Human Umbilical Vein Endothelial Cells Research Articles

Differential Mechanisms of Soybean-Derived ACE2-Activating Peptides IVPQ and IAVPT in ACE2-Mediated Endothelial Protection.

This study aimed to investigate the modulatory effects of soybean-derived peptides IVPQ and IAVPT, which were initially identified as potent ACE2-activating peptides, on Ang II-induced endothelial dysfunction in human umbilical vein endothelial cells (HUVECs) and the underlying mechanisms via ACE2 activation. IVPQ and IAVPT ameliorated Ang II-induced malignant migration and NO reduction in HUVECs via the activation of the ACE2/Ang-(1-7)/MasR axis, resulting in Ang II degradation and decreased Ang II signaling. These protective effects were attenuated by ACE2 knockdown to different degrees, which was possibly due to different mechanisms of activating ACE2, where IAVPT directly activated ACE2 at a concentration of 1.0 × 10-4 M and IVPQ upregulated ACE2 likely through effects on ACE2 mRNA stability. These results contributed to our understanding of the mechanism of ACE2-activating peptides regulating endothelial function.

MT2A promotes angiogenesis in chronically ischemic brains through a copper–mitochondria regulatory mechanism

BackgroundApproximately half of patients with chronic ischemic cerebrovascular disease (CICD) exhibit poor revascularization. Metallothionein 2 A (MT2A) has a high affinity for metal ions and is potentially capable of chelating toxic copper ions to alleviate the impairment of angiogenesis. Therefore, we hypothesized that MT2A could promote angiogenesis in chronically ischemic brains by neutralizing excessive copper ions during copper overload (CPO).MethodsWe first collected dura matter (DM) samples from CICD patients and examined the expression of cuproptosis-related genes (DLAT, FDX1, and SDHB) to confirm the inhibitory effect of CPO on angiogenesis. Then, we treated human umbilical vein endothelial cells (HUVECs) with different concentrations of elesclomol and CuCl2 to determine the optimal concentration for inducing CPO. HUVEC activity and mitochondrial structure and function were detected to explore the ability of MT2A to alleviate CPO-induced damage. Finally, a rat model of 2-vessel occlusion plus encephalo-myo-synangiosis (2VO + EMS) with CPO was established to test the proangiogenic effect of MT2A through the copper–mitochondria regulatory mechanism in chronically ischemic brains.ResultsCompared with those from Matsushima grade A patients, DM samples from Matsushima grade C patients presented significantly greater DLAT and FDX1 expression and significantly lower SDHB expression. The optimal drug concentration for inducing CPO was subsequently determined, and in vitro experiments revealed that HUVEC activity was significantly decreased in the CPO group under hypoxic culture, accompanied by increased DLAT oligomerization, decreased SDHB expression, increased HSP70 expression. Moreover, significantly more common mitochondrial aberrations and significantly lower mitochondrial activity were detected in the CPO group compare with the control group. Additionally, MT2A overexpression alleviated CPO-induced mitochondrial dysfunction and cytotoxicity, improving HUVEC viability. In vivo, a CPO rat model was established, and CPO inhibited cerebral angiogenesis in 2VO + EMS model rats. Moreover, significantly greater CD31 expression, less DLAT accumulation, more mitochondria, and fewer mitochondrial abnormalities were observed in the CPOMT2A+ group than in the CPO group, accompanied by significantly improved cerebral blood perfusion and cognitive function.ConclusionMT2A can promote angiogenesis in chronically ischemic brains by neutralizing excessive copper ions and rescuing CPO-induced mitochondrial dysfunction.

An Endogenous Aryl Hydrocarbon Receptor Ligand Dysregulates Endothelial Functions, Transcriptome, and Phosphoproteome.

