Tucum-do-Cerrado (Bactris setosa Mart.) is a polyphenol-rich Brazilian fruit known to improve glucose metabolism. This study investigated its effects on brown (BAT), inguinal (iWAT), and epididymal (eWAT) adipose tissues in a diet-induced obesity model. Rats received control (CT/TUC-), high-fat (HF/TUC-), control with Tucum-do-Cerrado (CT/TUC+), or high-fat with Tucum-do-Cerrado (HF/TUC+) diets. Tucum-do-Cerrado promoted healthier adipose expansion by stimulating adipogenic/thermogenic-related genes while downregulating lipogenesis. In BAT, HF/TUC+ attenuated adipocyte hypertrophy and decreased the number of unilocular adipocytes. Tucum-do-Cerrado downregulated Fasn mRNA levels and glutathione peroxidase (GPx) activity, whereas it increased thermogenesis-related Ucp1 mRNA. In the iWAT, HF/TUC+ decreased the adipocyte area and oxidative markers, while increasing the number of multilocular adipocytes and UCP1 protein levels. Tucum-do-Cerrado decreased Acaca, Prkaa2, Il10 mRNA, and GPx activity; conversely, it stimulated the thermogenesis-related genes Ppargc1a and Prdm16. Regarding eWAT, its consumption increased the number of multilocular adipocytes, Prkaa1, Ppargc1a, Ucp1, Prdm16, and Vegfa mRNA levels, carbonyl levels, and GPx activity, while decreasing Acaca and Fasn mRNA levels. Therefore, Tucum-do-Cerrado prevented adipocyte hypertrophy and increased the number of multilocular adipocytes. The beneficial effects of Tucum-do-Cerrado in an obesity model may be attributed to enhanced antioxidant capacity and inflammatory status, highlighting its potential in attenuating adipose tissue dysfunction.

To investigate the effect of Wenyang Huazhuo Tongluo Formula (WHT) for promoting angiogenesis in systemic sclerosis (SSc) dermal microvascular endothelial cells and its possible mechanism. Wistar rats were gavaged with 15, 30, or 60 g/kg WHT and normal saline for 7 consecutive days to prepare low-, medium-, or high-concentration WHT-medicated sera and blank serum, respectively. Human dermal microvascular endothelial cells (HDMECs) in routine culture were treated with the sera from SSc patients to establish an SSc cell model. The cells were treated with the blank serum, low-, medium-, or high-concentration WHT-medicated sera, or blank serum combined with EGCG (a Sema3A inhibitor). The changes in cell proliferation, migration and angiogenesis were assessed using CCK-8 assay, wound-healing assay and Matrigel tube formation assay, and the mRNA and protein expressions of Sema3A, Nrp1, VEGFA, CD31 and α-SMA were analyzed using qRT-PCR and Western blotting. HDMECs treated with the serum from SSc patients exhibited significantly reduced cell proliferation and migration rates, increased α-SMA, Sema3A and VEGFA protein and mRNA expression levels, decreased CD31 and Nrp1 protein and mRNA expression levels, and suppressed angiogenesis. In SSc serum-induced cells, treatments with low-, medium-, or high-concentration WHT-medicated sera or EGCG all significantly improved cell survival and migration rates, reduced α-SMA, Sema3A and VEGFA protein and mRNA expression levels, enhanced CD31 and Nrp1 protein and mRNA expressions, and promoted angiogenesis of the cells. WHT promotes angiogenesis of SSc sera-induced HDMECs by modulating the Sema3A/Nrp1 signaling pathway.

Spexin as a modulator of luteal homeostasis: in vitro effect on the regulation of markers of angiogenesis, cell survival and autophagy pathways in porcine corpus luteum.

