDB, MassBank, and TCMSP databases, alongside literature searches from CNKI, Wanfang and VIP databases. The forecast of network pharmacology approach was then utilized to investigate the probable mechanisms by which WSF improves NAFLD based on the performance of prospective target identification and pathway enrichment analysis. Besides, molecular docking was also conducted for the verification of combination activities between active components of WSF and core proteins related to NAFLD. In final, validation experiments of obtained pathways were conducted through ELISA, immunohistochemistry (IHC), and western blot (WB) analysis.ResultsPharmacodynamic outcomes indicated that WSF intervention effectively mitigated obesity, fat accumulation in organs, lipid metabolism disorders, abnormal transaminase levels and liver pathology injury in NAFLD mice (P < 0.05, 0.01). A total of 72 existent ingredients of WSF were acquired by HPLC-Q-TOF/MS and database, and 254 common targets (11.6% in total targets) of NAFLD and WSF were identified. Network pharmacology revealed that WSF presses NAFLD via modulating TNF, IL6, AKT1, IL1B, PTGS2 (COX2), and other targets, and the probable pathways were primarily inflammatory signaling pathways, as confirmed by molecular docking. Molecular biology experiments further conformed that WSF could decrease levels of inflammatory factors like IL-1β, IL-6 and TNF-α (P < 0.01) and expression of TLR4, NF-κB and COX-2 (P < 0.05, 0.01) in the liver.ConclusionWSF treatment effectively protects against lipid metabolism disorders and liver inflammation injury in HSHF diet-induced NAFLD mice, and its molecular mechanism might be via suppressing the TLR4/NF-κB/COX-2 inflammatory pathway to reduce the release of inflammatory cytokines in the liver.Graphical abstract

CD38 Coordinates with NF-κB to Promote Cochlear Inflammation in Noise-Induced Hearing Loss: the Protective Effect of Apigenin.

Noise exposure is one of the most common causes of sensorineural hearing loss. Although many studies considered inflammation to be a major contributor to noise-induced hearing loss, the process of cochlear inflammation is still unclear. Studies have found that activation of the NF-κB signaling pathway results in the accumulation of macrophages in the inner ear plays an important role in hair cell damage. In this study, tandem mass tag (TMT) technique was used to analyze the changes in basilar membrane proteome expression before and after acoustic injury. After noise exposure, the nicotinamide adenine dinucleotide (NAD) metabolism level was decreased, and the NF-κB signaling pathway was activated. The expression of CD38, the main NAD hydrolase in mammals, may directly lead to inflammation onset. Then, anakinra, an IL-1 receptor blocker, and apigenin, a CD38 inhibitor, were administered to animals to protect against noise-induced hearing loss. Our results showed that anakinra had little influence on the hearing threshold shift, while apigenin significantly reduce the threshold shift of hearing by inhibiting the expression of NF-κB and CD38 can be a promising target for protecting against noise-induced hearing loss.

Lactococcus lactis D4 Decreases NF-κB and α-SMA in Rat Models of Obstructive Jaundice

BACKGROUND: Obstructive jaundice, often due to choledocholithiasis or malignancies, leads to immune suppression, intestinal damage, and bacterial translocation, worsening outcomes. Some inflammatory mediators like nuclear factor kappa B (NF-κB), alpha-smooth muscle actin (α-SMA), and interleukin-6 (IL-6) are important in this process. Current treatments remain inadequate, highlighting the need for novel approaches. Probiotics, such as Lactococcus lactis D4 (LLD4), may help reduce inflammation and bacterial translocation, thus offering a potential therapeutic option. This study was conducted to evaluate the effects of LLD4 on NF-κB, α-SMA, and IL-6 in obstructive jaundice rat models.METHODS: This post-test randomized controlled study involved 15 male Wistar rats divided into three groups: sham, bile duct ligation (BDL), and BDL+LLD4 groups. The rats were maintained for 7–10 days, with the rats in BDL+LLD4 group received fermented milk containing LLD4 via gavage at a dose of 112 mg/20 gBW per day for 7 days. The expression levels of NF-κB, α-SMA, and IL-6 were analyzed using immunohistochemistry.RESULTS: Administration of LLD4 were able to significantly reduced NF-κB expression compared to the BDL group (40.20±21.276 vs. 53.60 ± 20.403) in obstructive jaundice rat models. Though not significant, BDL+LLD4 group showed lower α-SMA expression compared to BDL group (58.40±14.271 vs. 63.20±9.16). However, administration of LLD4 did not give any significant effect on IL-6 expression.CONCLUSION: LLD4 reduces inflammation in models of obstructive jaundice by lowering the NF-κB and α-SMA expression. This indicates that LLD4 might be potential as an adjunct therapy for reducing morbidity in obstructive jaundice cases.KEYWORDS: obstructive jaundice, bile duct ligation, Lactococcus lactis D4, NF-κB, α-SMA, IL-6

First Description of the Role of the Relationship Between Serum Amyloid P Components and Nuclear Factors/Pro-Cytokines During Critical Periods of Toxoplasmic Encephalitis.

