Nuclear Factor Kappa Beta Research Articles

Ascorbic acid and calcitriol as alternative preventive strategies for myocardial damage in type 2 diabetes mellitus: an in vivo study using diabetic and atherosclerotic rat models

BACKGROUND: Type 2 diabetes mellitus can increase the production of reactive oxygen species (ROS) and genetic transcription factors, such as nuclear factor kappa beta (NF-κB). These phenomena may enhance the progression of atherosclerosis. Additionally, NF-κB can induce apoptosis of pancreatic beta cells and accelerate disease progression. OBJECTIVE: To investigate the role of calcitriol and ascorbic acid on NF-κβ expression in vivo in aortic and myocardial tissues of Wistar rats. METHODS: This randomized experimental study involved 24 male Wistar rats of the Rattus norvegicus strain, divided into four groups: NC (negative control), PC (positive control), AG (ascorbic acid group), and CG (calcitriol group). The study spanned 90 days, including a 30-day intervention period. Aortic and myocardial samples were processed into histological preparations and stained using immunohistochemical techniques. NF-κB expression was assessed using an intensity scoring method. RESULTS: The CG group demonstrated the lowest NF-κB immunoexpression in myocardial tissue (0.27 ± 0.08), followed by the AG (0.37 ± 0.05), NC (0.68 ± 0.08), and PC (1.13 ± 0.34) groups, with statistically significant 3sbetween the groups (p < 0.05). In aortic tissue, the CG group also exhibited the lowest NF-κB expression (0.30 ± 0.18), followed by the AG (0.50 ± 0.09), NC (0.97 ± 0.05), and PC (1.23 ± 0.38) groups, with statistically significant differences between the groups (p < 0.05). Similar trends were observed in the immunohistochemical staining of the NF-κB antigen in myocardial and aortic tissue samples. Calcitriol administration was more effective than ascorbic acid in reducing NF-κB expression in both myocardial (p < 0.05) and aortic tissues (p < 0.05). CONCLUSION: Both ascorbic acid and calcitriol reduce NF-κB expression in the aorta and myocardium, with calcitriol showing greater effectiveness than ascorbic acid. Keywords: Ascorbic acid, calcitriol, endothelial dysfunction, immunohistochemistry, antioxidant, NF-κB expression

Ameliorative role of camel protein hydrolysates diet against alkaline stress in Oreochrmis niloticus: Hematology, immune responses and their regulating genes expression, and histopathological assays.

This investigation looked at the ameliorative role of camel whey protein hydrolysates-diet (PH) in Oreochromis niloticus stocked under alkaline conditions. One hundred sixty fish (16.02 ± 0.14g) were allocated equally into four groups with four replications for 30 days. The first (control) and second (alkaline) groups were fed basal diets and maintained in fresh and alkaline water, respectively. The third and fourth groups were fed on a PH diet (basal diet containing 75g PH/kg) and maintained in fresh water and alkaline water, respectively. The hematology, immune-antioxidant indices, immune-regulatory genes, histopathological investigation of the spleen, and resistance to Aeromonas sobria were investigated. The results showed that the alkaline condition induced hematological disorders (lowered red blood cells, hemoglobin, packed cell volume, and white blood cell count) and immunosuppression (lowered phagocytic activity and index, lysozyme, nitric oxide, and complement 3) in the exposed fish. Alkaline exposure induced oxidative stress through elevation of the malondialdehyde and reduction in the antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase, glutathione S-reductase, and reduced glutathione). The immune modulatory genes (tolls like receptor-5, interleukin-1beta, interleukin-6, interleukin-8, interleukin-10, interleukin-17, nuclear factor kappa beta, and tumor necrosis factor-alpha) were down-regulated by exposure to alkaline conditions. The microscopic section of the spleen of the fish subjected to alkaline conditions showed notable hyperplasia of the melanomacrophage centers, besides vascular congestion, endothelial cell hypertrophy, and mild hypercellularity in the erythroid and lymphoid elements. In addition, few sections manifested more pronounced erythroid hyperplasia than the lymphoid one. The survival of the fish subjected to alkaline conditions was reduced during the A. sobria challenge. Feeding on a PH diet, the hematology was restored and the immune-antioxidant functions were modulated. Modulation of the immune-regulatory genes and increased survivability of the alkaline-exposed fish were noticed when fed on the PH diet. Consequently, we can recommend enriching the Nile tilapia diet with a 75g PH/kg diet especially when reared under alkaline conditions to support the immune functions of the fish.

Bioactive Compounds from Propolis on Bone Homeostasis: A Narrative Review.

