GPX4 Recombinant Rabbit Monoclonal Antibody

The immunosuppressive tumor microenvironment (ITME) promotes immune evasion and resistance to checkpoint blockade therapy. STING pathway activation offers a promising strategy to remodel the ITME, but existing agonists face limitations such as rapid degradation and poor bioavailability. Here, we developed natural compound rhein-based multifunctional bimetallic nanosheets (FGR NSs) composed of rhein (Rh), gadolinium (Gd 3+ ), and iron (Fe 3+ ) through a one-step symbiotic method to activate STING signaling and induce ferroptosis simultaneously. Upon decomposition in the tumor microenvironment, the natural product Rh triggers DNA double-strand breaks (dsDNA), promoting STING activation and increasing IFN-β secretion by 3.06-fold, while Fe 3+ drives ferroptosis through the Fenton reaction by catalyzing the conversion of endogenous hydrogen peroxide into highly toxic hydroxyl radicals and depleting glutathione (GSH), thereby promoting the generation of reactive oxygen species (ROS). Those mechanisms initiate both innate and adaptive immune responses and release abundant damage-associated molecular patterns (DAMPs, including CRT, ATP, HMGB1, and IFN-β) that promote dendritic cell maturation, CD8 + T-cell priming, and M2-to-M1 macrophage repolarization. In parallel, it suppresses the infiltration of immunosuppressive cells, including regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs). In vivo, FGR NSs elicit potent antitumor effects with 89.01 % tumor growth inhibition rates (TGI) in the subcutaneous breast cancer model. When combined with immune checkpoint inhibitor (αPD-1), a significant tumor inhibition in the lung metastatic model was achieved. Moreover, the Gd/Fe components enable T 1 /T 2 dual-mode MRI, positioning FGR as a promising theranostic platform for cancer immunotherapy.

Background Oligoasthenospermia is emerging as a critical cause of male infertility resulting from spermatogenesis dysfunction (SGD). Guilu Erxian glue (GLEXG) has traditionally been used to improve sperm quality, but its mechanism remains unclear. Purpose This study investigates the therapeutic mechanism of GLEXG and its active ingredient quercetin in a Tripterygium wilfordii polyglycoside (GTW)-induced SGD mouse model and GC-1 spermatogonial cells. Methods An SGD model was established by administering GTW (60 mg/kg/day, oral gavage) to BALB/c mice for four weeks, followed by treatment with GLEXG (2.25, 4.50, or 9.00 g/kg/day) or vitamin E (0.02 g/kg/day) for another four weeks. Therapeutic effects were assessed by sperm parameters. Key bioactive constituents and mechanistic pathways were identified using integrated network pharmacology and metabolomics analyses. The role of ferroptosis and associated signaling pathways was validated. Results GLEXG restored sperm motility to 78% and sperm concentration to 67% of normal levels ( p < 0.05) in SGD mice and improved testicular histopathology. Metabolomics indicated protection against ferroptosis through modulation of glutathione metabolism. Quercetin was identified as the key component targeting HIF-1α. In erastin-induced ferroptosis, GLEXG-containing serum and quercetin restored GC-1 cell viability by 60% and 46%, respectively; reduced lactate dehydrogenase release (76%; 50%), reactive oxygen species (ROS) (67%; 53%), malondialdehyde (MDA) (72%; 54%), and Fe²⁺ (96%; 86%); elevated the glutathione/glutathione disulfide (GSH/GSSG) ratio (78%; 54%); downregulated hypoxia-inducible factor-1alpha (HIF-1α) (64%; 45%); and upregulated glutathione peroxidase 4 (GPX4) (63%; 32%) and solute carrier family 7 member 11 (SLC7A11) (64%; 42%) ( p < 0.05). These effects were reversed by HIF-1α overexpression. In vivo , HIF-1α overexpression abrogated quercetin’s protection on sperm motility (53%), sperm concentration (53%), and testicular lesions ( p < 0.05). Conclusion GLEXG and quercetin alleviate GTW-induced SGD by inhibiting ferroptosis via HIF-1α/SLC7A11.