Background/Objectives:Toxoplasma gondii (T. gondii), an obligate food-borne intracellular parasite, causes severe neuropathology by establishing a persistent infection in the host brain. We have previously shown that T. gondii infection induces severe neuropathology in the brain manifested by increased nitric oxide production, oxidative stress, glial activation/BBB damage, increased pro-inflammatory cytokine glia maturation factor-beta and induced apoptosis. Methods: The aim of this experimental study was to investigate the serum amyloid P (SAP) components, nuclear factor kappa B (NF-κB), interleukin-1 beta (IL-1β), caspase 1 (Casp 1), tumor necrosis factor-alpha (TNF-α) and complement 3 (C3) gene expressions on the 10th, 20th and 30th days after infection with T. gondii in the neuroimmunopathogenesis of toxoplasmic encephalitis (TE) in mouse brains by real-time quantitative polymerase chain reaction. The study also aimed to determine whether there was a correlation between the markers included in the study on these critical days, which had not previously been investigated. The mRNA expression levels of SAP components, NF-κB, IL-1β, Casp 1, TNF-α and C3 were examined. Results: The most notable outcome of this investigation was the observation that SAP components exhibited a 13.9-fold increase on day 10 post-infection, followed by a rapid decline in the subsequent periods. In addition, IL-1β expression increased 20-fold, while SAP components decreased 13-fold on day 20 after infection. Additionally, the TNF-α, Casp 1 and NF-κB expression levels were consistently elevated to above normal levels at each time point. Conclusions: This study identified SAP components, NF-κB, IL-1β, Casp 1 and TNF-α expressions as playing critical roles in TE neuroimmunopathogenesis. Furthermore, to the best of our knowledge, this is the first study to investigate SAP components during the transition from acute systemic infection to early/medium chronic and chronic infection and to explore the relationship between SAP components and other nuclear factors/pro-cytokines.

Exploring the effects of lactulose on lung-intestinal tissue-associated factors and the TLR4/NF-κB signaling pathway in COPD rats based on lung-gut axis theory.

Chronic obstructive pulmonary disease (COPD) is a systemic inflammatory disease impacting both the respiratory and gastrointestinal systems, with its pathogenesis closely linked to the lung-gut axis theory. In this study, we established a rat model of COPD using a fumigation method combined with intra-airway administration of lipopolysaccharide (LPS) to investigate the effects of lactulose on lung and intestinal tissues, focusing on related inflammatory markers and the TLR4/NF-κB signaling pathway. We further explored the therapeutic effects and mechanisms of lactulose on the lung-intestinal tissues in COPD rats, aiming to expand its potential application in chronic respiratory diseases. Subsequently, damage to lung and colon tissue was assessed using hematoxylin and eosin (HE) staining techniques, and ultrastructural changes in lung tissue cells were observed using transmission electron microscopy. Additionally, the study detected the expression of TNF-α, IL-1β, MUC5AC, TLR4, and NF-κB in lung tissue, along with the expression of MUC2, TLR4, and NF-κB in colon tissue, to evaluate the degree of tissue damage. The research findings indicated that the integrity of the airway and colon mucosal barrier in COPD rats was compromised. Intervention with lactulose significantly reduced pathological damage to lung and intestinal tissue and improved the microstructure of lung tissue. In addition, lactulose downregulated the expression of TNF-α, IL-1β, and MUC5AC in lung tissue, upregulated the expression of MUC2 in colon tissue, and inhibited the activity of the TLR4/NF-κB signaling pathway in lung and colon tissue, effectively mitigating inflammatory damage. Our findings demonstrated that lactulose effectively alleviates COPD-associated inflammation by inhibiting the TLR4/NF-κB signaling pathway within the lung-gut axis, underscoring its promise for treating multi-organ inflammation. This study provides a novel diagnostic and therapeutic perspective for the systemic management of COPD.

TACI Ig Fusion Protein Inhibits TLR4/MyD88/NF-κB Pathway Alleviates Renal Injury in IgA Nephropathy Rats.