Nerve guidance conduits (NGCs) hold considerable value in the field of nerve regeneration, yet current approaches display constraints regarding the diversity of loaded factors and their short sustained-release duration. Herein, a type of multifunctional inverse opal NGC integrating bone marrow mesenchymal stem cells (BMSCs) is presented for peripheral nerve repair. The generation of inverse opal scaffolds leverages the inversion of a monodisperse emulsion droplet template crafted by microfluidic technique. Ascribed to the biocompatibility and the cell-concentrating properties of the inverse opal scaffold, coupled with the inherent capabilities of BMSCs, such an NGC enables the secretion of nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor, promoting the migration of rat Schwann cells and differentiation of pheochromocytoma 12 cells. Reverse Transcription Quantitative Real-Time Polymerase Chain Reaction (RT-qPCR) further revealed that three-dimensional (3D) BMSC spheroid culture in the inverse opal scaffold significantly upregulated bFGF, PDGF, and VEGF mRNA expression relative to two-dimensional (2D) culture. Further in vivo experimentation confirms the promising efficacy of NGCs loaded with BMSCs in repairing 10 mm sciatic nerve defects. These results underscore the substantial potential of this innovative design for peripheral nerve regeneration.

The high-carbohydrate diet delayed skin wound healing through suppressing AGEs-MTOR-HIF1A-driven glycolysis in turbot, Scophthalmus maximus L.

A mechanistic computational model of the HIF signaling pathway in endothelial cells

Gastric cancer (GC) remains one of the most common malignant tumors worldwide, with high incidence and mortality rates. Angiopoietin-like protein 1 (ANGPTL1), a member of the ANGPTL family, is known to function as an anti-angiogenic factor and tumor suppressor. However, its role and underlying mechanisms in GC development have not been investigated and require further investigation. Protein expression levels were analyzed using western blotting and immunofluorescence (IF) assays. Cell viability was assessed using the CCK-8 assay, and cell proliferation was evaluated through colony formation assays. Cell migration and invasion were examined using Transwell assays. Angiogenic capacity was determined through tube formation assays. The VEGFA mRNA expression was detected through RT-qPCR. The level of VEGFA was confirmed through ELISA. The tumor size, volume and weight were confirmed through the in vivo assay. The CD31 protein expression was verified though IHC assay. ANGPTL1 was found to be expressed at lower levels in GC cells, and negatively correlated with VEGFA. ANGPTL1 significantly inhibited GC cell proliferation, migration, and invasion. Furthermore, ANGPTL1 suppressed angiogenesis in vitro. Mechanistically, it was observed that ANGPTL1 overexpression reduced VEGFA expression, and ANGPTL1 can interact with VEGFA. Importantly, reintroduction of VEGFA reversed the inhibitory effects of ANGPTL1 on GC progression. Lastly, VEGFA overexpression retarded the tumor growth in vivo. This study demonstrates that ANGPTL1 inhibits the growth, migration, and angiogenesis of GC cells by downregulating VEGFA expression. These findings suggest that ANGPTL1 may serve as a promising therapeutic target for gastric cancer treatment.

Dynamic linking bone ECM-mimic hydrogel for anti-inflammatory therapy of cranial defect.

Thrombocytopenia is a common complication of various clinical conditions, resulting from impaired megakaryocyte function, reduced platelet production, or excessive platelet destruction. Current treatments, including platelet transfusions and thrombopoietin receptor agonists, are limited by platelet supply constraints and risks such as thrombotic complications. Emerging research highlights the role of mitochondrial-related biological processes or components in thrombopoiesis, yet targeted therapeutics remain scarce. In this study, we investigated mitochondria-targeted compounds for their potential to enhance megakaryocyte and platelet productionusing a fetal liver megakaryocyte differentiation and platelet culture system, and explored their effects on in vivo megakaryocyte and platelet production using a radiation damage-induced thrombocytopenia mouse model. Our findings identify PTC299, a dihydroorotate dehydrogenase and VEGFA mRNA translation inhibitor, as a promising pro-plateletogenic agent. PTC299 not only enhances megakaryocyte and platelet production in vitro but also accelerates their recovery in a mouse model of radiation-induced thrombocytopenia. Additionally, PTC299 alleviates irradiation-induced splenomegaly. PTC299 promotes megakaryocyte differentiation and platelet generation both in vitro and in vivo, demonstrating potential values for thrombocytopenia treatment and platelet regeneration.

