Immunopharmacology and Immunotoxicology最新文献

Objectives: Epilepsy is a chronic neurological condition with complex etiopathogenesis, treated with antiepileptics. In addition to their ability to regulate the activation threshold of neurons, antiepileptics have demonstrated a potential in shaping inflammation and the immune response. The main objective of our study was to analyze the effects of valproate, carbamazepine, and lamotrigine (commonly used antiepileptics) on viability, lymphocyte proliferation, and cytokine production by human peripheral blood mononuclear cells (PBMCs).

Methods: PBMCs were treated with different concentrations of antiepileptics, with or without phytohemagglutinin (PHA). Cytotoxicity, assessed by viability and apoptosis/necrosis assay, was determined by flow cytometry using the Annexin V/Propidium iodide (PI) staining method. Proliferation was determined using the MTT assay, whereas cytokine levels were assessed by the ELISA assay. A selective peroxisome proliferator-activated receptor gamma (PPAR-γ) antagonist (SR-202) was used to evaluate the involvement of PPAR-γ.

Results: Nontoxic concentrations of valproate and carbamazepine reduced the levels of three major proinflammatory cytokines (IL-1β, TNF-α, and IL-6) and impaired Th1 and Treg responses, without affecting the Th2 response. Lamotrigine did not exhibit immunomodulatory properties in this model. The effect of valproate on the production of proinflammatory and Th1 cytokines was significantly reversed by inhibiting PPAR-γ. In contrast, the blockade did not modify the effects of carbamazepine.

Conclusion: Our results demonstrated that valproate and carbamazepine, although similarly modulating the immune response in vitro, utilize different signaling mechanisms, in contrast to lamotrigine, which did not exhibit immunomodulatory effects.

Background: Zinc finger E-box binding homeobox 1 (ZEB1), a ferroptosis-associated gene, is upregulated in osteoarthritis (OA) articular cartilage. However, whether ZEB1 regulates ferroptosis in OA progression remain unclear.

Methods: ZEB1 protein levels in cartilage specimens from OA patients and normal controls were measured. Interleukin 1β (IL-1β)-induced chondrocyte injury model was established, followed by short hairpin RNA (shRNA)-mediated ZEB1 silencing in chondrocytes. Chondrocyte viability, apoptosis, inflammatory cytokine expression, extracellular matrix (ECM) degradation were assessed. Moreover, ferrous ion (Fe²⁺) level, reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase 4 (GPX4), mitochondrial membrane potential (MMP) and ATP level were determined. ZEB1-mediated transcriptional regulation of heat shock protein family A member 5 (HSPA5) was validated. Rescue experiments were conducted to validate the ZEB1/HSPA5 regulatory axis in chondrocyte injury. OA mouse model was constructed, and ZEB1 shRNA was injected into OA mice. The pathological changes in cartilage tissues were detected.

Results: ZEB1 was upregulated in OA cartilage tissues. ZEB1 silencing attenuated IL-1β-induced apoptosis, inflammation, and ECM degradation. IL-1β treatment increased Fe²⁺, ROS, and MDA levels and decreased GPX4 and GSH levels in chondrocytes, while ZEB1 silencing reversed these changes. ZEB1 silencing abrogated IL-1β-induced MMP and ATP reduction. Mechanistic studies revealed that ZEB1 transcriptionally inhibited HSPA5 expression in chondrocytes. HSPA5 silencing abrogated the protective effects of ZEB1 silencing. Additionally, ZEB1 silencing alleviated articular cartilage degradation, inflammatory response, and iron deposition.

Conclusion: ZEB1 silencing ameliorated IL-1β-induced chondrocyte injury and OA progression by suppressing ferroptosis and mitochondrial dysfunction via HSPA5 inhibition.

Background: Renal fibrosis is a hallmark feature of chronic kidney disease (CKD) and contributes to local inflammation and impaired renal function. The study aimed to investigate the effects of an active compound, alpha-cyperone, on renal fibrosis, inflammation, and oxidative stress in CKD mice.

Methods: After subtotal nephrectomy, mice were administered with alpha-cyperone (5, 10, and 20 mg/kg) or captopril (the positive control) daily via oral gavage. Masson trichrome staining and periodic Acid-Schiff staining were performed to measure fibrotic areas and renal histopathological changes. An enzyme-linked immunosorbent assay was conducted to measure inflammatory cytokine levels in mouse serum samples. Superoxide anion concentrations, reactive oxygen species (ROS) level, and superoxide dismutase (SOD) activity were analyzed. Immunohistochemistry was performed to detect inflammatory and oxidative stress markers (Ly6G and 8-OHdG) in mouse renal tissues. Protein levels of factors involved in NF-κB signaling and Akt/Nrf2/HO-1 signaling were quantified by western blotting.