We have reported that an endogenous aryl hydrocarbon receptor (AhR) ligand, 2-(1'H-indole-3'-carbonyl)-thiazole-4-carboxylic acid methyl ester (ITE), inhibits functions of human umbilical vein endothelial cells (HUVECs) and induces preeclampsia (PE)-like symptoms in rats. Herein, we tested the hypothesis that ITE impairs endothelial functions via disturbing transcriptome and phosphoproteome in HUVECs. We measured AhR activity in human maternal and umbilical vein sera from PE and normotensive (NT) pregnancies. The serum-induced changes in CYP1A1/B1 mRNA (indexes of AhR activation) in HUVECs were quantified using RT-qPCR. ITE's effects on endothelial proliferation and monolayer integrity in female and male HUVECs were determined. We profiled ITE-induced changes in transcriptome and phosphoproteome in HUVECs using RNA-seq and bottom-up phosphoproteomics, respectively. After 12 hr of treatment, umbilical vein sera from PE increased CYP1A1 mRNA (1.7-fold of NT) HUVECs, which was blocked by CH223191, an AhR antagonist. ITE dose-dependently inhibited endothelial proliferation (76%-87% of control) and time-dependently reduced endothelial integrity with a maximum inhibition (~ 10%) at 40 hr. ITE induced 140 and 80 differentially expressed genes in female and male HUVECs, respectively. ITE altered phosphorylation of 92 and 105 proteins at 4 and 24 hr, respectively, in HUVECs. These ITE-dysregulated genes and phosphoproteins were enriched in biological functions and pathways are relevant to heart, liver, and kidney diseases, vascular functions, and inflammatory responses. Thus, endogenous AhR ligands may impair endothelial functions by disturbing transcriptome and phosphoproteome. These AhR ligand-dysregulated genes and phosphoproteins may be therapeutic and cell sex-specific targets for PE-induced endothelial dysfunction.

PCSK9 affects vascular senescence through the SIRT1 pathway.

Age is an independent risk factor for atherosclerotic cardiovascular disease that increases the susceptibility of older adults to vascular intimal thickening, endothelial dysfunction, and thrombosis. However, the mechanism underlying vascular injury is not fully understood. In the present study, the effect of proprotein convertase subtilin-type kexin 9 (PCSK9) inhibitors on the senescent state of human umbilical vein endothelial cells (HUVECs) and on senescent mice and lipopolysaccharides (LPS) were assessed. The senescent state of mice was delayed under PCSK9 inhibitor treatment, and the expression of P16, P21, and P53 proteins in senescent cells was increased because LPS induction stimulated PCSK9 activation. PCSK9 overexpression accelerated cell senescence, activated a large number of oxidative stress pathways, and increased the expression of senescence-related genes (including P16, P21, and P53). In addition, inhibition of the sirtuin 1 (SIRT)1 oxidative stress pathway can attenuate the aging-promoting effects of PCSK9, which are elevated as a result of LPS induction. The SIRT1 activator was more efficient than LPS alone in inducing the expression of senescence-related genes. Therefore, PCSK9 inhibitors can delay the aging of the vascular by reducing cellular SIRT1 levels. Therefore, it can be concluded that PCSK9 inhibition inhibits vascular senescence by reducing the expression of senescent proteins by regulating the SIRT1 pathway.

Evaluation of the Potential In Vitro effects of Plantago major L. on Wound Healing in Human Umbilical Vein Endothelial Cells (HUVEC)

The treatment of skin wounds remains a major concern in the field of medicine, particularly in the case of chronic wounds resulting from various disorders such as diabetes. The utilization of herbs or herbal preparations for the purpose of healing skin wounds presents a therapeutic challenge within the realm of traditional medicine. Plantago major L. is known to have bioactive compounds that have wound healing activity such as aucubin. This study aimed to determine the in vitro wound healing potential of Plantago major L. extract (PLE). The study involved several assays, including phytochemical examination of PLE using TLC, cell viability testing using MTT assay, and wound healing testing using scratch assay on human umbilical vein endothelial cells (HUVEC). The results confirmed the presence of aucubin as one of the compounds in PLE. It was observed that PLE with 125 μg/mL exhibited the highest wound closure percentage at 90.66%. This study shows that PLE possesses wound healing capabilities.Keywords: Plantago major L., PLE, cytotoxic assay, wound scratch assay, HUVEC.

Metformin exerted tumoricidal effects on colon cancer tumoroids via the regulation of autophagy pathway