This narrative review explores the potential effects of Propolis and its bioactive compounds on bone health. Propolis, a resinous product collected by bees, is renowned for its antimicrobial, anti-inflammatory, and antioxidant properties. Recent research emphasizes its positive role in osteogenesis, primarily through the modulation of osteoclast and osteoblast activity via molecular pathways. Key mechanisms include reducing inflammatory cytokines, protecting against oxidative stress, and upregulating growth factor essential for bone formation. While compounds such as Caffeic Acid Phenethyl Ester, Apigenin, Quercetin, and Ferulic Acid have been well-documented, emerging evidence points to the significant roles of less-studied compounds like Pinocembrin, Kaempferol, p-Coumaric acid, and Galangin. This review synthesizes the current literature, focusing on the mechanisms by which these bioactive compounds influence osteogenesis. Firstly, it explores the techniques for characterizing bioactive compounds presented in propolis, the chemogeographic variations in its composition, and the effects of both crude extracts and isolated compounds on bone tissue, offering a comprehensive analysis of recent findings across different experimental models. Further, it discusses the effects of Propolis compounds on bone health. In summary, these compounds modulate signaling pathways, including nuclear factor kappa beta, wingless-related integration site, mitogen-activated protein kinase, vascular endothelial growth factor, and reactive oxygen species. These pathways influence the receptor activator of nuclear factor kappa-β/receptor activator of nuclear factor kappa-β ligand/osteoprotegerin system, fostering bone cell differentiation. This regulation mitigates excessive osteoclast formation, stimulates osteoblast activity, and ultimately contributes to the restoration of bone homeostasis by maintaining a balanced bone remodeling process.

Vitamin D3 mitigates aspirin-induced gastric injury by modulating gastrokines, E-cadherin, and inhibiting NLRP3 and NF-κB/MMP-9 signaling pathway.

The prevalence of gastric ulcers has grown significantly in the modern era affecting 10 % of global population. Aspirin downregulates gastrokines 1(GKN1) expression in gastric mucosa and GKN1 down-regulation results in gastric cancer. Vitamin D3 (Vit.D3) has anti-inflammatory and antioxidant effects. Study the gastroprotective impact of Vit.D3 following aspirin-induced gastric injury in relation to gastrokines and investigate the possible underlying mechanisms. 24 rats were divided into 4 groups: control, Vit.D3 supplemented normal, aspirin-induced gastric injury, and Vit.D3 supplemented gastric injury groups. Some oxidative stress markers with gene expression of GKN1&2, mucin 5AC (Muc5ac) and NLR family pyrin domain containing 3 (NLRP3) in the gastric tissue were done. Histopathological and immunohistochemical study of E-Cadherin, nuclear factor kappa beta (NFκB), and metalloproteinase-9 (MMP-9) in the stomach mucosa were identified. Vit.D3 supplementation significantly upregulated E-Cadherin, GSH, GKN1and Muc5ac in the gastric tissue. Also, it improved the morphology, histology of gastric tissue, by alleviating oxidative stress and NFκB, MMP-9 and down regulation of inflammasome (NLRP3). Vitamin D3 has a potential protective effect against aspirin -induced gastric injury via upregulating gastrokine1 and E-cadherin and down regulation of NFKB/MMP-9 signaling pathway and NLRP3 inflammasome.

Red fruit (Pandanus conoideus Lam) oil ameliorates streptozotocin-induced diabetic peripheral neuropathy by targeting the oxidative and inflammatory pathways in the spinal cord in a rat model

Diabetic peripheral neuropathy (DPN) is a common comorbid in diabetic patients. Oxidative stress and inflammation are key to DPN’s etiology, making them possible therapy targets. Red fruit oil (RFO) may treat DPN due to its antioxidant and anti-inflammatory properties. The RFO effect in the streptozotocin (STZ)-induced DPN model was examined in terms of spinal cord oxidative and inflammatory functions. STZ 55 mg/kg BW intraperitoneally caused DPN. Seven weeks after induction, rats received vehicle, pregabalin, or RFO at 0.3, 0.6, or 1.2 mL/kg BW for three weeks. Post-treatment thermal hyperalgesia and cold allodynia were examined. Measurements of spinal levels of malondialdehyde (MDA), catalase, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), inducible nitric oxide (iNOS), and nuclear factor-kappa beta (NFκB) were used to measure oxidative and inflammatory indices. In addition, spinal histology was examined. All metrics improved after three weeks of RFO treatment. RFO at 0.6 and 1.2 mL/kg BW significantly reduced MDA, TNF-α, IL-6, iNOS, and NFκB while raising catalase levels. These matched thermal and cold stimulus latency improvements. Additionally, STZ-induced spinal cellular damage was reduced. This study suggests that RFO may be an alternate DPN treatment.

