Immunopharmacology and Immunotoxicology最新文献

Objective: This study aimed to investigate the role of Kindlin-2 in HCC and its underlying molecular mechanisms, focusing on its regulation of the Fas/FasL signaling pathway.

Materials and methods: In vitro, Hep3B and HepG2 cells were treated with Kindlin-2 siRNA, a Fas activator, and a combination of Kindlin-2 siRNA and Fas siRNA. Cell proliferation, apoptosis, and cell cycle progression were evaluated using CCK-8 assays and flow cytometry, while the expression of associated proteins was analyzed through Western blotting. In vivo, a nude mouse xenograft model was established, and the expression levels of apoptosis and cell cycle proteins were assessed using Western blotting and immunohistochemistry.

Results: Silencing Kindlin-2 significantly upregulated the expression of Fas and Fas ligand (FasL), activating the Fas/FasL signaling pathway. This activation promoted the recruitment of FADD, leading to the activation of caspase-8 and caspase-3, inducing apoptosis and causing G1 phase cell cycle arrest.

Discussion and conclusion: This study revealed that Kindlin-2 inhibited apoptosis in HCC by negatively regulating the Fas/FasL signaling pathway. Kindlin-2 reduced apoptosis in HCC cells by suppressing the activation of the Fas/FasL pathway, thereby promoting tumor progression.

Introduction: In this study, our aim was to investigate the protective effects of myricetin (single dose-100 mg/kg) on CLP-induced rat sepsis model by analyzing some immune mechanisms including inflammation and oxidative stress by different techniques such as Immunohistochemistry, ELISA, tissue biochemistry and Western Blotting.

Methods: Twenty-eight Wistar albino rats were divided into 4 groups. The pro-inflammatory and anti-inflammatory cytokine levels were measured by ELISA technique. CD68 and Nuclear-Factor-Kappa-B (NF-κB) positivity rates were detected by IHC. Some of oxidative stress parameters were measured by tissue biochemistry, while Toll-like receptor-4 (TLR4) expression others were detected by Western blot technique.

Results: Sepsis caused a significant increase in all pro-inflammatory cytokine and oxidant levels. Also, it led to an increase in the positivity of CD68 and NF-κB markers as well as the expression levels of TNF-alpha, IL-1-beta, TLR4, Keap-1. However, single dose myricetin application normalized pro-inflammatory cytokine levels, increased anti-oxidant and anti-inflammatory cytokine levels, decreased positivity of CD68 and NF-κB and increased NRF2 and HO-1 expressions.

Discussion: As a conclusion, the beneficial effect of myricetin on lung injury also involved inhibition of TLR4/NF-κB pathway, suppression of proinflammatory cytokines and induction of anti-inflammatory cytokine production, regulation of oxidant and anti-oxidant system parameters, and activating the NRF2/Keap1/HO-1 pathway.

Objective: This study aimed to investigate the possible protective effect of roflumilast (RFL) on cyclophosphamide (CP)-induced ovarian toxicity as well as the possible underlying mechanism.

Material and methods: Female Wistar rats received the vehicle (n = 6) or CP (200 mg/kg, i.p.). The other 2 groups (n = 6 for each) were orally pretreated with RFL at dosages of 0.5 and 1 mg/kg, respectively, for 14 days and then after one hour of RFL administration on the 14th day, rats were intraperitoneally administered a single dose of CP. Serum and tissue samples were collected. Biochemical, real-time polymerase chain reaction, histopathological and immunohistopathological examination were carried out.

Results: RFL significantly elevated serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH) compared to the CP group. RFL remarkably elevated ovarian contents of Sirtuin-1 (SIRT1), heme oxygenase-1 (HO-1), and reduced nuclear factor-kappa B (NF-ĸb) p65/NF-ĸB ratio as compared to control CP group. Compared to the CP group, RFL significantly elevated Nrf2 gene expression, reduced malondialdehyde (MDA), and elevated the reduced glutathione (GSH) ovarian content. It also reduced the protein expression of TNF-α and caspase-3.