BackgroundDespite the existence of promising outcomes from standard 2D culture systems, these data are not completely akin to in vivo tumor parenchyma. Therefore, the development and fabrication of various 3D culture systems can in part mimic intricate cell-to-cell interaction within the real tumor mass. Here, we aimed to evaluate the tumoricidal impacts of metformin (MTF) on colorectal cancer (CRC) tumoroids in an in vitro system via the modulation of autophagy.MethodsCRC tumoroids were developed using human umbilical vein endothelial cells (HUVECs), adenocarcinoma HT29 cells, and fibroblasts (HFFF2) in a ratio of 1: 2: 1 and 2.5% methylcellulose. Tumoroids were exposed to different concentrations of MTF, ranging from 20 to 1000 mM, for 72 h. The survival rate was detected using an LDH release assay. The expression and protein levels of autophagy-related factors were measured using PCR array and western blotting, respectively. Using H & E, and immunofluorescence staining (Ki-67), the integrity and proliferation rate of CRC tumoroids were examined.ResultsThe current protocol yielded typical compact tumoroids with a dark central region. Despite slight changes in released LDH contents, no statistically significant differences were achieved in terms of cell toxicity in MTF-exposed groups compared to the control tumoroids, indicating the insufficiency of MTF in the induction of tumor cell death (p > 0.05). Western blotting indicated that the LC3II/I ratio was reduced in tumoroids exposed to 120 mM MTF (p < 0.05). These data coincided with the reduction of intracellular p62 content in MTF 120 mM-treated tumoroids compared to MTF 40 mM and control groups (p < 0.05). PCR array analysis confirmed the up-regulation, and down-regulation of several genes related to various signaling transduction pathways associated with autophagy machinery and shared effectors between autophagy and apoptosis in 40 and 120 mM MTF groups compared to the non-treated control group (p < 0.05). These changes were more prominent in tumoroids incubated with 120 mM MTF. Histological examination confirmed the loosening integrity of tumoroids in MTF-treated groups, especially 120 mM MTF, with the increase in cell death via the induction of apoptosis (chromatin marginalization) and necrotic (pyknotic nuclei) changes. In the 120 mM MTF group, spindle-shaped cells with the remnants of a fibrillar matrix were detected. Data indicated the reduction of proliferating Ki-67+ cells within the tumoroids by increasing the MTF concentration from 40 to 120 mM.ConclusionsDifferent shared autophagy/apoptosis genes were modulated in CRC tumoroids after MTF treatment coinciding with both typical necrotic and apoptotic cells within the tumoroid structure. MTF can inhibit the integrity and proliferation of CRC tumoroids in dose-dependent manner.

Exosomes Derived from Dialysis Patients Serum enhance Endothelial-Mesenchymal Transition and Calcification in Endothelial Cells.

Vascular calcification (VC) is prevalent among patients with end stage kidney disease (ESKD). Exosomes, small extracellular vesicles actively secreted by cells, contain proteins, nucleic acids, lipids and other bioactive substances and are considered major mediators of cell-cell interactions. Endothelial-Mesenchymal Transition (EndMT) has been observed in a variety of pathological conditions, such as abnormal shear stress, vascular damage, and chronic inflammation. The aim of this research was to assess the effects of serum-derived exosomes from ESKD patients with VC on the induction of EndMT in endothelial cells and their potential role in accelerating VC. Twenty hemodialysis patients with VC and 10 healthy volunteers were recruited. Cardiac and brain VC were assessed among patients with ESKD treated with dialysis. Serum samples were taken at dialysis initiation for exosome isolation. Human umbilical vein endothelial cells (HUVEC) were treated with 100 µg/mL exosomes for 24-96 hours. At the end of incubation, cells were collected for mRNA and protein analysis. Exosomes isolated from dialysis patients with VC induced EndMT in HUVECs. After 24h, endothelial markers CD31 and VE-cadherin were decreased (31% and 51%, respectively; P<0.001) and the mesenchymal proteins Vimentin and N-cadherin were increased (283% and 156%, respectively; P<0.001), compared to healthy exosomes. After 96h of incubation, expression of genes essential for osteoblast differentiation, including the bone morphogenetic genes (BMP2, BMPR2, BMP4 and BMP9), and the transcription factor RUNX2 were significantly elevated. Exosomes derived from the serum of dialysis patients with VC, induced EndMT and contributed to calcification. The vicious cycle highlighted the intricate interplay between exosomes, EC and VC, emphasizing the critical necessity for therapeutic strategies to disrupt this pathway and mitigate calcification advancement.

PH-Responsive Polyglycerol Nanogels for Periodontitis Treatment through Antibacterial and Pro-Angiogenesis Action.