Ulcerative colitis is a chronic inflammatory bowel disease characterized by inflammation of the colonic mucosa. This study aimed to evaluate the effects of dexpanthenol in an acetic acid-induced model of colitis in rats. Thirty-two adult male Wistar rats were randomly assigned to four groups: Control (intracolonic saline), dexpanthenol, acetic acid (A.A; 4% acetic acid-induced colitis), and treatment (A.A + dexpanthenol 500). Colitis was induced by intracolonic administration of 4% acetic acid. After 7 days, macroscopic and histopathological evaluations of the colon were performed. The mRNA expression levels of IL-6 and VEGF were assessed using real-time PCR. Stool consistency, colon length, and colon weight-to-length ratio were also measured. Dexpanthenol administration significantly ameliorated acetic acid-induced colitis. The A.A + dexpanthenol group showed partial restoration of colon length, improved stool consistency, and a reduced colon weight-to-length ratio compared with the A.A group. Histopathological analysis demonstrated reduced mucosal damage and decreased inflammatory cell infiltration in dexpanthenol-treated rats. In addition, dexpanthenol significantly decreased IL-6 and VEGF mRNA expression compared with the A.A group (P < 0.0001). Dexpanthenol exerts protective effects against experimental ulcerative colitis in rats by attenuating inflammation, improving tissue integrity, and downregulating IL-6 and VEGF gene expression. These findings suggest its potential as a supportive therapeutic agent in ulcerative colitis.

Spine fusion devices fabricated from polyether ether ketone (PEEK) using traditional machining or with surface pores created by salt leaching result in a fibrous connective tissue interface. To overcome this limitation, we used fused strand deposition (FSD), which elevates fused filament fabrication (FFF) or fused deposition modeling (FDM) to generate a first-of-its-kind PEEK implant with an architecture designed to mimic trabecular bone. We examined the responses of human bone marrow stromal cells (MSCs) and macrophages isolated from the femurs of C57/Bl6 male mice to the additively manufactured porous PEEK (PP) and porous PEEK coated with HA (PP-HA) to determine if these modifications would improve cell response compared to solid PEEK (SP). PP and PP-HA constructs had similar 3D architectures but differed in hydrophilicity (PP-HA>PP). MSCs and macrophages were cultured on PP, PP-HA, and SP, and osteoblast differentiation and M1/M2 polarization were assessed. MSCs attached to the SP surface and the PP and PP-HA fibers, and synthesized osteoblast proteins in a surface-dependent manner. Notably, MSCs and macrophages produced VEGF mRNA and protein on PP-HA at levels higher than those on PP or SP. Macrophages grown on PP and PP-HA exhibited reduced expression of pro-inflammatory cytokines compared to cells on SP and increased levels of anti-inflammatory cytokines, but they did not exhibit a distinct M1 or M2 phenotype. These results show that additive manufacturing of a unique, fully porous PEEK implant using FSD, results in a surface that promotes MSC differentiation and decreases the pro-inflammatory response of macrophages to the surface, suggesting that PP and PP-HA implants will improve regeneration and eventually osseointegration in vivo. Importantly, the mechanisms involved are not the same as would be expected when testing Ti6Al4V substrates under the same experimental conditions and may have been obscured if the cells had been cultured using osteogenic media (OM). We correlated our in vitro findings with the clinical use of PP-HA implants in four patients, each of whom was treated with a different bone graft material; in all cases, fusion was achieved.