To evaluate the impact of TACI fusion protein (TACI-Ig) on IgA nephropathy (IgAN) in rats, and to explore its mechanism and relationship with TLR4/MyD88/NF-κB pathway. Sprague Dawley(SD)rats were divided into six groups: control, model, TACI-Ig low dose (TACI-Ig-L), medium dose (TACI-Ig-M), high dose (TACI-Ig-H), and prednisone acetate (PAT) group. The control group and model group received physiological saline injections, while the TACI-Ig groups were administered doses of 7.18, 14.36, and 28.72 mg/kg of TACI-Ig, respectively. PAT group was pretreated with prednisone acetate. After 8 weeks, kidney weight/body weight ratios, 24-hour urine protein (24 h UP), serum creatinine (SCr), and blood urea nitrogen (BUN) levels were measured. Additionally, concentrations of B cell activating factor (BAFF), APRIL, and Gd-IgA1 were evaluated by using ELISA. Pathological changes in kidney tissues were scored, and TLR4, MyD88, NF-κB expression levels were detected through western blot (WB) and RT-qPCR. Results. Renal function assessments showed that the IgAN model group exhibited increased in 24 h UP, SCr, BUN, and elevated serum levels of BAFF, APRIL, Gd-IgA1, alongside higher TLR4/MyD88/NF-κB protein expression. TACI-Ig treatment significantly reduced proteinuria, SCr, BUN, levels of BAFF, APRIL, and Gd-IgA1 in IgAN rats. Pathologically, TACI-Ig ameliorated glomerular mesangial deposition and fibrosis. It also inhibited TLR4/MyD88/NF-κB protein expression, demonstrating anti-inflammatory and immune regulatory effects. TACI-Ig mitigates renal injury in IgAN rats by reducing inflammatory infiltration and IgA deposition and suppressing the pathway of TLR4/MyD88/NF-κB, offering data for developing effective treatments for IgAN.

The combination of microwave hyperthermia with TIPE2 impedes the growth of orthotopic colon cancer

Background Colon cancer (CC) is the main fatal disease of humans. Microwave hyperthermia (MH) is an adjuvant therapy for diverse cancers. Tumor necrosis factor-α induced protein-8-like 2 (TIPE2) is a tumor suppressor. However, the effect of MH combined with TIPE2 on CC remains unclear. Methods The orthotopic CC mouse model was constructed by mouse CC CT26-Luc cells, and mice were randomized into control, model (CT26-Luc), CT26-Luc + Vector, CT26-Luc + TIPE2, CT26-Luc + MH, and CT26-Luc + MH+TIPE2 groups (n = 6). Tumor growth pretreatment and post-treatment by in vivo fluorescence image analysis was detected. TIPE2 expression and cell transfection efficiency was detected by qRT-PCR and western blot. The pathological changes by HE staining, apoptosis by TUNEL staining, serum inflammatory factors by ELISA, TIPE2 expression by immunohistochemistry, and NF-κB signaling by western blot was performed. Results Paracancerous tissues showed higher TIPE2 expression than in CC tissues. CT26-Luc + TIPE2, CT26-Luc + MH, and CT26-Luc + MH+TIPE2 groups reduced tumor growth, tumor cell infiltration, and increased apoptosis. CT26-Luc + TIPE2 group had lower NF-κB, TNF-α, IL-1β, IL-6, p-p65, and p-IKK expression, and elevated TIPE2 and IkB expression, which was reversed by CT26-Luc + MH group. Moreover, CT26-Luc+MH+TIPE2 group showed opposite effects on the above factor expression of CT26-Luc+MH group. Conclusions Combination of MH with TIPE2 could impede CC tumor growth, providing scientific bases for its clinical application in CC treatment.

Piperazine ferulate impact on diabetes-induced testicular dysfunction: unveiling genetic insights, MAPK/ERK/JNK pathways, and TGF-β signaling.