Periodontitis is a microbe-driven inflammatory disease leading to bone resorption and tissue destruction. We propose a dual-functional nanogel complex armed with the antimicrobial drug triclosan (TCS) and the pro-angiogenesis medication deferoxamine (DFO) for combating microbial pathogens and promoting tissue regeneration. The nanogel system (NG-TCS-DFO) that we fabricated from linear polyglycerol exhibits well-defined spherical morphology and a positively charged surface for bacteria adhesion. The rapid and sustained degradation of NG-TCS-DFO in the acidic environment of an infection site induces the on-demand release of TCS and DFO. The NG-TCS-DFO shows potent bacteria elimination of the gingivitis-causing bacteria Porphyromonas gingivalis in both planktonic (99.9 %) and biofilm (99 %) states. Furthermore, the NG-TCS-DFO can promote vascularization and migration of human umbilical vein endothelial cells (HUVECs). Contributing to the synergistic effect of TCS and DFO, the NG-TCS-DFO demonstrates significant bone tissue regeneration and accelerated healing of periodontitis in vivo. This polyglycerol-based nanogel may therefore offer smart combined delivery of multiple therapeutics against bacteria-driven diseases.

PH‐responsive Polyglycerol‐Nanogele zur Behandlung von Parodontitis durch antibakterielle und pro‐angiogenetische Wirkung

AbstractParodontitis ist eine entzündliche Erkrankung, die durch Mikroben ausgelöst wird und zu Knochenabbau sowie Gewebeschäden führt. In dem vorliegenden Artikel stellen wir einen Nanogel (NG)‐basierten Komplex vor, der zwei Funktion vereint: Er enthält den antimikrobiellen Wirkstoff Triclosan (TCS) und das pro‐angiogenetische Medikament Deferoxamin (DFO), um mikrobielle Erreger zu bekämpfen und die Geweberegeneration zu fördern. Das Nanogel (NG‐TCS‐DFO), das ausgehend von linearem Polyglycerin hergestellt wurde, weist eine wohldefinierte sphärische Morphologie auf und hat eine positiv geladene Oberfläche, die die Anhaftung von Bakterien begünstigt. Die schnelle und anhaltende Zersetzung des NG‐TCS‐DFO‐Komplexes in der sauren Umgebung eines Infektionsherdes bewirkt die gezielte Freisetzung von TCS und DFO. Das NG‐TCS‐DFO zeigt eine ausgeprägte Wirksamkeit bei der Eliminierung, der für Gingivitis verantwortlichen Bakterien Porphyromonas gingivalis, sowohl in planktonischer Form (99,9 %) als auch im Biofilm (99 %). Darüber hinaus fördert NG‐TCS‐DFO die Bildung von Blutgefäßen und die Migration von humanen Umbilikalvenen‐Endothelzellen (HUVECs). Durch die synergistische Wirkung von TCS und DFO zeigt der NG‐TCS‐DFO‐Komplex eine signifikante Regeneration des Knochengewebes und beschleunigt die Heilung von Parodontitis im Tiermodell. Dieses auf Polyglycerin basierende Nanogel bietet somit eine effiziente Kombinationstherapie zur Bekämpfung bakterienbedingter Erkrankungen.

The Beneficial Effects of CGK012 Against Lipopolysaccharide-Induced Inflammation.

This study investigates the protective effects of CGK012 [(7S)-(+)-cyclopentyl carbamic acid 8,8-dimethyl-2-oxo-6,7-dihydro-2H,8H-pyrano[3,2-g]chromen-7-yl-ester], a small-molecule inhibitor targeting the Wnt/β-catenin signaling pathway, against inflammatory responses elicited by lipopolysaccharide (LPS). The study evaluated the influence of CGK012 on heme oxygenase (HO)-1, cyclooxygenase (COX)-2, and inducible nitric oxide synthase (iNOS) expressions in LPS-stimulated human endothelial cells. It examined its effects on iNOS, tumor necrosis factor (TNF)-α, and interleukin (IL)-1β in LPS-challenged mice. CGK012 treatment resulted in increased HO-1 production, inhibited nuclear factor-kappa B activation, and decreased the levels of COX-2/PGE2 and iNOS/NO. Additionally, CGK012 reduced signal transducer and activator of transcription-1 phosphorylation and facilitated Nrf2 nuclear translocation and binding to antioxidant response elements, culminating in reduced IL-1β production in LPS-exposed human umbilical vein endothelial cells. Notably, the inhibitory effect of CGK012 on iNOS/NO was reversed upon HO-1 knockdown via RNA interference. In vivo, CGK012 markedly attenuated iNOS expression in lung tissue and decreased TNF-α levels in bronchoalveolar lavage fluid. These findings underscore the anti-inflammatory potential of CGK012, suggesting its therapeutic promise for conditions characterized by pathological inflammation.

Gelatin methacryloyl hydrogel encapsulating molybdenum-inspired macrophage-derived exosomes accelerates wound healing via immune regulation and angiogenesis.