Modifying VEGF-A mRNA by combinatorial optimization to enhance therapeutic efficacy for myocardial infarction

Background: Herpes simplex virus 1 (HSV-1) establishes lifelong latency with periodic reactivation, and is of particular concern in immunocompromised patients, such as those receiving treatment with chemotherapy. Experimental data suggest that upon infection, HSV-1 induces the expression of vascular endothelial growth factor A (VEGF-A), a key mediator of angiogenesis and inflammation. However, the link between HSV-1 reactivation and physiological or systemic VEGF-A in patients receiving oncological therapy have not been examined broadly. Methods: A cross-sectional molecular study to investigate the relationship between HSV-1 reactivation and VEGF-A expression in Iraqi chemotherapy patients. Methods: 300 subjects (200 chemotherapeutic patients and 100 control not receiving chemotherapy) were recruited. HSV-1 status was assessed by serology (IgG, IgM) and qPCR detection of viral DNA in oral swabs and plasma. Enzyme-linked immunosorbent assay (ELISA) was used to measure serum VEGF-A concentrations and reverse transcription qPCR was performed to quantify peripheral blood mononuclear cells levels of VEGFA mRNA. A multivariable linear regression was conducted to determine the independent relationship between positivity for HSV-1 DNA and log-transformed VEGF-A levels while controlling for demographic, clinical, and immunological variables. Results: HSV-1 DNA was found in 18% of patients undergoing chemotherapy versus 5% of controls (p &lt; 0.001). Serum VEGF-A levels in chemotherapy patients were significantly higher than controls (median 412 according to 221 pg/mL; p &lt; 0.001). In chemotherapy patients, individuals who were positive for HSV-1 DNA had significantly higher serum VEGF-A (median 712 with 368 pg/mL, p &lt; 0.001) and increased expression of VEGFA mRNA (median fold change 3.94 compare with 1.63, p &lt; 0.001). Oral viral load positively correlated with serum VEGF-A (Spearman ρ = 0.58, p &lt; 0.001). In a multivariable analysis, positivity for HSV-1 DNA remained independently associated with higher log (VEGF-A) (β = 0.41, 95% CI 0.28–0.54, p &lt; 0.001). Conclusions: Reactivation of HSV-1 is independently correlated with enhanced VEGF-A expression in un-females and non-HSV infected chemotherapy doomed fatalities. This observation hinted that viral reactivation upon immune suppression might drive systemic angiogenic and inflammatory signaling. The clinical implications of HSV-1–mediated VEGF-A upregulation in oncology settings warrant examination in prospective studies. In the state of virus reactivation, qPCR was used to quantify HSV-1 and VEGF-A expression level. Keywords: HSV-1, VEGF-A, chemotherapy, angiogenesis, qPCR, Iraq, viral reactivation

Effective management of corneal neovascularization (CoNV) remains challenging, and the role of epitranscriptomic regulation, particularly N5-methylcytosine (m5C) modification, in this process remain incompletely defined. This study investigated the function and mechanism of the RNA-binding protein YBX1 in CoNV following alkali burn (AB). An AB-induced CoNV model was generated using C57BL/6 mice. In vitro, human umbilical vein endothelial cells (HUVECs) underwent hypoxia/reoxygenation (H/R). Multi-omics approaches including transcriptome sequencing, RNA immunoprecipitation sequencing, and m5C methylated RNA immunoprecipitation sequencing were used to identify YBX1 targets and their modification status. Functional assays assessed angiogenesis, apoptosis, and reactive oxygen species (ROS). The therapeutic potential of the YBX1 inhibitor Soyasaponin II (SII) was evaluated in vivo. YBX1 was upregulated following AB and H/R. YBX1 knockdown suppressed HUVEC migration, tube formation, and ROS production, while promoting apoptosis; these effects were rescued by HIF-1α overexpression. Mechanistically, YBX1 activated the JAK1/STAT3 pathway and recognizes m5C-modified sequences on STAT3 and VEGFA mRNAs, enhancing their stability. In vivo, subconjunctival injection of SII attenuated CoNV, reduced inflammation, and modulated macrophage polarization. Our study unveils a novel epitranscriptomic mechanism in which YBX1 drives CoNV by regulating the stability of m5C-modified STAT3 and VEGFA mRNAs, thereby activating the JAK1/STAT3/HIF-1α axis. Inhibition of YBX1 with SII effectively counteracts this pathway, highlighting YBX1 as an attractive candidate for intervention against sight-threatening CoNV.