Conclusion: It can be concluded that RFL (0.5 and 1 mg/kg) protected rats against CP-induced ovarian toxicity via altering the SIRT1/Nrf2/NF-ĸB pathway, ameliorating histopathological changes in addition to its anti-apoptotic effect.

Background: Nintedanib is a potent antifibrotic angiokinase inhibitor approved for various fibrotic lung diseases. Potential therapeutic efficacy of nintedanib in various inflammatory diseases is under investigation. In this study, we investigated the therapeutic effect of nintedanib on adipogenesis and fibrosis in orbital fibroblasts in patients with Graves' orbitopathy (GO).

Methods: Primary orbital fibroblasts were cultured from orbital connective tissue of patients with GO and healthy controls. The cells were pretreated with nintedanib before stimulation with either interleukin (IL)-1β, transforming growth factor (TGF)-β, insulin-like growth factor-1, or IL-11. Fibrosis-related and intracellular signaling protein expressions were assessed using western blotting. Hyaluronan and procollagen concentrations were quantified using enzyme-linked immunosorbent assay. Adipogenesis was quantified by Oil Red O staining and the levels of adipogenic transcription factors were determined by Western blot.

Results: TGF-β-induced fibronectin and collagen 1/3 protein expression was abrogated by nintedanib treatment. Nintedanib decreased the phosphorylation of signal transducer and activator of transcription 3, SMAD 2/3, Akt, c-Jun N-terminal kinase, and extracellular regulated protein kinase. Exposure to nintedanib hindered adipocyte differentiation and expression of adipogenic transcription factors, including peroxisome proliferator-activated receptor γ, CCAAT/enhancer-binding protein α/β, adipocyte protein 2, adiponectin, and leptin. Additionally, nintedanib reduced hyaluronan and procollagen secretion.

Conclusions: Nintedanib suppressed profibrotic protein production, adipogenesis, and hyaluronan production in in vitro. These findings indicate the potential therapeutic efficacy of nintedanib in GO management.

Background: Inflammatory bowel disease (IBD) is a chronic relapsing gastrointestinal disorder. Infliximab (INF) has shown good efficacy in IBD treatment, but its specific impact requires further exploration. This study aimed to assess the effects of intravenous INF on clinical symptom scores and serum cytokine levels in IBD patients.

Methods: A retrospective review of 126 IBD patients treated with INF was conducted. Baseline data, Mayo scores, Crohn's Disease Activity Index (CDAI) scores at 6 and 12 months, and serum levels of TNF-α, IL-6, IL-10, and CRP were recorded. Correlations between disease activity scores and inflammatory markers were analyzed, and the relationship between baseline indicators and treatment efficacy was examined.

Results: At 12 months, Mayo and CDAI scores, TNF-α, IL-6, and CRP levels were significantly reduced, while IL-10 levels increased. Disease activity scores positively correlated with TNF-α, IL-6, and IL-1β, and negatively with IL-10. Factors such as Crohn's disease subtype, age, high baseline CDAI or Mayo scores, elevated TNF-α, IL-6, CRP, and longer disease duration were associated with poorer outcomes (p < 0.05). Multivariate analysis identified disease type, high baseline disease activity, long disease duration, and elevated inflammatory markers as independent risk factors. Adverse reactions were infrequent, with no serious adverse events reported.

Conclusion: Intravenous INF effectively improves clinical symptoms and modulates inflammatory cytokines in IBD patients, with favorable safety and increasing efficacy over time. However, the limited sample size and lack of long-term data warrant further validation in larger, prospective multicenter studies.