Results: Alpha-cyperone at the dose of 10 mg/kg significantly improved renal functions in CKD model mice and alleviated tubulointerstitial fibrosis, with effects comparable to those of captopril. After CKD modeling, serum levels of inflammatory mediators, Ly6G levels, superoxide anion activity, ROS levels, and 8-OHdG levels were markedly increased, while the activity of antioxidant enzyme SOD was reduced. All these changes were ameliorated by alpha-cyperone or captopril treatment. Moreover, alpha-cyperone inhibited the NF-κB signaling and activated the Akt/Nrf2/HO-1 signaling in renal tissues of CKD mice.

Conclusion: Alpha-cyperone ameliorates renal fibrosis in CKD mice by inhibiting inflammation and oxidative stress via NF-κB and Akt/Nrf2/HO-1 pathways.

Background: The anesthetic sevoflurane can cause cognitive dysfunction and may be involved in mediating DeoxyriboNucleic Acid (DNA) methylation. In this study, we dig into the mechanism of sevoflurane inducing cognitive dysfunction via DNA methylation pathway.

Methods: In vivo and in vitro experiments were performed in sevoflurane-induced rat models and microglia. In vivo experiments included Morris water maze, Western blot, methylation analysis and immunofluorescence, while in vitro experiments consisted of quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. MK2206 was used as a protein kinase B (AKT) inhibitor.

Results: Sevoflurane induced cognitive dysfunction in rats, promoted levels of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), Interleukin (IL)-18, IL-1β, and ionized calcium-binding adapter molecule 1 (Iba-1) proteins, and inhibited Period2 (PER2) expression by enhancing methylation modification. PER2 was found to be located in microglia. Sevoflurane activated DNA methyltransferases (DNMTs) expression and suppressed PER2 in vitro. PER2 overexpression reduced NLRP3 inflammasomes-related protein expressions and restored AKT activation in sevoflurane-treated cells. Furthermore, MK2206 reversed the inhibitory effect of PER2 overexpression on cellular inflammation and AKT pathway activation.

Conclusion: Sevoflurane affects AKT pathway-suppressed NLRP3 inflammasomes in microglia by modulating PER2 methylation, thereby contributing to cognitive dysfunction.

Background: Ferroptosis-associated insults play a critical role in the pathological development of septic cardiomyopathy. Fenofibrate (Feno) is a fibrate drug used to treat high triglycerides and high cholesterol, however its pharmacological function in septic cardiomyopathy is not well understood.

Materials and methods: We allocated 36 male C57BL/6J mice into four groups (n = 9/group): Vehicle, Feno, LPS, and LPS+Feno. Techniques included hematoxylin-eosin (HE) staining, LDH assay, ELISA, echocardiography, measurement of MDA, GSH, Fe2⁺, real-time PCR, and western blot analysis.

Results: Administration of Feno significantly mitigated myocardial injury by reducing serum CK-MB levels from 963.8 U/L to 512.5 U/L, cTnI from 0.65 g/L to 0.36 g/L, and LDH from 552.4 U/L to 372.1 U/L. Feno improved cardiac function by increasing ejection fraction from 65.5% to 78.5% and fractional shortening from 42.3% to 57.3%. Feno also inhibited inflammatory cytokines IL-6 and TNF-α, reduced MDA levels, increased GSH levels, and restored GPX4, FTH1, and SLC7A11 expression. The protective effects of Feno may be associated with the YAP1 signaling pathway.

Conclusion: Our findings suggest that Feno has the potential to protect against LPS-induced cardiac injury through the alleviation of ferroptosis, offering a promising therapeutic strategy for septic cardiomyopathy. However, the study is limited by the use of a single animal model and the lack of translational data in humans.

Introduction: The immunosuppressant drug cyclosporine A (CsA) demonstrates anti-inflammatory properties in numerous pathological conditions. It acts through modulating T-cell receptor signaling, reducing the expression of inflammatory cytokines, and inhibiting mitochondrial permeability, besides modulating vascular response. These features make it a potential drug to prevent or treat septic acute kidney injury (AKI).

Objective: In this study, we investigated whether CsA exerts a protective effect against hemodynamic, inflammatory, and renovascular consequences of sepsis and whether these effects are modulated by adenosine receptor signaling.

Material and methods: Cecal ligation and puncture (CLP) was utilized to induce sepsis 24 h before hemodynamic and renovascular studies were implicated. Renal vasoconstrictions and vasodilatations were induced by cumulative bolus injections of phenylephrine (PE, 0.41-900 ng) and acetylcholine (ACh, 0.01-7.29 nmol), respectively.

Results: The data showed that CsA abrogated CLP-evoked hypotension, tachycardia, and impaired renovascular responsiveness. Similarly, the elevation in kidney biomarkers together with the pro-inflammatory cytokines (Tumor necrosis factor-alpha (TNFα) and Interleukin-6 (IL-6)) were also blunted after CsA administration. Likewise, the elevation in nuclear factor kappa-light-chain enhancer of activated B cells (NFκB) and decrease in A2BRs renal tubular expression in sepsis was reversed in CsA-treated rats. These advantageous effects of CsA disappeared upon concurrent exposure to the selective A2BR antagonist, Alloxazine.

