ObjectivesEndothelial cell dysfunction during aging is a key driver of vascular aging and related diseases; however, effective strategies to selectively eliminate senescent endothelial cells and restore vascular function remain lacking. FOXO4-DRI, a novel peptide-based intervention, specifically disrupts the interaction between FOXO4 and P53, thereby inducing apoptosis in senescent cells. This study innovatively focuses on the mechanism by which FOXO4-DRI induces apoptosis in senescent endothelial cells, demonstrating that it functions by activating the p53/BCL-2/Caspase-3 signaling pathway to promote selective apoptosis of these cells. FOXO4-DRI significantly improves vascular function and delays vascular aging. These findings not only enrich the molecular understanding of senescent cell clearance but also provide a novel strategy for precise targeting of endothelial cell senescence in therapeutic applications.Materials and methodsThis study aims to analyze the vascular function and aging status of the aorta in naturally aged mice and progeroid model mice following FOXO4-DRI injection. Additionally, it investigates changes in endothelial cell function in senescent endothelial cells induced by oxygen-glucose deprivation (OGD), as well as the protein expression and interaction in the FOXO4-P53 signaling pathway. To assess the impact of FOXO4-DRI on endothelial cell senescence, the senescent endothelial cells were treated with FOXO4-DRI, followed by immunofluorescence and Western blotting experiments.ResultsInjection of FOXO4-DRI in both naturally aged and induced aging mice effectively suppressed aortic aging and improved aortic function. Additionally, we found that FOXO4-DRI alleviates endothelial cell senescence induced by OGD, thereby enhancing endothelial cell function. Through co-immunoprecipitation (CO-IP) experiments, we discovered that FOXO4-DRI prevents the binding of FOXO4 to P53, facilitating the phosphorylated P53 nuclear exclusion, which subsequently trigger BAX and cleaved caspase-3, leading to the apoptosis of senescent cells. Ultimately, this mechanism achieves the goal of inhibiting vascular aging.ConclusionFOXO4-DRI promotes the nuclear export of phosphorylated P53 by inhibiting the binding of FOXO4 to P53 in endothelial cells, thereby facilitating the apoptosis of senescent endothelial cells and alleviating aging.

AIMS:Sepsis-induced cardiomyopathy (SIC) lacks effective targeted therapies. This study investigated whether inhibiting neutrophil infiltration and neutrophil extracellular traps (NETosis) preserves cardiac function and mitigates inflammation and apoptosis in sepsis. METHODS:We performed analyses of gene-expression microarray datasets and single-cell RNA sequencing to define the SIC landscape, and generated cardiac RNA sequencing in LPS-treated versus control mice to validate microarray signatures. In vitro, we used two complementary models, purified NETs applied to HL-1 cardiomyocytes and HL-1-neutrophil coculture to induce NETs, ROS, inflammation, and apoptosis,to evaluate NET-targeting interventions, including CI-amidine, PAD4 inhibition; Sivelestat, neutrophil elastase inhibition; DNase I, NET degradation. To define NET subtype, we verified that PMA-induced NETs were NADPH oxidase-dependent by demonstrating apocynin sensitivity, and we further showed that the mitochondrial ROS scavenger Mito-TEMPO reduced NET-induced ROS in HL-1 cells. In vivo, mice were assigned to control, CLP, or LPS groups; CLP/LPS mice received vehicle or a NET inhibitor. Echocardiography, plasma LDH/CK-MB, IL-1β/6/TNF-α, cardiac NETs (Ly6G/PAD4, MPO/NE IF) and apoptosis (TUNEL, BAX/BCL-2/cleaved caspase-3) were blindly assessed. RESULTS:Datasets-based analyses highlighted immune activation with programmed cell death enrichment, prominently featuring NETosis pathways. Cardiac RNA sequencing in LPS hearts confirmed upregulation of neutrophil/NET-related signatures. Mechanistically, the induced NETs were NADPH oxidase-dependent. In vitro, both purified NETs intervention and HL-1-neutrophil coculture were used to elicit ROS, inflammatory signaling, and apoptosis, and NET-inducing conditions were consistently attenuated by netosis-targeted interventions. CLP and LPS animal model produced early declines in EF/FS, rises in LDH/CK-MB and proinflammatory cytokines, increased cardiac neutrophil infiltration and NET burden, and enhanced cardiomyocyte apoptosis. The neutrophil-targeted interventions reduced cardiac neutrophils and NETs, lowered IL-1β/IL-6/TNF-α, decreased TUNEL positivity and apoptotic signaling, and preserved systolic function. CONCLUSIONS:Our findings indicate that NETosis and neutrophil infiltration are therapeutically actionable contributors in SIC. Strategies that inhibit NET formation, degrade extracellular DNA, or limit neutrophil recruitment were associated with reduced myocardial inflammation and apoptosis and partial preservation of systolic function. These results provide a theoretical basis for early diagnosis, timely intervention, and improved prognosis in SIC.