Background Diacylglycerol O-acyltransferase 1 (DGAT1) is crucial for triglyceride synthesis, yet its role in ischemic stroke remains unclear. This study investigated DGAT1 in ischemic stroke using middle cerebral artery occlusion (MCAO) rat models and highly differentiated PC12 cells subjected to oxygen–glucose deprivation/reoxygenation (OGD/R).Methods The therapeutic effects of DGAT1 inhibition in MCAO rats were assessed using the Zea-Longa score and 2,3,5-Triphenyltetrazolium chloride (TTC) staining. The effects on highly differentiated PC12 cells subjected to OGD/R were evaluated using the Cell Counting Kit-8 (CCK-8) and lactate dehydrogenase (LDH) assays. Ferroptosis-related mitochondrial damage was evaluated using transmission electron microscope. Additionally, the mechanisms by which DGAT1 inhibition regulates ferroptosis were further explored via immunohistochemistry, immunofluorescence, Western blotting, qPCR, JC-1 assay, and reactive oxygen species (ROS) detection.Results DGAT1 expression was elevated in both MCAO and OGD/R models. The DGAT1 inhibitor A 922500 improved neurological deficits, reduced infarct volume, and minimized neuronal loss in MCAO rats, while also enhancing cell viability and reducing LDH levels in OGD/R-treated PC12 cells. DGAT1 inhibition significantly alleviated ferroptosis in MCAO rats, as indicated by (i) reduced mitochondrial shortening and cristae disruption, (ii) decreased 4-HNE levels, (iii) reduced MDA and increased SOD, and (iv) lowered levels of inflammatory factors (IL-6, MCP-1, and TNF-α). Moreover, both in vivo and in vitro experiments showed that DGAT1 inhibition significantly increased Gpx4 levels, whereas lentiviral delivery of Gpx4 shRNA markedly reversed its beneficial effects. In MCAO rats, Gpx4 shRNA significantly elevated 4-HNE levels and exacerbated ferroptosis-related mitochondrial damage. In vitro, DGAT1 inhibition increased mitochondrial membrane potential and reduced ROS, whereas rotenone, a mitochondrial function inhibitor, decreased Gpx4 and impaired cell viability. Furthermore, DGAT1 inhibition significantly upregulated the key β-oxidation gene Cpt1a, whereas etomoxir, a β-oxidation inhibitor, reduced cell viability and mitochondrial membrane potential, increased ROS, and downregulated Gpx4.Conclusion sOur study suggests that DGAT1 inhibition may enhance β-oxidation and mitochondrial function, thereby increasing Gpx4 levels, suppressing ferroptosis, and ultimately exerting neuroprotective effects in ischemic stroke.

Background Radiation enteritis (RE) is a common complication in patients undergoing abdominal and pelvic radiotherapy. Despite the advancements in radiotherapy, effective treatments remain limited. WGX50, a bioactive compound from Sichuan pepper, has shown anti-inflammatory and antioxidant properties. This study investigates the protective effects of WGX50 on RE, focusing on its potential to reduce radiation-induced damage in the intestine. Methods Network pharmacology and molecular docking were used to identify the molecular targets of WGX50. In vitro, human intestinal epithelial cells (HIEC6) and colon cells (NCM460) were exposed to radiation and treated with WGX50. In vivo, C57BL/6 mice were administered WGX50 prior to radiation exposure. Various assays, including CCK-8, colony formation, flow cytometry, histopathology, and 16S rRNA sequencing, were performed to evaluate cell proliferation, apoptosis, oxidative stress, intestinal damage, and gut microbiota composition. Tissue transcriptome sequencing was conducted to explore differentially expressed genes. Results In vitro, WGX50 significantly mitigated radiation-induced cell damage, enhanced cell proliferation, and reduced apoptosis at non-toxic concentrations. In vivo, WGX50 treatment preserved intestinal morphology and reduced inflammatory infiltration in irradiated mice. WGX50 also protected goblet cells, maintaining mucin production and epithelial barrier function critical for intestinal homeostasis. Molecular docking, dynamics simulations and surface plasmon resonance (SPR) revealed stable binding of WGX50 to Epidermal Growth Factor Receptor (EGFR), key targets involved in oxidative stress regulation and ferroptosis inhibition. Mechanistically, WGX50 upregulated the EGFR-SLC7A11-GPX4 axis, suppressing ferroptosis and protecting intestinal cells. Additionally, 16S rRNA sequencing showed that WGX50 mitigated radiation-induced gut microbiota dysbiosis, preserving microbial diversity and promoting beneficial bacterial populations. Conclusion WGX50 demonstrates potent radioprotective effects by reducing oxidative stress, suppressing ferroptosis, and maintaining intestinal homeostasis, including goblet cell function and gut microbiota composition. These findings support WGX50’s potential as a novel therapeutic agent for the prevention and treatment of radiation enteritis.