Melatonin ameliorates inflammation and improves outcomes of ischemia/reperfusion injury in patients undergoing coronary artery bypass grafting surgery: a randomized placebo-controlled study

To investigate the protective role of high dose melatonin concerning myocardial I/R injury and inflammation in patients undergoing on-pump coronary artery bypass grafting (CABG) surgery by evaluating IR/inflammatory biomarkers and clinical outcomes. This was a prospective; randomized; single-blinded placebo-controlled study conducted at cardio-thoracic surgery department of the Academy of the Cardiovascular and Thoracic Surgery, Ain Shams University. Eligible patients were randomly allocated to; melatonin-treated group (MTG) or placebo-treated group (PTG). The MTG (n = 17) received 60 mg/day melatonin capsules daily starting 5 days before surgery in addition to the standard of care. PTG (n = 17) received placebo also 5 days before surgery plus standard of care. The levels of nuclear factor kappa beta (NF-κb) (primary outcome), tumor necrosis factor (TNF-α), cardiac troponins I, and IL-6 levels were all assessed for both groups at five time points: baseline before melatonin or placebo administration (T0), before cross-clamp application(T1), 5 min after cross-clamp removal(T2), 6 h after cross-clamp removal(T3) and 24 h after cross-clamp removal(T4). Blood pressure was assessed at baseline, pre-operative and 24-hours post-operative. The Quality of recovery-40 score (QOR-40) was assessed for both groups on day 4 after surgery. TNF-α levels decreased in the MTG at T1(p = 0.034) versus PTG. At T2(p = 0.005), and T3(p = 0.04), TNF-α significantly increased in PTG versus MTG. Troponins significantly increased in PTG at T3 (p = 0.04) versus MTG. NF-κB levels declined at T1 (p = 0.013) and T2 (p = 0.0001) in MTG compared to PTG. IL-6 significantly increased in PTG versus MTG at T3 (p = 0.04). The QOR-40 score significantly decreased in MTG versus PTG. MTG had statistically significant decrease in DBP compared to the placebo group (p = 0.024). MTG had a statistically significant shorter intubation time than did the placebo group (p = 0.03). Melatonin 60 mg was well-tolerated without any reported side effects. Our findings suggested that melatonin could ameliorate myocardial I/R injury after on-pump CABG and that this outcome was essentially correlated to its antiapoptotic and anti-inflammatory effects. Trial registration: ClinicalTrials.gov registration number NCT05552586, 9/2022.

Effect of zinc or copper supplementation on the efficacy and sustainability of botulinum toxin A “Botox” injection in masseter muscle of albino rats

ObjectivesThis study aimed to evaluate whether oral zinc or copper supplementation affected the efficacy and sustainability of botulinum toxin-A (BTX-A) injection in masseter muscle of albino rats. Materials and methods32 adult male albino rats were allocated equally into four groups: group I (control), group II received 10U BTX-A injection, group III received 10U BTX-A injection + zinc (1 mg, 4 days pre-injection), and group IV received 10U BTX-A injection + copper (0.04 mg, 7 days post-injection). Rats were euthanized at 2 and 12 weeks (4 rats per subgroup) after injection. The masseter muscle was examined via histological, histochemical, histomorphometrical and real-time polymerase chain reaction (qRT-PCR) analyses. ResultsThe histopathological results of the BTX-A group showed atrophied muscle fibers with increased atrophy with time compared to the control group. The BTX-A + zinc group displayed more atrophy compared to BTX-A group. Conversely, the BTX-A + copper group demonstrated improved histology of muscle fibers compared to BTX-A and BTX-A + zinc groups. Histomorphometric analysis of Masson trichrome staining at 2 and 12 weeks revealed that collagen area percentage was the highest in the BTX-A + copper and control groups, followed by BTX-A and BTX-A + zinc groups. At 12 weeks, the nuclear factor kappa beta (NF-κB) mRNA expression was significantly higher in BTX-A + zinc and BTX-A groups compared to BTX-A + copper group and relative to the control group. ConclusionZinc supplementation significantly improved the effectiveness and durability of BTX-A, whereas copper supplementation reduced its efficacy.

Co-Stimulation with TWEAK and TGF-β1 Induces Steroid-Insensitive TSLP and CCL5 Production in BEAS-2B Human Bronchial Epithelial Cells.