Atrial fibrillation (AF) is a common arrhythmia encountered in clinical practice, characterized by myocardial fibrosis and atrial remodeling as its primary pathological features, and associated with significantly high mortality and disability rates. Currently, there are no specific pharmacological treatments for AF, and traditional anti-arrhythmic drugs have not achieved the desired efficacy, often resulting in a high incidence of adverse drug reactions. Thus, there is an urgent need for the development of novel anti-AF medications. Berberine, the main active component of Coptis chinensis, has been shown to have antiarrhythmic and anti-heart failure effects. However, its potential to improve atrial fibrosis and remodeling resulting from AF remains largely unexplored. In this study, we used a rapid atrial pacing (RAP) procedure to establish a rabbit model of AF associated with atrial fibrosis. Our objective was to assess the inhibitory effects of berberine on myocardial fibrosis, evaluate its impact on atrial remodeling, and investigate its underlying molecular mechanisms. Our findings indicate that berberine reduces left atrial weight and the area of myocardial fibrosis, inhibits the expression of α-SMA protein in atrial tissue, and decreases the levels of inflammation and oxidative stress. In addition, berberine effectively inhibits atrial remodeling, which may contribute to the prevention of AF. Through transcriptomics, molecular docking, and molecular dynamics simulations, we have tentatively confirmed that berberine may activate the AMPK-PPARα signaling pathway by directly binding to AMPK and PPARα, thereby improving atrial fibrillation.

Diabetic cardiomyopathy (DCM) in type 2 diabetes (T2D) may lead to heart failure and patient death. Fibroblast growth factor 21 (FGF21) is a therapeutic candidate for treating this disease. However, one impediment to its clinical use is its weak ability to activate downstream signaling pathways. In this study, based on our in-depth understanding of the binding properties of fibroblast growth factor receptor 1c (FGFR1c) with paracrine FGF1 and endocrine FGF21, we engineered a novel FGF21 analog named FGF21D2D3. This was achieved by substituting the R96–V106 region of FGF21 (the binding site with the D2–D3 domain of FGFR1c) with the corresponding region from FGF1. Structural characterization and binding affinity tests showed that the analog’s capacity to bind FGFR1c was significantly enhanced compared to wild-type FGF21 (FGF21WT). In a T2D mouse model, we found that FGF21D2D3 had greater potency than FGF21WT in improving hyperlipidemia and DCM. Furthermore, mechanistic studies revealed that FGF21D2D3 more effectively bound FGFR1, activated AMPK, inhibited oxidative stress, and ameliorated DCM. Therefore, our data indicate that FGF21D2D3 is a better substitute for FGF21WT in treating DCM by improving dyslipidemia and directly suppressing oxidative stress via FGFR1–AMPK activation in T2D.

Traditional Chinese medicine (TCM) exerts integrative effects on complex diseases owing to the characteristics of multiple components with multiple targets. However, the syndrome-based system of diagnosis and treatment in TCM can easily lead to bias because of varying medication preferences among physicians, which has been a major challenge in the global acceptance and application of TCM. Therefore, a standardized TCM prescription system needs to be explored to promote its clinical application. In this study, we first developed a gradient weighted disease-target-herbal ingredient-herb network to aid TCM formulation. We tested its efficacy against intracerebral hemorrhage (ICH). First, the top 100 ICH targets in the GeneCards database were screened according to their relevance scores. Then, SymMap and Traditional Chinese Medicine Systems Pharmacology (TCMSP) databases were applied to find out the target-related ingredients and ingredient-containing herbs, respectively. The relevance of the resulting ingredients and herbs to ICH was determined by adding the relevance scores of the corresponding targets. The top five ICH therapeutic herbs were combined to form a tailored TCM prescriptions. The absorbed components in the serum were detected. In a mouse model of ICH, the new prescription exerted multifaceted effects, including improved neurological function, as well as attenuated neuronal damage, cell apoptosis, vascular leakage, and neuroinflammation. These effects matched well with the core pathological changes in ICH. The multi-targets-directed gradient-weighting strategy presents a promising avenue for tailoring precise, multipronged, unbiased, and standardized TCM prescriptions for complex diseases. This study provides a paradigm for advanced achievements-driven modern innovation in TCM concepts.