Objectives: Periodontal disease is a chronic inflammatory disease caused by periodontopathogenic bacteria, and its progression leads to periodontal tissue destruction and tooth loss. Zerumbone is a bioactive substance found in ginger (Zingiber zerumbet) and is known to have bioactive effects such as anticancer effects, but there have been no attempts to use it for periodontitis treatment. In addition, there have been no reports examining its effects on periodontal tissue component cells. In this experiment, we aimed to determine whether zerumbone affects the production of inflammatory mediators induced by tumor necrosis factor (TNF)-α in human periodontal ligament cells (HPDLCs), including its effects on signaling pathways.

Methods: HPDLCs were stimulated by TNF-α (10 ng/ml) with or without zerumbone (6.25, 12.5, or 25 µM). Cytokine production in supernatant was determined using ELISA. Activation of signal transduction pathways and intracellular protein expression were investigated using the western blot analysis.

Results: Zerumbone significantly suppressed TNF-α-induced production of CC chemokine ligand 2 (CCL2), CCL20, CXC chemokine ligand 10 (CXCL10), and interleukin-6 (IL-6) in HPDLCs. In addition, zerumbone decreased intercellular adhesion molecule-1 (ICAM-1) and cyclooxygenase-2 (COX-2) expression in TNF-α-stimulated HPDLCs. Furthermore, zerumbone suppressed activation of nuclear factor (NF)-κB and signal transducer and activator of transcription 3 (STAT3) pathways in TNF-α-treated HPDLCs. Finally, zerumbone enhanced the production of heme oxygenase-1 (HO-1), an antioxidant enzyme, in HPDLCs.

Conclusion: These results suggest that zerumbone suppressed the production of several inflammatory mediators by inhibiting the NF-κB and STAT3 pathways in HPDLCs.

Objectives: Juvenile idiopathic arthritis (JIA) is the most common rheumatic disease in children. Nonsteroidal anti-inflammatory drugs (NSAIDs) and intra-articular corticosteroid injections are first-line therapy for oligoarticular JIA. NSAIDs Adverse events (AEs) include gastrointestinal ulcers/bleeding and impaired renal function. The most prescribed NSAIDs for oligoarticular JIA are ibuprofen and naproxen. However, direct comparison between these drugs is lacking. We aimed to compare the efficacy and safety of ibuprofen versus naproxen for oligoarticular JIA.

Methods: This is a bi-national retrospective study of oligoarticular JIA patients treated with either ibuprofen or naproxen as first-line therapy. Efficacy was defined as patients that achieved complete response (no evidence for arthritis). Safety was assessed by the occurrence of adverse events during follow-up.

Results: Of 164 patients, 103 were treated in the Israeli group and 61 in the US group. The study population had a mean age of 4.49 ± 3.55 years, with F:M ratio of ∼2.5:1. No significant difference was found in drug efficacy [Complete response was observed in 15% of the ibuprofen group vs. 17.3% in naproxen group (p = 0.7)]. Treatment duration > 28 days was associated with significantly higher odds for complete response (p = 0.021). For safety, 12 AEs were associated with naproxen, whereas no AEs were associated with ibuprofen (p = 0.004). Treatment was discontinued in all AEs cases.

Conclusions: Ibuprofen and naproxen showed similar albeit low efficacy which emphasizes their role as bridging therapy until IACI is achieved. However, ibuprofen showed better safety profile naproxen and therefore should be considered as first-line therapy.

Objective: Aim of this study is to compare the thiol/disulfide variables before treatment, at the 3^rd and 6^th months of biologic treatment in patients with axSpA.

Materials & methods: Consecutive patients with axial spondyloarthritis to whom biologic treatment was initiated in our clinic were enrolled upon consent. Demographics, clinical characteristics, laboratory parameters and treatment agents were collected. Disease activity scores and thiol-disulfide balance parameters were recorded at baseline and 3^rd, 6^th months of treatment. Statistical analyses were performed in all patients and in subgroups of ankylosing spondylitis and non-radiographic axial spondyloarthritis patients.