Diabetic testicular dysfunction (DTD) poses a significant threat to male reproductive health. This study delves into the potential of piperazine ferulate (PF), a natural phenolic compound, in alleviating DTD and sheds light on its underlying mechanisms in rats. Animals were divided into the control, PF, diabetic, and diabetic plus PF groups. Diabetes was induced in rats with a single intraperitoneal (i.p.) injection of streptozotocin (STZ) at 50mg/kg. PF was administered at 50mg/kg/day via i.p. injection for four weeks. Significant changes in sexual behavior were observed in diabetic rats, which additionally revealed lower serum levels of testosterone, FSH, and LH. The abnormalities in sperm count, viability, motility, and morphology occurred along with the demonstrated suppression of genes and protein expression related to spermatogenesis. Atrophy of the seminiferous tubules and extensive degeneration and necrosis of the germ and Leydig cells were highlighted by histopathological examination. The testicular function of diabetic rats was significantly improved after PF administration, evidenced by normalized testicular histology, increased testosterone levels, and enhanced sperm quality. In addition to reducing inflammatory cytokines, COX2, and NF-κB expression, pf administration elevated the antioxidant levels and Nrf2/HO-1 expression. Furthermore, key signaling pathways involved in testicular degeneration are regulated by PF. It promoted cell survival and tissue repair by activating the protective TGF-β signaling pathway and attenuating the MAPK/ERK/JNK signaling cascade, which in turn reduced inflammation and apoptosis. PF suppressed the expression of INSL3, SPHK1, CD62E, ANGPTL2, and miR-148a-5p, while increasing the expression of testicular genes like HSD17B1, DAZL, and S1P, addressing DTD. This study highlights the potential of PF to restore testicular function and fertility in diabetic males by modulating genetic and signaling pathways.

Beta-Caryophyllene Augments Radiotherapy Efficacy in GBM by Modulating Cell Apoptosis and DNA Damage Repair via PPARγ and NF-κB Pathways.

Glioblastoma multiforme (GBM) is a highly aggressive brain malignancy with limited treatment options. Radiotherapy (RT) is often used for treating unresectable GBM; however, the outcomes are often limited due to the radioresistance of GBM. Therefore, the discovery of potential radiosensitizers to enhance GBM responses to RT is crucial. Beta-caryophyllene (BCP), a natural cannabinoid, promotes cancer apoptosis by upregulating the PPARγ signaling pathway and can cross the blood-brain barrier due to its lipophilic nature. This study aimed to evaluate the radiosensitizing potential of BCP in GBM cells. U87MG and GL261 cells and a GL261 tumor-bearing model were treated with RT, BCP, or both. Treatment efficacy was assessed using the MTT assay and tumor growth tracking, and the underlying mechanisms were investigated using western blotting, immunofluorescence staining, and other analyses. BCP synergistically enhanced the efficacy of RT in cell culture, as evidenced by the combination index determined through the MTT assay. This enhancement was mediated by the BCP-induced deceleration of DNA damage repair, as demonstrated by sustained γH2AX signal, upregulated PPARγ levels, and reduced expression of pAKT, pERK, and NF-κB, indicating apoptosis induction and inhibition of survival pathways. BCP significantly inhibited tumor growth in GL261 tumor-bearing mice with no discernible side effects. These findings indicate that BCP may serve as a potential radiosensitizer for improving RT outcomes in GBM by inhibiting DNA repair, inducing apoptosis, and suppressing anti-apoptotic and survival pathways.

Villin-1 regulates ferroptosis in colorectal cancer progression.

Colorectal cancer (CRC) is the second leading cause of cancer-related deaths worldwide. Despite extensive research, the mechanistic underpinnings driving CRC progression remain largely unknown. As a fundamental component of the brush border cytoskeleton, villin-1 (VIL1) acts as a marker for intestinal cell differentiation and maturation. Through a comprehensive transcriptomics analysis of eight studies (total sample: n = 1952), we consistently observed significant upregulation of VIL1 expression in CRC tumors compared with adjacent normal tissue. In our independent cohort, this notable upregulation has been further validated at both mRNA and protein levels in colon tumor tissues, relative not only to adjacent normal tissue but also to normal controls. Our data show that VIL1 promotes proliferation and migration while inhibiting apoptosis. Conversely, knockout of VIL1 suppresses proliferation and migration while inducing apoptosis. Mechanistically, we reveal that knocking out VIL1 activates ferroptosis and inhibits the migration of CRC cells, while overexpressing VIL1 yields the opposite effects, and vice versa. Additionally, VIL1 binds to Nuclear factor NF-kappa-B p105 subunit (NF-κB) and controls NF-κB expression. In vivo, overexpressing VIL1 inhibits ferroptosis, and induces the expression of NF-κB and lipocalin 2 (LCN2), thereby promoting CRC tumor growth. Thus, we have identified the VIL1/NF-κB axis as a pivotal regulator of CRC progression through ferroptosis modulation, unveiling VIL1 as a promising therapeutic target for CRC treatment via ferroptosis. Our study offers novel avenues for exploring the therapeutic potential of ferroptosis in CRC management, emphasizing the high potential of VIL1 in regulating colorectal tumorigenesis.