Objective Caspase-6 is an important regulatory factor in innate immunity, inflammasome activation, and host defense, but its role in preeclampsia (PE) is unknown. This study aims to investigate the mechanism of Caspase-6 in the interaction between PE rats and macrophage-trophoblast cells, in order to provide a new theoretical basis for the treatment of PE. Methods Co-cultures of THP-1 cells and HTR8/SVneo cells were employed to investigate the HMGB1 signaling in macrophages (transfection with si-Caspase-6) and HTR8/SVneo cells. The PE rat model was constructed by using the reduced uterine perfusion pressure (RUPP) surgery to explore the therapeutic effects of bone marrow-derived macrophages (BMDM) transfected with si-Caspase-6 in PE rats. ELISA, Western blot, immunofluorescence, etc., were employed to characterize the expression of ferroptosis-related markers. Results Caspase-6 expression was significantly increased in CD14 + macrophages in the placental tissue of PE rats. Overexpression of Caspase-6 in THP-1 cells induced ferroptosis of HTR8/SVneo cells, but this process was blocked by anti-HMGB1 neutralizing antibody. Knockdown of Caspase-6 in macrophages could alleviate ferroptosis of HTR8/SVneo cells and restore its basic characteristics. Knockdown of Caspase-6 in BMDM downregulated ferroptosis in placental tissue of PE rats through HMGB1, thereby improving the disease phenotype in rats. Conclusion Knocking down Caspase-6 in BMDM regulated the crosstalk between macrophages and HTR8/SVneo cells through HMGB1, inhibiting HTR8/SVneo cell ferroptosis, thereby improving adverse pregnancy outcomes of PE.

A total of 22 terpenoids including one new triterpenoid ( 1 ), two new diterpenoids ( 2 and 3 ), and 19 known compounds ( 4 – 22 ) were isolated from the roots of Tripterygium regelii . Their structures were identified using NMR and HRESIMS techniques. Antiproliferative activity screening assays revealed that the new compound 2 , along with several known compounds, exhibits potential anti-colorectal cancer efficacy. Further investigations employing flow cytometry, immunofluorescence staining and electron microscopy demonstrated that compound 2 induces the generation of reactive oxygen species (ROS). This, in turn, promotes lipid peroxidation and mitochondrial damage, ultimately enhancing ferroptosis in colorectal cancer cells. These findings underscore the value of compound 2 as a potential candidate for anti-tumor drug development.

Objective Dipeptidyl peptidase-4 (DPP4)-targeted therapy is widely employed in the therapy of pulmonary diseases, but the role of its H3K4me1 modification in pulmonary arterial hypertension (PAH) disease remains unknown. This study aims to investigate the function of histone methyltransferase SET domain containing 7 (Set7)-mediated monomethylation of histone 3 lysine 4 (H3K4me1) modification of DPP4 in PAH, with the goal of providing new insights for the broader application of DPP4-targeted therapies. Methods The PAH mouse model was constructed and intervened with overexpression (oe) or knockdown (sh) of DPP4, sh-Set7, oe-NADPH oxidase 4 (NOX4) or sh-Set7 + erastin. Human pulmonary arterial endothelial cells were induced by hypoxia and treated with sh-DPP4, erastin, oe-DPP4, sh-Set7, oe-NOX4, oe-Set7 or oe-Set7 + ferrostatin-1. The enrichment of H3K4me1 level in the DPP4 promoter region was analyzed by ChIP and dual-luciferase assay. Pulmonary vascular remodeling, fibrosis, and endothelial injury were observed by echocardiography, HE, MASSON, and α-SMA staining. Ferroptosis markers and protein expression were measured using biochemical assay kits, RT-qPCR, WB, immunofluorescence, and transmission electron microscopy. Results Silencing DPP4 alleviates pulmonary vascular remodeling, fibrosis, and endothelial injury in PAH mice, reduces cardiac fibrosis and pulmonary inflammation, while improving mitochondrial damage in the lungs and downregulating the level of ferroptosis-related proteins. ChIP assays confirmed increased enrichment of H3K4me1 in the DPP4 promoter region in both hypoxia-induced endothelial cells and lung tissues of PAH mice. Overexpression of Set7 resulted in elevated H3K4me1 enrichment in the DPP4 promoter region and increased NOX4 protein expression. Ferrostatin-1 inhibited the promotion of oe-Set7 in hypoxia-induced endothelial cell injury. Silencing Set7 mitigated hypoxia-induced endothelial cell injury, ferroptosis, and inflammatory responses by downregulating DPP4/NOX4. Erastin reversed the treatment effect of sh-Set7 in PAH mice. Furthermore, Set7 knockdown ameliorated PAH in mice by suppressing DPP4/NOX4-mediated ferroptosis. Conclusion The H3K4me1 modification of DPP4 is upregulated in PAH, a process regulated by Set7. Silencing Set7 alleviates PAH by suppressing ferroptosis through the DPP4/NOX4 signaling pathway, offering a novel gene therapy approach for this disease.