Results: In all patients, total thiol levels were significantly increased at 6^th month in comparison to baseline values (470.5 ± 74.7 vs 491.9 ± 69.6, p = 0.047). Native thiol levels were increased at 6^th month close to significance (438.9 ± 70.4 vs 458.8 ± 63.7, p = 0.060). Moderately strong negative correlations were observed between native thiol levels and disease activity parameters (BASDAI: p = 0,019; ASDAS-CRP: p = 0,035; ASDAS-ESR: p = 0,030), and between total thiol levels and disease activity parameters (BASDAI: p = 0,031; ASDAS-CRP: p = 0,020; ASDAS-ESR: p = 0,026) at 6^th month evaluation.

Discussion & conclusions: Our results demonstrated oxidative stress reducing effect of biologics in axSpA patients parallel to suppression of disease activity at 6^th month of the treatment.

Objectives: Chitosan is widely used in medicine to regulate immune responses in T cells and dendritic cells. However, research on the regulation of mast cells (MCs) is scarce. Mas-related G-protein-coupled receptor X2 (MRGPRX2) is a key receptor that mediates MC activation. However, the inhibitory effects of chitosan on MRGPRX2 activation have not yet been reported. The aim of this study was to determine whether chitosan inhibits MRGPRX2-mediated MC activation and the molecular weight of chitosan with the best inhibitory effect.

Methods: Cytotoxic and activating effects of chitosan on LAD2 cells were evaluated in vitro. An in vitro MC degranulation reaction model and in vivo C48/80-induced local passive anaphylaxis mouse model were used to evaluate the inhibitory effect of chitosan on MRGPRX2 activation.

Key findings: Chitosan inhibited MC degranulation mediated by MRGPRX2 in vitro and reduced histamine, β-hexosaminidase, and cytokine release. Chitosan inhibited local pseudo-allergy and inflammatory mediator release by inhibiting MRGPRX2-mediated MC activation. Moreover, low-molecular-weight chitosan exhibited superior inhibitory activity against MRGPRX2.

Conclusions: Chitosan inhibited MRGPRX2-mediated MC degranulation in vivo and in vitro. Low molecular weight chitosan has the potential to be developed as a functional drug or food to assist in the treatment of MRGPRX2-regulated diseases.

Objective: Chemoresistance in gastric cancer poses a major challenge in treatment, necessitating the development of novel therapeutic strategies. This study evaluates the efficacy of ruxolitinib, a JAK1/2 inhibitor, in both sensitive and resistant gastric cancer cell lines.

Materials and methods: Gastric cancer cell lines, including sensitive (N87 and AGS) and resistant (N87-R and AGS-R) variants, were treated with ruxolitinib alone or in combination with chemotherapeutic agents. Apoptosis induction, mitochondrial function, oxidative stress, and key signaling pathways were analyzed. Tumor growth, p-STAT3 levels, and overall survival were evaluated in xenograft models.

Results: Ruxolitinib induced dose-dependent mitochondrial-mediated apoptosis in resistant cells and enhanced cytotoxicity in combination with chemotherapy in sensitive cells. The treatment inhibited phosphorylation of STAT3, Akt, and mTOR. Additionally, ruxolitinib reduced basal and maximal respiration rates while increasing ROS levels, suggesting mitochondrial dysfunction and oxidative stress. Resistant cells exhibited increased mitochondrial DNA content, elevated respiration rates, and higher ROS levels compared to sensitive cells, indicating alterations in mitochondrial biogenesis and redox homeostasis. These findings were supported by changes in gene expression related to mitochondrial function. In vivo, ruxolitinib significantly inhibited tumor growth and reduced p-STAT3 levels in resistant gastric cancer xenografts without causing significant weight loss. Furthermore, ruxolitinib treatment significantly prolonged overall survival in mice.

Conclusion: Ruxolitinib demonstrates potential in overcoming chemoresistance in gastric cancer by targeting mitochondrial function, oxidative stress, and key survival pathways. These findings support further investigation into its clinical application as an adjunct therapy for chemoresistant gastric cancer.