Steroid-resistant asthma is a common cause of refractory asthma. Type 2 inflammation is the main inflammatory response in asthma, and the mechanism underlying the steroid-resistance of type 2 inflammation has not been completely elucidated. Tumor-necrosis-factor-like apoptosis-inducing factor (TWEAK) and transforming growth factor (TGF)-β1 are involved in epithelial-mesenchymal transition (EMT) and the production of thymic stromal lymphopoietin (TSLP) and C-C motif chemokine ligand 5 (CCL5). We herein hypothesize that the combined exposure to TWEAK and TGF-β1 may result in the development of steroid resistance in bronchial epithelial cells. The bronchial epithelial cell line BEAS-2B was cultured with or without TGF-β1 or TWEAK, in the presence or absence of dexamethasone (DEX). The roles of Smad-independent pathways and MAP kinase phosphatase 1 (MKP-1) were also explored. Co-stimulation of TWEAK and TGF-β1 induced E-cadherin reduction, N-cadherin upregulation, and TSLP and CCL5 production, which were not suppressed by DEX. Inhibition of the nuclear factor kappa beta (NF-κB) and mitogen-activated protein kinase pathways downregulated steroid-unresponsive TSLP and CCL5 production, whereas knockdown of MKP-1 improved steroid-unresponsive TSLP production, induced by co-stimulation with TWEAK and TGF-β1. Therefore, co-stimulation with TWEAK and TGF-β1 can induce the steroid-insensitive production of TSLP and CCL5 in the bronchial epithelium and may contribute to airway inflammation.

RANK-RANKL-OPG System in COVID-19: Examining the Pathway Among Healthcare Workers in Nigeria

<i>Background</i>: Coronavirus disease 2019 (COVID-19) influences bone metabolism by altering the RANK-RANKL-OPG system. However, this has not been validated, especially among Nigerians. Consequently, the current study explored the influence of the disease on this vital skeletal pathway among Nigerian healthcare workers (HCWs). <i>Methods</i>: This was a prospective longitudinal study conducted in the Department of Chemical Pathology of the Rivers State University Teaching Hospital among HCWs in Rivers State, Southern Nigeria. Eligible HCWs (n=76) with moderate RT-PCR-confirmed COVID-19 were recruited and compared with age and sex-matched healthy controls. Demographic, anthropometric, clinical, and laboratory data were obtained at baseline upon COVID-19 onset and followed up on days four and seven. Statistical analysis was done using descriptive/inferential statistics at a p-value <0.05. <i>Results:</i> The HCWs with moderate COVID-19 had higher serum levels of pro-inflammatory markers (IL-1β, IL-6, TNF-α) and receptor activator of nuclear factor kappa beta ligand (RANKL) but lower serum levels of osteoprotegerin (OPG) t COVID-19 diagnosis compared to the healthy controls (p<0.05). Among the HCWs with positive COVID-19 status, an increasing trend of these inflammatory markers and RANKL was observed from day one to day four and day seven, but a decreasing trend of OPG levels was observed (p<0.05). On day seven following COVID-19 diagnosis among the HCWs, a positive relationship was established between serum RANKL and all the pro-inflammatory markers (p<0.001) while an inverse relationship was only observed between OPG and IL-1β pro-inflammatory marker (p<0.05). <i>Conclusion:</i> The study findings corroborate the negative influence of COVID-019 on the RANK-RANKL-OPG system in favor of exaggerated osteoclastogenesis.

Proteomics Reveals Divergent Cardiac Inflammatory and Metabolic Responses After Inhalation of Ambient Particulate Matter With or Without Ozone.

Inhalation of ambient particulate matter (PM) and ozone (O3) has been associated with increased cardiovascular morbidity and mortality. However, the interactive effects of PM and O3 on cardiac dysfunction and disease have not been thoroughly examined, especially at a proteomic level. The purpose of this study was to identify and compare proteome changes in spontaneously hypertensive (SH) rats co-exposed to concentrated ambient particulates (CAPs) and O3, with a focus on investigating inflammatory and metabolic pathways, which are the two major ones implicated in the pathophysiology of cardiac dysfunction. For this, we measured and compared changes in expression status of 9 critical pro- and anti-inflammatory cytokines using multiplexed ELISA and 450 metabolic proteins involved in ATP production, oxidative phosphorylation, cytoskeletal organization, and stress response using two-dimensional electrophoresis (2-DE) and mass spectrometry (MS) in cardiac tissue of SH rats exposed to CAPs alone, O3 alone, and CAPs + O3. Proteomic expression profiling revealed that CAPs alone, O3 alone, and CAPs + O3 differentially altered protein expression patterns, and utilized divergent mechanisms to affect inflammatory and metabolic pathways and responses. Ingenuity Pathway Analysis (IPA) of the proteomic data demonstrated that the metabolic protein network centered by gap junction alpha-1 protein (GJA 1) was interconnected with the inflammatory cytokine network centered by nuclear factor kappa beta (NF-kB) potentially suggesting inflammation-induced alterations in metabolic pathways, or vice versa, collectively contributing to the development of cardiac dysfunction in response to CAPs and O3 exposure. These findings may enhance understanding of the pathophysiology of cardiac dysfunction induced by air pollution and provide testable hypotheses regarding mechanisms of action.