Clinically, abnormal or delayed wound healing leads to functional disorders and disfiguring scars. A well-vascularized environment and an anti-inflammatory immune state are crucial during the healing process. Molybdenum (Mo) is an essential element for the human body in modulating metabolism, immune function and tissue repair. In this study, we found that Mo-inspired macrophages significantly promoted the proliferation, migration, tube formation, and the expression of angiogenic factors (VEGF, PDGF, CD31) in human umbilical vein endothelial cells (HUVECs). We then successfully isolated and characterized Mo-inspired macrophage-derived exosomes (Mo-Exo) and demonstrated their internalization by HUVECs, leading to enhanced proliferation, migration, tube formation, and angiogenic factor expression. Moreover, we proved that Mo-Exo could promote the polarization of macrophages from the pro-inflammatory M1 phenotype to the anti-inflammatory M2 phenotype. In terms of application, Mo-Exo encapsulated gelatin methacryloyl hydrogel (GelMA) exhibited notable potential in wound healing in rat skin defects, as evidenced by the reduced wound area and increased epidermal thickness observed in H&E and Masson's staining. Mechanically, Mo-Exo promoted angiogenesis through the activation of the ERK1/2 signaling pathway. In general, this research introduced a novel immunoregulatory and pro-angiogenic exosome hydrogel for clinical wound healing.

Dentin sialoprotein acts as an angiogenic factor through association with the membrane receptor endoglin.

Dentin sialophosphoprotein (DSPP) is highly expressed by odontoblasts, the cell type responsible for dentin formation. DSPP therefore has been extensively studied as a regulator of dentinogenesis. Besides defective dentinogenesis in teeth, Dspp deficient mice also display reduced blood vessels in the transition zone of femurs. However, the exact role and underlying mechanisms of DSPP in the process of blood vessel formation remain enigmatic. Here, we show that dentin sialoprotein (DSP), the NH2-terminal cleavage product of DSPP, promotes the migration and capillary-like structure formation of human umbilical vein endothelial cells (HUVECs) as well as the migration and endothelial differentiation of human dental pulp stem cells (DPSCs). Further experiments demonstrate that endoglin (ENG), a membrane receptor associated with angiogenesis, can be co-immunoprecipitated by DSP. Flow cytometry assays show that HUVECs and DPSCs, two cell types with endogenous ENG expression, display obvious binding signals of supplemented DSP protein, but human embryonic kidney 293T (HEK293T) cells, a cell type without endogenous ENG expression do not. Pretreatment with an anti-ENG antibody or knockdown of ENG inhibits the binding of DSP to DPSCs, while ENG overexpression enhances binding signals of DSP to HEK293T cells. Meanwhile, multiple experiments demonstrate that knockdown of ENG impairs DSP-induced migration and endothelial differentiation of DPSCs. Therefore, ENG is essential for the angiogenic effects of DSP. Moreover, Dspp deficient mice exhibit defective capillary formation in molars, supporting the positive role of DSP in blood vessel development. Collectively, these findings identify that DSP acts as an angiogenic factor through association with ENG.

Heparan sulfate acts in synergy with tight junction through STAT3 signaling to maintain the endothelial barrier and prevent lung injury development.

Damage to glycocalyx and tight junction are key determinants of endothelial permeability, which is the main pathological feature of acute respiratory distress syndrome (ARDS). However, the effect of glycocalyx heparan sulfate (HS) on tight junction proteins occludin and ZO-1 has not been revealed. In this study, the mice exposed to LPS results showed that FITC-albumin infiltration, HS shedding, and tight junction protein impairment were most severe at 6h of LPS treatment compared with those in other treatment times. The in vitro and vivo experiments revealed that tight junction damage, FITC-albumin infiltration, and pathological injury induced by LPS were significantly alleviated via protection of glycocalyx HS shedding. mRNA sequencing analysis demonstrated that the STAT signaling pathways played a crucial role in the inhibition of LPS-induced HS shedding in mice. Supplementation of exogenous HS in human umbilical vein endothelial cells (HUVECs) and mice ameliorated LPS-induced the tight junction barrier defect by inhibiting STAT3 phosphorylation. Further analysis uncovered that intervention of STAT3 signaling significantly alleviated LPS-induced tight junction proteins damage and vascular permeability in HUVECs and mice. Mechanistically, HS modulated tight junction proteins by STAT3 signaling, which might directly bind to the promoter regions of occludin and ZO-1. In conclusion, glycocalyx HS played an important role in protecting endothelial barrier function and preventing injury development, in synergy with tight junction through STAT3 signaling, which further alleviated pulmonary edema.