This study aimed to evaluate FOXC1-mediated regulatory mechanisms on gene and protein expression profiles in primary human limbal epithelial cells (pLECs) using siRNA-mediated FOXC1 knockdown under basal conditions and following lipopolysaccharide (LPS) and interleukin-1β (IL-1β)-induced inflammatory conditions. The gene expression related to inflammation, epithelial differentiation, cell proliferation and remodeling, and retinoic acid metabolism was analyzed using qPCR. Corresponding protein levels were assessed through Western blotting and ELISA. FOXC1 silencing significantly downregulated epithelial differentiation markers KRT12 and KRT13 at the mRNA and protein levels (p ≤ 0.045), whereas KRT3 and KRT19 were unaffected. Inflammatory signaling was markedly altered, with a reduced IL-6 and IL-8 mRNA expression (p ≤ 0.029), increased IL-1α expression (p ≤ 0.015), and condition-dependent changes in IL-6 and IL-8 protein secretion. CCL2 was increased at the mRNA level only (p = 0.007). VEGFA mRNA was consistently reduced (p ≤ 0.022) without corresponding protein changes, while TGF-β protein was increased under non-inflammatory and LPS conditions (p ≤ 0.011). Genes involved in retinoid metabolism, including CYP1B1, FABP5, CRABP2, RDH10, STRA6, and ALDH3A1, were significantly downregulated (p ≤ 0.037), with reduced CRABP2 and RDH10 protein levels (p ≤ 0.017) and a decreased FABP5/CRABP2 ratio under IL-1β stimulation (p = 0.006). FOXC1 knockdown affected proliferation-related genes, with decreased FOSL2 (p = 0.048) and increased MKi67 (p = 0.006). FOXC1 silencing disrupts epithelial differentiation, inflammatory signaling, retinoid metabolism, and selected proliferation-related pathways at the transcriptional level, with more selective effects on protein levels. Such changes may potentially predispose the ocular surface to lineage instability, fibrosis, and impaired regenerative capacity.

Glioblastoma (GB) is the most aggressive primary glioma, with a median survival of 15-18 months. Current treatments are often ineffective, largely due to tumor heterogeneity and recurrence. Advances in understanding GB's molecular landscape and microenvironment have highlighted new therapeutic strategies to fight this life-threatening tumor. Given the pivotal role of natural compounds in drug discovery, those with anti-inflammatory and cytotoxic/cytostatic properties are emerging as promising candidates for GB therapy. This study investigates the antitumor and immunomodulatory effects of 7α-acetoxy-6β-hydroxyroyleanone (Roy), a diterpene isolated by our team from Plectranthus hadiensis Schweinf., using both 2D and 3D GB cell models. U87 cells were used as a standard GB model and to generate monocellular and multicellular spheroids (U87, HMC3, and/or HBMEC cells). Both models were treated with 16 µM of Roy, a concentration previously shown to be tumor-specific. Roy significantly reduced spheroid size and metabolic activity over time, with the most pronounced effects observed in multicellular spheroids. This compound also inhibited cell proliferation by preventing colony formation and downregulating CDK4 and VEGFA mRNA levels. Roy's bioactivity was enhanced in the presence of conditioned medium (secretome from GB and/or microglia cells), exerting a neuromodulatory effect by modulating IL6/JAK2/STAT3 mRNA expression and by suppressing the secretion of cytokines involved in the chronic inflammatory state within the GB microenvironment. Importantly, Roy was also able to cross the blood-brain barrier. These findings, in line with our previous work, underscore the cytotoxic potential of this natural compound, suggesting Roy as a promising lead candidate for future GB treatment strategies.