Hyperlipidemic pancreatitis (HP) is an inflammatory injury of the pancreas triggered by elevated serum triglyceride (TG) levels. The mechanistic target of rapamycin (mTOR) signaling pathway plays a crucial role in regulating lipid homeostasis and inflammation. This study aimed to investigate whether the activity of mTOR complex 2 (mTORC2) affects the progression of HP and its underlying mechanisms. In vivo, a high-fat diet and retrograde administration of sodium taurocholate were employed to establish the HP models in rats, with pancreatic tissue pathology evaluated. The expression of Rictor and peroxisome proliferator-activator receptor (PPAR) was examined. The serum levels of TG, fatty acid metabolites, inflammatory and lipid metabolism-related factors were determined. In vitro, pancreatic acinar cells (PACs) were exposed to palmitic acid and cholecystokinin-8. PAC apoptosis, pyroptosis, and ferroptosis were assessed. In the HP models, rats and PACs exhibited upregulated Rictor and downregulated PPARα, and Rictor knockdown promoted PPARα expression. In vivo, Rictor knockdown decreased the serum levels of TG, α-amylase, total cholesterol, low-density lipoprotein cholesterol, lactate dehydrogenase, and inflammatory factors, while increasing high-density lipoprotein cholesterol levels. Rictor knockdown increased ACOX1 and CPT1α and decreased SREBP-1, CD36, SCD1, ACLY, and ACACA. Rictor knockdown reduced damage to pancreatic tissue structure. In vitro, Rictor knockdown inhibited PAC apoptosis, pyroptosis, and ferroptosis. Treatment with the PPARα antagonist GW6471 abolished the beneficial effects of Rictor knockdown. Rictor/mTORC2 deficiency reduces serum TG levels, maintains lipid homeostasis, and suppresses inflammation by inhibiting PPARα expression. Weakening mTORC2 activity holds promise as a novel therapeutic strategy for HP.

Background Ferroptosis plays a key role in the development of chronic obstructive pulmonary disease (COPD). Whether ginsenoside Rg1 improves cigarette smoke-induced COPD or whether ginsenoside Rg1 improves COPD by inhibiting ferroptosis remains unknown. Methods BEAS-2B cells were exposed to cigarette solution (CSE) for 24 hours and treated with ginsenoside Rg1, the ferroptosis inhibitor Fer-1, and the PERK inhibitor GSK. Cell viability, endoplasmic reticulum stress, mitochondrial morphology, membrane potential, reactive oxygen species (ROS), iron levels, and the expression of related proteins were detected using corresponding methods. A COPD mouse model was constructed using cigarette smoke (CS). Ginsenoside Rg1 and GSK were administered via tube feeding 15 days after successful modeling. Mouse lung tissues were evaluated by HE staining. The expression of inflammatory markers, ROS, iron content, and related proteins was detected using corresponding methods. Results The results demonstrated that in the CSE-exposed BEAS-2B cell model and CS-induced mouse COPD model, the expression levels of endoplasmic reticulum stress (ERS)-related factors such as GRP78 were increased, while those of the antioxidant markers GPX4 and GSH were significantly decreased. Ginsenoside Rg1 improved emphysema and inflammation by inhibiting ferroptosis in vivo and in vitro. Using a PERK inhibitor, we found that ginsenoside Rg1 inhibited ferroptosis in vivo and in vitro by regulating ERS. Conclusion This study showed that ginsenoside Rg1 alleviates cigarette smoke-induced COPD by regulating the PERK/ATF4 axis to inhibit ERS and ferroptosis.