Assessment of silver-copper co-loaded mesoporous bioactive glass as an advanced pulp-capping material.

To evaluate the multifunctionality of silver-copper co-loaded mesoporous bioactive glass (MBG), with the goal of developing an advanced pulp-capping material. The synthesis of materials was conducted using the sol-gel method, following the approach described in previous studies but with some modifications. The composition included 80 mol% SiO₂, 15 mol% CaO, and 5 mol% P₂O₅, with additional components of 5 mol% silver, 5 mol% copper, or 1 mol% silver combined with 4 mol% copper, designated as Ag5/80S, Cu5/80S, or Ag1Cu4/80S, respectively. Furthermore, crystal phases, surface morphology, and ion-releasing activity were analyzed using X-ray diffraction (XRD), transmission electron microscopy (TEM), and inductively coupled plasma mass spectrometry (ICP-MS), respectively. Human umbilical vein endothelial cells (HUVECs) were employed to assess wound-healing effects, while human dental pulp stem cells (hDPSCs) were utilized to evaluate osteogenic effects. Textural analyses indicated that Ag1Cu4/80S was successfully synthesized using modified procedures, demonstrating comparable ion co-releasing activity. Ag1Cu4/80S exhibited low toxicity and high cell proliferation rates, with a migration rate of 46 %, significantly higher than the <10 % observed in other groups. In terms of osteogenesis, hDPSCs treated with Ag1Cu4/80S displayed enhanced alkaline phosphatase activity, with mineralization levels 1.6-fold greater than those of untreated controls. The synthesis of Ag1Cu4/80S was successfully optimized. This material demonstrated significant wound-healing and comparable osteogenic effects relative to other tested materials, highlighting its potential for dental applications. Ag₁Cu₄/80S demonstrated a comparable effect on osteogenesis, indicating its potential to promote mineralization and suggesting its applicability in dental treatments.

SOX11 silence inhibits atherosclerosis progression in ApoE-deficient mice by alleviating endothelial dysfunction.

SRY-Box Transcription Factor-11 (SOX11) is a transcriptional regulatory factor that plays a crucial role in inflammatory responses. However, its involvement in atherosclerosis (AS), a cardiovascular disease driven by endothelial cell inflammation, remains unknown. This study aims to elucidate the role of SOX11 in AS. The expression of SOX11 was found to be elevated in the aortic tissue of AS mice induced by feeding ApoE-deficient mice a high-fat diet. Knockdown of SOX11 using lentiviral-mediated SOX11-specific shRNA via tail vein injection resulted in a reduction in plaque area and lipid deposition within plaques at the aortic root. Furthermore, silencing SOX11 led to decreased expression of cell adhesion factors Intercellular Cell Adhesion Molecule-1 and Vascular Cell Adhesion Molecule-1, as well as reduced levels of inflammatory factors Interleukin (IL)-6, IL-1β, and chemokine Monocyte Chemotactic Protein-1. In the human umbilical vein endothelial cells (HUVECs) induced by Tumor Necrosis Factor (TNF)-α, increased inflammation was observed at the cellular level, along with enhanced monocyte adhesion. Infection of HUVECs with lentivirus carrying specific shRNA targeting SOX11 inhibited inflammatory response. Mechanistically, chromatin immunoprecipitation (ChIP)-PCR results revealed that SOX11 bound to the promoters of downstream target genes Tumor Necrosis Factor Receptor-Associated Factor-1 (TRAF1), Cluster of Differentiation (CD)40, and CD36, positively regulating their transcription. In conclusion, SOX11 plays a pivotal role in promoting endothelial cell inflammation. Suppression of SOX11 reduces endothelial cell inflammation by inhibiting the transcription of TRAF1, CD40, and CD36, thereby impeding the progression of atherosclerosis.

Porcine pericardial decellularized matrix bilayer patch containing adipose stem cell-derived exosomes for the treatment of diabetic wounds.