Ferroptosis plays a crucial role in lung diseases, including bronchopulmonary dysplasia (BPD), but its involvement in BPD pathogenesis and impact on pulmonary vascular development remain unclear. This study investigated ferroptosis in BPD and evaluated sodium propionate’s (SP) therapeutic potential in regulating ferroptosis and promoting pulmonary angiogenesis. Serum markers of ferroptosis and oxidative stress were measured in preterm infants (< 32 weeks) with and without BPD. BPD rats and human umbilical vein endothelial cell (HUVEC) models were established through hyperoxia induction. Sprague-Dawley rats were randomly assigned to four experimental groups: normoxia control (CON), normoxia with sodium propionate treatment (CON + SP), hyperoxia-induced BPD model (BPD), and hyperoxia with SP treatment (BPD + SP). In vitro experiments, HUVEC cells were respectively treated with hyperoxia and SP, and transfected with siRNA to detect the role of the SLC7A11/GPX4 pathway. Ferroptosis was evaluated by measuring lipid peroxidation (LPO), malondialdehyde (MDA), reactive oxygen species (ROS), and glutathione (GSH). Angiogenesis-related genes (VEGFA, CD31) and angiogenic ability were evaluated by Western blot, RT-PCR and in vitro experiments. Serum LPO and iron levels were significantly elevated in BPD infants. SP alleviated lung injury in BPD rats. SP significantly inhibited PTGS2 expression, decreased ROS, MDA, and LPO levels, and increased GSH in BPD models. It also enhanced mRNA and protein expression of SLC7A11, GPX4, VEGFA, and CD31. Silencing SLC7A11 or GPX4 counteracted SP’s protective effects in hyperoxia-exposed HUVECs. Ferroptosis contributes to BPD pathogenesis. SP effectively inhibits ferroptosis and promotes angiogenesis in experimental BPD models, suggesting a novel avenue for potential combination therapies in BPD treatment.

GABA receptors are classically known for driving neuronal hyperpolarization and modulating synaptic transmission. In glial cells, however, GABA induces depolarization and triggers calcium-dependent signaling pathways. Müller glia, the principal retinal glial population, maintain retinal homeostasis and are the major source of neuroretinal VEGF-A, a key angiogenic factor in development and disease. Although GABA receptor (GABAR) activity has been proposed to influence retinal VEGF-A, it remains unclear whether this regulation occurs through Müller glial cells (MGC) and which mechanisms are involved. Here, we investigated how GABAR activation modulates VEGF-A in primary mouse MGC cultures. Cells were exposed to GABA and selective agonists or antagonists of GABAA (muscimol, gabazine) and GABAB receptors (baclofen, CGP55845). VEGF-A expression and secretion were analyzed by immunofluorescence, western blot, RT-qPCR, and ELISA. To assess Ca2+ involvement, we used Ca2+-free Ringer-Krebs solution and the L-type channel blocker nimodipine, and examined MAPK signaling with the ERK1/2 inhibitor FR180204. Our findings show that GABA and muscimol increased VEGF-A fluorescence intensity after 48 hours while reducing VEGF-A secretion, without altering Vegfa mRNA. Both effects were abolished by extracellular Ca2+ removal or nimodipine, indicating a Ca2+-dependent mechanism. FR180204 also attenuated GABA- and GABAA-mediated effects, implicating MAPK signaling. Short-term assays revealed that GABA rapidly elevates VEGF-A protein and secretion within ∼30 minutes. Together, these findings identify a Ca2+- and GABAA-dependent pathway through which Müller glia regulate VEGF-A production and release, providing new insight into glial signaling and neurotransmitter-driven modulation of retinal angiogenic factors.