Assessment of IV albumin and ringer lactate on the acute oral toxicity of acetylsalicylic acid in albino rats

BackgroundDespite the frequent inclusion of fluid therapy in the treatment of many conditions, there are limited studies available to provide an evidence-based specific recommendation for fluid therapy in acute drug toxicity. Salicylate toxicity is considered one of the common clinical problems. It is commonly associated with fatal complications and even can lead to death. The study was designed to investigate the effects of various IV fluid types as isotonic saline (NaCl 0.9%), Ringer lactate (RL), and albumin and their impact on acetyl salicylic acid (ASA) toxicity outcome in a rat model of acute salicylate toxicity.Sixty male Albino rats were divided into 10 groups of 6 rats each. The first four groups were the control, saline, RL, and albumin groups. The fifth group received two doses of ASA solution orally, and the next five groups were treated with IV fluids as follows: saline-ASA, RL-ASA, albumin-ASA, RL + albumin-ASA, and saline + albumin-ASA. Upon completion of the study, spirometry, arterial blood gas analysis (ABG), and serum liver and kidney function tests were done on all groups. Furthermore, quantitative real-time polymerase chain reaction (PCR) was used to assess interleukin-6 (IL6), nuclear factor kappa beta (NF-kβ), and beta-actin mRNA gene expression of histopathology and immunohistochemistry assessments were also performed on liver and kidney tissues.ResultsThe results revealed the ASA group showed marked deterioration across all the investigated parameters. The groups that received saline and RL showed improvements in the following: respiratory rates, ABG, liver and kidney function, and histopathological findings.The RL + albumin group did not show any improvements. The albumin group and the saline + albumin group showed variable responses, ranging from mild improvement to no improvement.ConclusionsThe saline and RL groups showed positive results; however, the RL + albumin group showed the worst outcomes. The inclusion of albumin did not appear to provide any extra benefits and produced varying results.

Metformin: Beyond Type 2 Diabetes Mellitus.

Metformin was developed from an offshoot of Guanidine. It is known to be the first-line medication for type 2 diabetes mellitus, polycystic ovarian syndrome, and weight reduction. Metformin has also been shown to have effectiveness in the management of non-alcoholic fatty liver disease (NAFLD), liver cirrhosis, and various carcinomas like hepatocellular, colorectal, prostate, breast, urinary bladder, blood, melanoma, bone, skin, lung and so on. This narrative review focuses on the effect of metformin on non-alcoholic fatty liver disease, liver cirrhosis, and hepatocellular carcinoma. The search platforms for the topic were PubMed, Scopus, and Google search engine. Critical words for searching included 'Metformin,' AND 'Indications of Metformin,' AND 'Non-Alcoholic Fatty Liver Disease,' AND 'Metformin mechanism of action,' AND 'NAFLD management,' AND 'NAFLD and inflammation,' AND 'Metformin and insulin,' AND 'Metformin and inflammation,' AND 'Liver cirrhosis,' AND 'Hepatocellular carcinoma.' Lifestyle modification and the use of hypoglycemic agents can help improve liver conditions. Metformin has several mechanisms that enhance liver health, including reducing reactive oxygen species, nuclear factor kappa beta (NF-κB), liver enzymes, improving insulin sensitivity, and improving hepatic cell lipophagy. Long-term use of metformin may cause some adverse effects like lactic acidosis and gastrointestinal disturbance. Metformin long-term overdose may lead to a rise in hydrogen sulfide in liver cells, which calls for pharmacovigilance. Drug regulating authorities should provide approval for further research, and national and international guidelines need to be developed for liver diseases, perhaps with the inclusion of metformin as part of the management regime.