TRIM37 exacerbates cerebral ischemic injury by regulating the PPARγ/NF-κB pathway.

Ischemic stroke is the primary cause of mortality for individuals with disability worldwide. Tripartite motif 37 (TRIM37) plays multiple regulatory roles in various cellular processes. Our research aimed to investigate the effects of TRIM37 on the progression of ischemic stroke and its related mechanisms. Primary rat brain microvascular endothelial cells (BMECs) were treated with oxygen-glucose deprivation and reoxygenation (OGD/R) and then transduced with pShuttle-H1-TRIM37 shRNA plasmid, pShuttle-CMV-TRIM37 plasmid, or corresponding negative controls. The effects of TRIM37 were also explored in middle cerebral artery occlusion surgery-induced rat brain damage in vivo . Factor VIII staining showed the successful isolation of the primary BMECs. The OGD/R procedure significantly inhibited the cell viability and upregulated the TRIM37 expression in a time-dependent manner. In addition, OGD/R evidently increased the cell permeability, elevated the tumor necrosis factor alpha and intercellular adhesion molecule 1 levels, and upregulated the nuclear expression of nuclear factor (NF)-κB, but downregulated the peroxisome proliferator-activated receptors γ (PPARγ), zonula occludens-1, and cytoplasmic NF-κB expressions, which were reversed by TRIM37 knockdown. Furthermore, TRIM37 interacted with PPARγ and promoted its ubiquitination. The effects on cell permeability and inflammation induced by TRIM37 overexpression were reversed by PPARγ agonist. TRIM37 knockdown also inhibited middle cerebral artery occlusion-induced rat brain damage in vitro . TRIM37 may be a potential therapeutic target for ischemic stroke, and the knockdown of TRIM37 may improve brain injury by regulating the PPARγ/NF-κB pathway to modulate the inflammatory response.

P71 Bcl11b forms mutual connections with NF-κB1 and NF-κB2 in keratinocytes

Abstract Psoriasis is a chronic inflammatory skin condition characterized by complex molecular interactions. Our earlier data showed that Bcl11b expression is downregulated in psoriasis, and that proinflammatory cytokines act differently on Bcl11b associated activity of the NF-κB pathways. Given the critical role of NF-κB1 in the canonical pathway and NF-κB2 in the non-canonical pathway, it prompted us to elucidate the mutual regulatory connections between Bcl11b, NF-κB1, and NF-κB2, which are critical in the inflammatory response in psoriasis. To investigate the regulatory interplay between Bcl11b, NF-κB1, NF-κB2 and other key genes in the NF-κB pathways. We employed a series of knockout (KO) experiments using CRISPR–Cas9 technology to target Bcl11b, NF-κB1 and NF-κB2 in HaCaT keratinocytes. Gene and protein expression levels were assessed using qPCR and western blot, respectively. Specifically, we examined the impact of Bcl11b KO on NF-κB1 and NF-κB2 expression, followed by evaluating the reciprocal effects of NF-κB1 KO on NF-κB2 and BCL11B and the effect of NF-κB2 KO on NF-κB1 and Bcl11b. Additionally, we investigated the expression of related genes, including RelA, TNF-α, TAK1 RelB, NIK, TRAF2, TRAF3 and TRAF6. Bcl11b KO leads to a downregulation of both NF-κB1 and NF-κB2 at the RNA and protein levels, along with a downregulation of related pathway genes such as, RelA, TNF-α, RelB, and NIK. Conversely, both NF-kB1 KO and NF-kB2 KO conditions also demonstrated a mutual connection among Bcl11b and these two transcription factors. To gain a deeper understanding of the role of Bcl11b in the regulation of the NF-κB pathways, we evaluated the expression levels of several upstream regulators. Our study indicates an upregulation of TRAF2 and TRAF3 in Bcl11b KO conditions, corroborating the downregulation of the non-canonical pathway via NIK. Additionally, we observed a downregulation of TRAF6 under Bcl11b KO conditions, explaining the inactivation of the canonical NF-κB pathway. Interestingly, TAK1 has upregulated in Bcl11b KO cells, indicating a complex regulatory role of Bcl11b within the NF-κB signalling pathway. These data suggest a regulatory network where Bcl11b plays a crucial role in modulating both the canonical and non-canonical NF-κB pathways via TRAF2, TRAF3, and TRAF6. The mutual regulation between Bcl11b, NF-κB1 and NF-κB2 highlights potential therapeutic targets for modulating inflammatory responses in psoriasis.