Background: recently, regenerative medicine has introduced a new branch of science that facilitates the repair of damaged tissues and organs in acute myocardial infarction. This study explores the role of the C-type natriuretic peptide (CNP) system in myocardial infarction (MI) and its modulation by Apelin-13 functionalized patches (A-13p). Methods: using an experimental rat model of ischemia/reperfusion, the rats were divided into four groups: Sham, Infarct, Sham with A-13p, and Infarct with A-13p. Cardiac tissue from the infarct, border, and remote zones was analyzed for CNP and its receptors' mRNA expression via Real-Time PCR. Results: histological analysis, 4 weeks post A-13p implantation, showed no damage from A-13p implantation in either MI or Sham groups, with reduced left ventricle wall thinning in the Infarct group treated with A-13p. CNP mRNA expression was higher in the infarcted groups (p = ns), especially in the border/infarct zone (BZ + IZ), compared to the Sham group (p = 0.05). NPR-B receptor expression was higher in the RZ than in (BZ + IZ), both in the absence (p = 0.02) and presence of patches (p = 0.01), while NPR-C expression was lower. No significant differences were observed in VEGF mRNA levels across the groups. Conclusions: the findings suggest that the CNP system is involved in MI and that A-13p modulates CNP expression, highlighting CNP as a potential target for therapeutic strategies aimed at regulating vascular remodeling and angiogenesis in MI treatment.

IntroductionRetinopathy of prematurity (ROP) is a vasoproliferative retinal disorder in preterm infants driven by impaired vascular development, hypoxia, oxidative stress, and pathological angiogenesis. Current treatments mainly target VEGF signaling but do not directly address upstream oxidative injury. Hepcidin, a key iron-homeostasis regulator, may modulate redox balance and angiogenic activation. However, whether hepcidin is reduced in ROP and whether supplementation mitigates hypoxia-induced oxidative stress and abnormal angiogenesis in retinal endothelial cells remain unclear.ObjectiveTo investigate the effects of hepcidin on human retinal microvascular endothelial cells (hRMECs) under oxidative stress and assess its translational potential in ROP.MethodsClinical data were collected from 35 preterm infants with gestational age <36 weeks, including 24 non-ROP controls and 11 ROP infants. Gestational age, birth weight, ROP stage, and serum hepcidin were analyzed. hRMECs were exposed to 1% O2 to establish a hypoxia model. Cell viability, VEGFA and HIF-1α mRNA expression, secreted VEGFA, intracellular reactive oxygen species (ROS), transcriptomic changes, and tube formation were evaluated. Hepcidin was added under hypoxia to assess its effects on oxidative stress and angiogenic activation.ResultsInfants with ROP had lower gestational age and birth weight than controls, and gestational age differed across ROP stage groups. Serum hepcidin-related signals were numerically lower in ROP infants, but no significant stage-dependent decrease was observed. In hRMECs, 1% O2 altered cell viability, upregulated VEGFA and HIF-1α mRNA, increased secreted VEGFA, and promoted ROS accumulation, confirming a hypoxia-induced endothelial stress model. Transcriptomic analysis suggested that hepcidin affected hypoxia-responsive metabolic pathways, including cysteine and methionine, selenocompound, and tryptophan metabolism. Functional validation showed that hepcidin reduced VEGFA at transcript and secreted protein levels, decreased intracellular ROS, and suppressed hypoxia-induced tube formation by reducing junction points, branch points, and total tube length.ConclusionThis study provides preliminary clinical and experimental evidence supporting a potential role of hepcidin-related signaling in ROP. Although the clinical findings are exploratory, the in vitro results indicate that hepcidin can attenuate hypoxia-induced oxidative stress and pathological angiogenic activation in retinal endothelial cells. Hepcidin may represent a biologically relevant pathway and potential candidate for further biomarker and therapeutic research in ROP.