The modulatory effects of Lonicera caerulea L. extract and omega-3 on sarcopenia via regulating PI3K and FOXO signaling pathways in rats

Introduction: Sarcopenia is an inflammatory disease caused by a disruption of muscle homeostasis. Lonicera caerulea L. (Haskap berry) (HB) extract and omega-3 (ω−3) possess antioxidant and anti-inflammatory activities. This study assesses the potential ameliorating effects of HB extract and ω−3 supplementation on dexamethasone (DEXA)-induced sarcopenia. Methods: Rats were divided into five groups; the negative control group was injected with saline (i.p.); groups 2, 3, 4, and 5 were injected with DEXA (2 mg/kg/d, i.p.); groups 3 and 4 also received 400 mg/kg/d and 100 mg/kg/d of HB extract and ω−3, respectively, while group 5 received both treatments daily for 21 days. The ameliorative effects of treatments were investigated by measuring lysosomal proteolytic enzyme cathepsin activity, phosphatidylinositol 3 kinase (PI3K) activity, nuclear factor kappa beta (NF-κB) level, and heme oxygenase-1 (HO-1) activity. The gene expression levels of muscle ring-finger protein (MuRF) and forkhead box O (FOXO) transcription factor were also evaluated. Biochemical and histological examinations were done on muscle tissues. Results: DEXA caused a significant elevation (P<0.05) in NF-κβ level and cathepsin activity in muscle tissue. Also, it significantly upregulated the muscle atrophy markers. Meanwhile, PI3K and HO-1 activities were significantly reduced. HB extract and ω−3 administration significantly (P≤0.05) reversed these effects and downregulated the mRNA expression levels of MuRF and FOXO. Also, the histopathological examination confirmed these results. Conclusion: The current study proved the ameliorative effects of HB extract and ω−3 on sarcopenia parameters. Therefore, they might be potential therapeutic agents for myopathy/sarcopenia.

The regulatory effect and mechanism of traditional Chinese medicine on the renal inflammatory signal transduction pathways in diabetic kidney disease: A review.

Inflammatory injury is a critical factor in the occurrence and development of diabetic kidney disease (DKD). Signal transduction pathways such as the nuclear factor kappa beta (NF-κB), mitogen-activated protein kinase (MAPK), NOD-like receptor protein 3, and Smads are important mechanisms of inflammatory kidney injury in DKD, and the NF-κB pathway plays a key role. The inflammatory factor network formed after activation of the NF-κB pathway connects different signaling pathways and exacerbates renal inflammatory damage. Many traditional Chinese medicine compounds, single agents, effective components and active ingredients can regulate the expression of key molecules in the signaling pathways associated with inflammatory injury, such as transforming growth factor-β activated kinase 1, NF-κB, p38MAPK, NOD-like receptor protein 3, and Smad7. These treatments have the characteristics of multiple targets and have multiple and overlapping effects, which can treat DKD kidney inflammation and injury through multiple mechanisms and apply the "holistic concept" of traditional Chinese medicine.

Epithelial Interleukin-1 Receptor-Like-1 Activation Is Contingent on Interleukin-33 Isoforms and Asthma-Related Receptor Variation.

The interleukin-33/interleukin-1 receptor-like-1 (IL-33/IL1RL1) signalling pathway is implicated in asthma pathogenesis, with IL1RL1 nonsynonymous genetic polymorphisms associated with disease risk. We aimed to determine these variants' effect on IL1RL1 signalling induced by different IL33 isoforms thought to be elevated in the asthmatic airway. In a project funded by GSK plc, which has developed an IL-33 receptor inhibitor for asthma treatment, human embryonic kidney 293 (HEK293) cells expressing secreted embryonic alkaline phosphatase (SEAP) driven by a nuclear factor kappa-beta (NF-κB) promoter, were transiently transfected with IL1RL1, containing one of four extracellular and Toll/interleukin 1 receptor (TIR) domain haplotypes. Cells were stimulated with seven different splice and proteolytic-generated IL-33 isoforms (0.001-50 ng/mL) for 24 h. Supernatant SEAP activity and interleukin-8 (IL-8) levels were determined. Primary human bronchial epithelial cells (HBECs) representing different genotype carriers were stimulated with IL-33112-270 (50 ng/mL) and induced IL-8 mRNA expression measured. HEK293 cells carrying both asthma extracellular and TIR domain IL1RL1 risk haplotypes presented maximal IL33-driven signalling, with minimal signalling after IL-33 activation in other protective haplotypes. All IL-33 isoforms activated IL1RL1 but with differing magnitudes. Proteolytically cleaved IL3395-270 and IL33106-270 had the greatest effect and the IL33113-270, and Exon 3,4 deletion isoform exhibited the lowest. The effect of extracellular and TIR domain genetic variants on receptor signalling was replicated in primary HBECs. Maximal IL1RL1 signalling was observed in cells carrying both extracellular and TIR signalling domain risk haplotypes. Overall, our study suggests asthma patients carrying the extracellular and TIR domain risk haplotype and have a lung microenvironment that promotes elevated levels of cleaved IL33, particularly where IL3395-270 and IL33106-270 may be more amenable to IL33/IL1RL1 targeting.