Loss-of-function variants of low-density lipoprotein receptor-related protein 5 (LRP5) can lead to reduced bone formation, culminating in diminished bone mass. Our previous study reported transcription factor osterix (SP7)-binding sites on the LRP5 promoter and its pivotal role in upregulating LRP5 expression during implant osseointegration. However, the potential role of SP7 in ameliorating LRP5-dependent osteoporosis remained unknown. In this study, we used mice with a conditional knockout (cKO) of LRP5 in mature osteoblasts, which presented decreased osteogenesis. The in vitro experimental results showed that SP7 could promote LRP5 expression, thereby upregulating the osteogenic markers such as alkaline phosphatase (ALP), Runt-related transcription factor 2 (Runx2), and β-catenin (P<0.05). For the in vivo experiment, the SP7 overexpression virus was injected into a bone defect model of LRP5 cKO mice, resulting in increased bone mineral density (BMD) (P<0.001) and volumetric density (bone volume (BV)/total volume (TV)) (P<0.001), and decreased trabecular separation (Tb.Sp) (P<0.05). These data suggested that SP7 could ameliorate bone defect healing in LRP5 cKO mice. Our study provides new insights into potential therapeutic opportunities for ameliorating LRP5-dependent osteoporosis.

Acute respiratory distress syndrome (ARDS) is a severe condition with complex pathogenesis, and emerging evidence highlights the potential role of metabolic factors, though the exact mechanisms are not fully understood. In this study, we used Mendelian randomisation (MR) and multi-omics approaches to investigate the causal relationship between plasma metabolites, immune cell profiles and ARDS risk. MR analysis of 1400 metabolites identified two causal metabolites linked to increased ARDS risk, primarily involved in pantothenate and CoA biosynthesis. Single-cell RNA sequencing of ARDS samples revealed that monocytes exhibited the highest levels of pantothenate synthesis. Intercellular communication and pseudotime analysis suggested that the pantothenate synthesis pathway influenced monocyte differentiation and interactions with other cell types. Gene set enrichment analysis showed that monocytes with high pantothenate synthesis were significantly enriched in phagocytosis-related pathways. Subsequent MR analysis demonstrated that CD33dim HLA DR+ CD11b+%CD33dim HLA DR+ were a risk factor against ARDS. Notably, monocytes with high pantothenate synthesis exhibited decreased expression of antigen presentation markers HLA-DRB5, HLA-DRB1 and HLA-DRA, suggesting that the high pantothenate synthesis monocytes exhibit attenuated antigen presentation and enhanced phagocytic function. Moreover, we developed a diagnostic model using machine learning algorithms. Shapley Additive explanation (SHAP) was leveraged to evaluate the model performance, with CALM2 identified as the most influential feature across the CatBoost and XGBoost models. In summary, this study integrates genetic, multi-omics and machine learning approaches to provide novel insights into the pathogenesis of ARDS and its potential therapeutic strategies targeting monocyte metabolism and function.

Polycaprolactone (PCL) is one of the most widely used three-dimensional (3D) printing materials with excellent biocompatibility and mechanical properties. However, its hydrophobic nature hinders cell adhesion and proliferation. Polydopamine (PDA) has been shown to promote proliferation and induce osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) on polymer surfaces. Despite this, the impact of varying PDA coating thicknesses on the osteogenic differentiation of BMSCs has been minimally explored. In this paper, PCL scaffolds were fabricated using 3D printing technology, and PDA-coated PCL scaffolds (PDA-PCL-0, PDA-PCL-3, PDA-PCL-6, PDA-PCL-24) were prepared by immersing the scaffolds in an aqueous dopamine solution for fixed time points (0, 3, 6, 24 h) under constant shaking. The scaffolds were characterized and subjected to physicochemical performance tests to evaluate their effects on BMSC proliferation, adhesion, and osteogenic differentiation. The results showed that PDA-PCL-6 scaffolds exhibited significant immunomodulatory properties, promoting BMSC proliferation, adhesion, and osteogenic differentiation more effectively than the other groups. In vivo validation experiments, including micro-computed tomography, hematoxylin and eosin staining, Masson staining, and immunohistochemical analysis of bone morphologenetic protein 2 (BMP-2) and type I collagen (COL-I), confirmed that PDA-PCL-6 scaffolds significantly enhanced bone regeneration, histocompatibility, and hemocompatibility compared to uncoated scaffolds at 1, 2, and 3 months postoperation. In conclusion, our results indicate that a PDA coating obtained through 6-h immersion significantly enhances the biocompatibility and osteoinductive properties of PCL scaffolds, providing a promising strategy for bone defect repair.