Conclusion: These results suggest a key role for functional A2BR in CsA counteracting CLP-induced hemodynamic, inflammatory, and renal dysfunction in rats.

Background: Immune checkpoint inhibitors (ICIs) have shown significant advantages in the treatment of lung cancer. Several studies have reported immune-related adverse events (irAEs) induced by ICIs. However, in clinical practice, irAEs occasionally cause lymphadenopathy, which can be mistaken for tumor progression, making it more challenging to accurately assess the patient's condition.

Case presentation: This research report documents a rare clinical case of a patient with early-stage non-small cell lung cancer (NSCLC) who developed systemic lymphadenopathy during treatment with a PD-1 ICI following surgical resection. The patient developed widespread lymphadenopathy during postoperative PD-1 antibody maintenance therapy, accompanied by a series of irAEs, including persistent low cortisol levels, sluggish responses, memory loss, and stiffness in distal limbs and shoulder joints. Given the clinical presentation, the possibility of lymph node metastasis from lung cancer could not be entirely excluded. However, lymph node biopsy revealed reactive hyperplasia. After receiving corticosteroid treatment, the enlarged lymph nodes significantly reduced in size, and the associated low cortisol symptoms disappeared. During subsequent follow-up, the patient showed significant improvement and maintained a relatively stable condition.

Results: In cases of postoperative generalized lymphadenopathy in NSCLC patients, if tumor recurrence is suspected, careful consideration is needed, especially in the era of ICI therapy. Postoperative PD-1 antibody maintenance therapy may induce reactive lymphadenopathy, including mediastinal lymph nodes. It is hypothesized that PD-1 antibodies cause T-cell activation in the lymph nodes.

Introduction: Sepsis-induced acute kidney injury (AKI) is a major cause of mortality in critically ill patients. Inflammation, oxidative stress, and apoptosis are key contributors to sepsis-related renal damage. Dapagliflozin (DPG), an SGLT2 inhibitor, has demonstrated anti-inflammatory and nephroprotective effects. This study aimed to investigate the renoprotective effects of DPG in a lipopolysaccharide (LPS)-induced sepsis model, focusing on inflammation, oxidative stress, and apoptosis in the kidneys.

Methods: Thirty-two male Wistar albino rats were divided into four equal groups: (1) Control group, which received saline for 5 days; (2) LPS group, which received oral saline for 5 days and a single intraperitoneal LPS injection (5 mg/kg on day 5); (3) LPS+DPG group, which received oral DPG (10 mg/kg/day) for 5 days and a single intraperitoneal LPS injection (5 mg/kg on day 5); (4) DPG group, which received oral DPG (10 mg/kg/day) for 5 days. At the end of the experiment, samples were collected for histopathological, immunohistochemical, biochemical, and genetic analyses.

Results: DPG significantly reduced serum urea and creatinine levels, indicating improved renal function. Histopathological analysis of the kidney tissues revealed reduced inflammation, hemorrhage, and necrosis in the LPS+DPG group compared to the LPS group. Also, DPG lowered the expression of pro-inflammatory markers (APAF-1, TNF-α, VCAM-1), reduced oxidative stress (decreased OSI), and downregulated NLRP3, caspase-1, IL-1β, and IL-18 gene expression.

Conclusion: Prophylactic DPG administration exerts notable renoprotective effects in sepsis-induced AKI by mitigating inflammation, oxidative stress, and apoptosis. These findings highlight its potential as a preventive strategy, warranting further investigation in therapeutic contexts.

Purpose: Tofacitinib, a small molecule pan-JAK inhibitor, targets key inflammatory pathways that play a pivotal role in the pathophysiology of uveitis. Its ability to inhibit multiple cytokine signaling pathways makes it a promising candidate for the treatment of ocular inflammation. This study aimed to investigate the effect of oral tofacitinib on peptide-derived S-antigen (PDSAg)-induced chronic experimental autoimmune uveitis (EAU) in rats.

Materials and methods: Nineteen albino Wistar rats were divided into five groups. Groups 1-3 (5 rats each) were immunized with 15 micrograms PDSAg to induce EAU; Group 2 received tofacitinib (5 mg/kg) by gavage twice daily. Group 3 received saline in the same manner as Group 2. Group 4 (2 rats) was a healthy control group. Group 5 (2 rats) received only tofacitinib. Uveitis development and treatment efficacy were evaluated using clinical scoring based on the the signs of anterior segment inflammation and histological scoring based on the deterioration of the retinal architectural structure.

Results: Uveitis confirmed based on histological evidence was observed in all EAU groups (Groups 1-3) compared to the healthy control group (Group 4) (p < 0.001, Mann-Whitney U test). Tofacitinib significantly delayed the onset of uveitis in Group 2 when compared with Groups 1 and 3, which did not receive tofacitinib (p < 0.001, Mann-Whitney U test). Histological scores also showed a trend toward reduction (p = 0.393, Mann-Whitney U test).

Conclusions: Oral tofacitinib delayed uveitis onset and reduced clinical and histological findings, suggesting its potential as an alternative treatment for uveitis.