The Effect of Mesenchymal Stem Cell Administration on the Expression of Nuclear Factor Kappa Beta, Neural Growth Factor Expression, and Prostaglandin E2 Concentration as Determinants of Pain in a Mouse Model of Endometriosis

Endometriosis is a significant reproductive health issue with pain being the primaru symptom. The complex pain in endometriosis involves the activation of macrophages, inflammatory activators such as NF-kB, neural growth factors (NGF), cytokines, adhesive molecules (ICAM-1), and mediators (PGE2) which they are stimulate sensory nerve fibers and produce nociceptive signals. This study is aim for investigate the effect of Mesenchymal Stem Cell (MSC) administration on the expression of NF-kB, NGF, and PGE2 concentration in a mouse model of endometriosis. Thirty-three mices were randomly divided into 3 groups consist non-endometriosis (P0), endometriosis without treatment (P1), and endometriosis with MSC administration (P2). The results showed MSC administration significantly reduced the expression of NF-kB by found scoring 2 groups (P1: 6.96 ± 0.13 and P2: 4.446 ± 0.228) and NGF (P1: 7.118 ± 0.629 and P2: 4.927 ± 1.187) also PGE2 (P1:1005.862 ± 176.656 and P2: 891.218 ± 54.031. In onclusion, this study demonstrates that MSC administration can able reduce the expression of NF-kB and NGF, but not to PGE2. The suggestion we can said is the MSC have potential therapeutic value in managing endometriosis-asssosciated pain by modulating inflammatory and neurotrophic factors. HIGHLIGHTS Pain in endometriosis, which is driven by inflammatory factors like NF-kB, NGF, cytokines, and PGE2, stimulating sensory nerve fibers and producing nociceptive signals. Investigate the effects of mesenchymal stem cell (MSC) administration on the expression of NF-kB, NGF, and PGE2 in a mouse model of endometriosis. Mesenchymal stem cell (MSC) administration was shown to reduce the expression of key inflammatory and neurotrophic factors—NF-kB and NGF—in a mouse model of endometriosis. Limited effect on reducing PGE2 concentration. These findings suggest that MSCs may have therapeutic potential for managing pain associated with endometriosis by modulating inflammatory and neurotrophic pathways. GRAPHICAL ABSTRACT

Exploring the role of macromolecular crowding and TNFR1 in cell volume control.

The excessive cosolute densities in the intracellular fluid create a physicochemical condition called macromolecular crowding (MMC). Intracellular MMC entropically maintains the biochemical thermodynamic equilibria by favoring associative reactions while hindering transport processes. Rapid cell volume shrinkage during extracellular hypertonicity elevates the MMC and disrupts the equilibria, potentially ushering cell death. Consequently, cells actively counter the hypertonic stress through regulatory volume increase (RVI) and restore the MMC homeostasis. Here, we establish fluorescence anisotropy of EGFP as a reliable tool for studying cellular MMC and explore the spatiotemporal dynamics of MMC during cell volume instabilities under multiple conditions. Our studies reveal that the actin cytoskeleton enforces spatially varying MMC levels inside adhered cells. Within cell populations, MMC is uncorrelated with nuclear DNA content but anti-correlated with the cell spread area. Although different cell lines have statistically similar MMC distributions, their responses to extracellular hypertonicity vary. The intensity of the extracellular hypertonicity determines a cell's ability for RVI, which correlates with nuclear factor kappa beta (NFkB) activation. Pharmacological inhibition and knockdown experiments reveal that tumor necrosis factor receptor 1 (TNFR1) initiates the hypertonicity-induced NFkB signaling and RVI. At severe hypertonicities, the elevated MMC amplifies cytoplasmic microviscosity and hinders receptor interacting protein kinase 1 (RIPK1) recruitment at the TNFR1 complex, incapacitating the TNFR1-NFkB signaling and consequently, RVI. Together, our studies unveil the involvement of TNFR1-NFkB signaling in modulating RVI and demonstrate the pivotal role of MMC in determining cellular osmoadaptability.