Chronic hard-to-heal wounds pose a significant threat to patients' health and quality of life, and their clinical management remains a challenge. Adipose-derived stem cell exosomes (ADSC-exos) have shown promising results in promoting diabetic wound healing. However, effectively enhancing the retention of exosomes in wounds for treatment remains a key issue that needs to be addressed. There is a pressing need to develop new materials or methods to improve the bioavailability of exosomes. Porcine pericardium, an extracellular matrix-rich tissue, is easily obtainable and widely available. Decellularized porcine pericardium removes cellular components while retaining an extracellular matrix that supports cellular growth, making it an ideal raw material for preparing wound dressings. In this study, we developed porcine pericardial decellularized matrix bilayer patches loaded with ADSC-exos, which were transplanted into diabetic mouse skin wounds. Histological and immunohistochemical analyses revealed that these bilayer matrix patches accelerate wound healing by promoting granulation tissue formation, re-epithelialization, stimulating vascularization, and enhancing collagen production. In terms of the underlying biological mechanism, we found that decellularized extracellular matrix bilayer patches loaded with ADSC-exos enhanced the proliferation and migration of human dermal fibroblasts (HDFs) and HaCaT cells in vitro, and promoted tube formation in human umbilical vein endothelial cells (HUVECs). This research demonstrated that the porcine pericardial decellularized matrix is well-suited for exosome delivery and that these bilayer patches hold great potential in promoting diabetic wound healing, providing evidence to support the future clinical application of ADSC-exos.

Senescent retinal pigment epithelial cells promote angiogenesis in choroidal neovascularization via the TAK1/p38 MAPK pathway.

Senescent retinal pigment epithelial cells play a key role in neovascular age-related macular degeneration (nAMD); however, the mechanisms underlying the angiogenic ability of these cells remain unclear. Herein, we investigated the effects of the senescent adult retinal pigment epithelial cell line-19 (ARPE-19) on wound healing, cell migration and survival, and tube formation abilities of human umbilical vein endothelial cells (HUVECs). Additionally, we used Brown Norway rats to establish a laser-induced choroidal neovascularization (CNV) model for further nAMD-related studies. We found that the wound healing, cell migration, and tube formation abilities of HUVECs were significantly enhanced following culture in conditioned media from senescent ARPE-19cells; this was attributed to the activation of the transforming growth factor β-activated kinase 1 (TAK1)/p38 MAPK pathway. Consistently, we found that the TAK1 inhibitors 5Z-7-oxozeaenol and takinib reversed the effects of conditioned media from senescent ARPE-19cells on the wound healing, migration, survival, and tube formation abilities of HUVECs. We further investigated the therapeutic effects of 5Z-7-oxozeaenol on the laser-induced CNV rat model. We found that TAK1 was activated in IB4+ areas in laser-induced CNV lesions; inhibiting the activity of TAK1 using 5Z-7-oxozeaenol significantly alleviated CNV lesion formation and fluorescein leakage in fundus fluorescein angiography and greatly improved a-waves, b-waves, and OP values, as recorded by electroretinography. Thus, senescent RPE cells may promote angiogenesis via the TAK1/p38 MAPK pathway. Further, inhibiting TAK1 expression alleviates pathological neovascularization and improves retinal function in a laser-induced CNV rat model, highlighting the therapeutic potential of this approach for treating nAMD.

Tumor Vascular Normalization by B7-H3 Blockade Augments T Lymphocyte-Mediated Antitumor Immunity.

Triple-negative breast cancer (TNBC), defined by the absence of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2), presents unique clinical challenges and generally predicts a less favorable prognosis. Despite recent advancements in TNBC treatment, a subset of patients remains resistant to immunotherapy. B7-H3, a member of the B7 family of immune checkpoints, is correlated with poor outcomes in various cancers and is distinctively expressed in tumor vasculature, marking it as a potential biomarker for tumor-associated endothelial cells. We found high expression of B7-H3 in the endothelial cells of the postoperative tissue of TNBC patients. Elevated gene expression of CD276 (encoding B7-H3) and PECAM1 (encoding CD31) in TNBC is associated with poor prognosis. Anti-B7-H3 blockade reduces tumor burden and promotes lymphocyte infiltration in a TNBC mouse model. Additionally, anti-B7-H3 blockade promotes tumor vessel normalization and enhances programmed cell death ligand 1 (PD-L1) expression. Synergistic effects were observed when B7-H3 blockade was combined with programmed cell death protein 1 (PD-1) inhibition in the TNBC mouse model. Furthermore, anti-B7-H3 inhibits human umbilical vein endothelial cell (HUVEC) proliferation by suppression of the nuclear factor kappa-B (NF-κB) signaling pathway. Downregulation of B7-H3 expression in HUVECs promotes lymphocyte trans-endothelial migration. These findings suggest that B7-H3 represents a promising therapeutic target for TNBC, and the combination of anti-B7-H3 and anti-PD-1 therapies may have synergetic effects in treating TNBC.