Inflammation and Apoptosis-Related Damage in Lung, Liver, and Kidney Tissues due to Subarachnoidal Hemorrhage

Objective: Oxidant and inflammatory substances released into the blood due to subarachnoidal hemorrhage (SAH) can pass into the peripheral compartment, causing distant organ damage due to blood-brain barrier permeability caused by oxidative stress, inflammation, and apoptosis. This study aimed to demonstrate the secondary damage to peripheral organs, including the lung, kidney, and liver, resulting from SAH. Material and Method: Twenty rats were divided into sham and SAH groups, each consisting of ten animals. In the SAH group animals, 0.3 mL autologous blood taken from the tail artery was injected into the cisterna magna for 2 minutes. Seven days after SAH formation, all animals were euthanized under anesthesia. Following decapitation, brain tissues, lung, liver, and kidney tissues were placed in 10% formaldehyde for histopathological and immunohistochemical analysis. Results: In the SAH group, neuronal degeneration in the cerebral cortex, and hyperemia and hemorrhage in the lung, kidney, and liver were observed histopathologically. In immunohistochemical examinations, decreased expression of brain-derived neurotrophic factor (BDNF) and neurofilament (NF) in the cerebral cortex, cerebellum, and hippocampus sections; In lung tissues, enhanced caspase (Cas)-3, hypoxia-inducible factor 1 alpha (Hif-1α) and nuclear factor kappa beta (NF-κB) expressions in the lung, Cas-5, cyclooxygenase-1 (Cox-1) and interleukin (IL)-1 expressions in the liver, Cas-3, Cox-1 and IL-3 expressions in the kidney were observed. Conclusion: Following SAH, in addition to damage to brain tissue, peripheral tissues such as the lung, kidney, and liver can also be damaged through inflammation and apoptosis.

Immune regulatory and anti-resorptive activities of tanshinone IIA sulfonate attenuates rheumatoid arthritis in mice.

Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation and painful joint destruction. Current treatments are helpful in RA remission, but strong immunosuppressive activity and patient resistance are clinical issues. This study explores a dual-action inhibitor, possessing both anti-inflammatory and anti-resorptive properties, as a novel treatment for RA. Therapeutic efficacy and mechanisms of ectosteric (tanshinone IIA sulfonate [T06]) and active site-directed (odanacatib [ODN]) inhibitors of cathepsin K (CatK) were evaluated in RA mouse models. Pathology was assessed through biochemical analyses and histopathological examination. Flow cytometry analysis was performed to characterize immune cells. Anti-inflammatory effects of T06 on nuclear factor kappa beta (NF-κB) pathway were studied in macrophages. T06 effectively lowered the number of joint-resident immune cells, accompanied by significantly reduced production of inflammatory cytokines and collagenolytic proteases. This also included the suppression of Th17 cells and IL-17, resulting in the reduction of osteoclasts in arthritic joints and amplification of the overall anti-resorptive effect of T06, which has been attributed to its selective inhibition of the collagenolytic activity of CatK by preventing its oligomerization. The anti-inflammatory mechanism of T06 was based on blocking the phosphorylation of IκBα in the NF-κB pathway, resulting in reduced activation and expression of inflammatory cytokines. In contrast, ODN had no effect on inflammation and disease progression and was limited to the inhibition of CatK. The combined anti-resorptive and anti-inflammatory activities characterize T06 as a novel therapeutic agent for RA.

7,8‑dihydroxyflavone reduces lipid peroxidation, proinflammatory cytokines, and mediators in chemically induced‑phenylketonuria model.

Phenylketonuria (PKU) stems from a rare genetic metabolic imbalance attributed to an insufficiency in the enzyme phenylalanine hydroxylase. Within the context of PKU, brain‑derived neurotrophic factor (BDNF) plays a pivotal role in brain function. 7,8‑dihydroxyflavone (7,8‑DHF) operates as a tropomyosin receptor kinase B (TrkB) agonist, mimicking the effects of BDNF. This study aimed to examine the effects of administering 7,8‑DHF in chemically‑induced rat models specifically induced to simulate PKU chemically. The rats were subcutaneously injected with phenylalanine and p‑chlorophenylalanine, a phenylalanine hydroxylase inhibitor, along with 7,8‑DHF. The injections began on the 2nd day after birth and continued until the 10th day. Levels of interleukin‑1β (IL‑1β), interleukin‑6 (IL‑6), interleukin‑33 (IL‑33), BDNF, malondialdehyde (MDA), monoamine oxidase (MAO), and superoxide dismutase (SOD) in the brain tissues were quantified using the enzyme‑linked immunosorbent assay (ELISA). Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) was performed to assess the gene expressions of inducible nitric oxide synthase (iNOS), nuclear factor kappa beta (NF‑κB), caspase‑3, nuclear factor erythroid 2‑related factor 2 (Nrf2), heme oxygenase‑1 (HO‑1), and BDNF. The results showed a decrease in mRNA levels of iNOS, IL‑1β, IL‑6, and lipid peroxidation in the group that received 7,8‑DHF. These results indicate that administering 7,8‑DHF has the potential to reduce brain damage in PKU by lowering proinflammatory cytokine levels and lipid peroxidation in PKU models. Thus, 7,8‑DHF, as a small molecule, might offer a promising adjunct therapeutic approach for PKU.