METTL14/IGF2BP2-MEDIATED M6A MODIFICATION OF STEAP1 AGGRAVATES ACUTE LUNG INJURY INDUCED BY SEPSIS.

Background: Acute lung injury (ALI) is a severe complication of sepsis, characterized by inflammation, edema, and injury to alveolar cells, leading to high mortality rates. Septic ALI is a complex disease involving multiple factors and signaling pathways. STEAP family member 1 (STEAP1) has been reported to be upregulated in a sepsis-induced ALI model. However, the role of STEAP1 in the regulation of septic ALI is not yet fully understood. Methods: The study stimulated human pulmonary microvascular endothelial cells (HPMECs) using lipopolysaccharides (LPS) to establish an in vitro ALI model. The study used quantitative real-time polymerase chain reaction to measure mRNA expression, and western blotting assay or immunohistochemistry assay to analyze protein expression. Cell Counting Kit-8 assay was performed to assess cell viability. Flow cytometry was conducted to analyze cell apoptosis. Tube formation assay was used to analyze the tube formation rate of human umbilical vein endothelial cells. Enzyme-linked immunosorbent assays were used to measure the levels of interleukin 1beta and tumor necrosis factor alpha. The levels of Fe 2+ and reactive oxygen species were determined using colorimetric and fluorometric assays, respectively. The glutathione level was also determined using a colorimetric assay. m6A RNA immunoprecipitation assay, dual-luciferase reporter assay, and RNA immunoprecipitation assay were performed to identify the association of STEAP1 with methyltransferase 14, N6-adenosine-methyltransferase noncatalytic subunit (METTL14) and insulin like growth factor 2 mRNA binding protein 2 (IGF2BP2). The transcript half-life of STEAP1 was analyzed by actinomycin D assay. Finally, a rat model of polymicrobial sepsis was established to analyze the effects of STEAP1 knockdown on lung injury in vivo . Results: We found that the mRNA expression levels of STEAP1 and METTL14 were upregulated in the blood of ALI patients induced by sepsis compared to healthy volunteers. LPS treatment increased the protein levels of STEAP1 and METTL14 in HPMECs. STEAP1 depletion attenuated LPS-induced promoting effects on HPMECs' apoptosis, inflammatory response, and ferroptosis, as well as LPS-induced inhibitory effect on tube formation. We also found that METTL14 and IGF2BP2 stabilized STEAP1 mRNA expression through the m6A methylation modification process. Moreover, METTL14 silencing attenuated LPS-induced effects by decreasing STEAP1 expression in HPMECs, and STEAP1 silencing ameliorated cecal ligation and puncture-induced lung injury of mice. Conclusion: METTL14/IGF2BP2-mediated m6A modification of STEAP1 aggravated ALI induced by sepsis. These findings suggest potential therapeutic targets for the treatment of this disease.

Interleukin 29 is a novel antiangiogenic factor in angiogenesis.

Angiogenesis is tightly controlled by growth factors and cytokines in pathophysiological settings. Despite the importance of Interleukin 29 (IL-29), a newly identified cytokine of type III interferon family, its role in angiogenesis remains unknown. We aimed to elucidate IL-29's impact on angiogenesis under both and physiological and pathological conditions. We employed various assays to evaluate IL-29's effect on proliferation, apoptosis, migration and tube formation of human umbilical vein endothelial cells (HUVEC) in vitro. IL-29's angiogenic effect was assessed using mouse aortic rings ex vivo, and oxygen-induced retinopathy (OIR) mouse model in vivo. Signaling pathways possibly involved in IL-29-induced angiogenesis were investigated by Western blot. Finally, IL-29's impact on tube formation was blocked by inhibiting IL-29/interleukin 10 receptor 2 (IL-10R2) binding. IL-29 treatment inhibited endothelial cell migration, tube formation and vessel sprouting, without affecting proliferation or apoptosis. Notably, IL-29 (100ng/ml) attenuated vessel growth in pathological angiogenesis in OIR mice, accompanied by decreased expression of vascular endothelial growth factor (VEGF) and hypoxia inducible factor-1α (HIF-1α). Mechanistically, IL-29 activated Stat3 signaling pathway, and blocking IL-29/IL-10R2 binding remarkably reversed IL-29's anti-angiogenic effect on tube formation. Our findings demonstrated that IL-29, at a relative low concentration, modulates angiogenesis in both physiological and pathological contexts. Targeting IL-29 or its receptor IL-10R2 offers a promising strategy for angiogenesis regulation in various conditions.