The importance of macrophages in kidney diseases has been well established; however, the mechanisms underlying the infiltration of macrophages into injured kidneys are not well understood. RGMb is a member of the repulsive guidance molecule (RGM) family. RGMb can be expressed on the cell surface but a large portion of RGMb is localized intracellularly. Among various immune cell types, macrophages express the highest levels of RGMb, but the biological functions of RGMb in macrophages remain largely unknown. We find that RGMb promoted macrophage migration in vitro and that in vivo, RGMb enhanced infiltration of macrophages into injured kidneys and aggravated kidney inflammation and injury in mice. Mechanistically, RGMb bound to TAB1 inside the cell and facilitated the interaction between TRAF6 ubiquitin ligase and TAB1, thereby promoting TRAF6-mediated K63-linked polyubiquitination and phosphorylation of TAK1, followed by increased αTAT1 phosphorylation and α-tubulin acetylation. The resulting changes in the cytoskeleton promoted macrophage migration in vitro and in vivo. Deletion of Rgmb in macrophages markedly reduced TAK1 phosphorylation, αTAT1 phosphorylation, and α-tubulin acetylation and attenuated macrophage infiltration, renal inflammation, tubular injury, and interstitial fibrosis during kidney injury. Our results suggest that macrophage RGMb promotes kidney disease by increasing macrophage infiltration via the TRAF6-TAB1-TAK1/αTAT1/α-tubulin cascade.

The therapy of large defects in peripheral nerve injury (PNI) suffers from several drawbacks, especially the lack of autologous nerve donors. Nerve conduits are considered as a solution for nerve injury treatment, but biocompatibility improvements is still required for conduits prepared with synthetic materials. Cell-derived extracellular matrix (ECM) has drawn attention due to its lower risk of immunogenic response and independence from donor availability. The goal of this study is to coat bone mesenchymal stem cell-derived ECMs on poly(lactic-co-glycolic) acid (PLGA) conduits to enhance their ability to support neural growth and neurite extensions. The ECM-coated conduits have better hydrophilic properties than the pure PLGA conduits. A marked increase on PC12 and RSC96 cells' viability, proliferation and dorsal root ganglion neurite extension was observed. Quantitative PCR analysis exhibited a significant increase in markers for cell proliferation (GAP43), neurite extension (NF-H, MAP2, andβIII-tubulin) and neural function (TREK-1). These results show the potential of ECM-coated PLGA conduits in PNI therapy.

Isoniazid and rifampicin co-therapy are the main causes of anti-tuberculosis drug-induced liver injury (ATB-DILI) and acute liver failure, seriously threatening human health. However, its pathophysiology is not fully elucidated. Growing evidences have shown that fibroblast growth factors (FGFs) play a critical role in diverse aspects of liver pathophysiology. The aim of this study is to investigate the role of FGFs in the pathogenesis of isoniazid (INH) and rifampicin (RIF)-induced liver injury. Through systematic screening, this study finds that hepatic FGF1 expression is significantly downregulated in both mouse model and human patients challenged with INH and RIF. Hepatocyte-specific Fgf1 deficiency exacerbates INH and RIF-induced liver injury resulted from elevated bile acids (BAs) synthases and aberrant BAs accumulation. Conversely, pharmacological administration of the non-mitogenic FGF1 analog - FGF1ΔHBS significantly alleviated INH and RIF-induced liver injury via restoring BAs homeostasis. Mechanically, FGF1 repressed hepatocyte nuclear factor 4α (Hnf4α) transcription via activating FGF receptor 4 (FGFR4)-ERK1/2 signaling pathway, thus reducing BAs synthase. The findings demonstrate hepatic FGF1 functions as a negative regulator of BAs biosynthesis to protect against INH and RIF-induced liver injury via normalizing hepatic BAs homeostasis, providing novel mechanistic insights into the pathogenesis of ATB-DILI and potential therapeutic strategies for treatment of ATB-DILI.

INTRODUCTION:Preeclampsia (PE) is a pregnancy complication that encompasses various pathogenic mechanisms. Shallow implantation of the placenta due to abnormal trophoblast behavior is considered an important mechanism underlying PE; however, its exact etiology remains unclear.METHODS:The expression of OLFML3 in the placenta and important clinical indicators were performed, followed by a correlation analysis. The effect of OLFML3 on the behavior of HTR-8/SVneo cells was examined, and the downstream molecular mechanisms of OLFML3 were investigated in HTR-8/SVneo cells. Additionally, a rat model of PE was generated by adenovirus injection via the tail vein to verify the role of OLFML3.RESULTS:OLFML3 is highly expressed in both syncytiotrophoblasts and cytotrophoblasts and deregulated in preeclamptic placentas. OLFML3 overexpression in HTR-8/SVneo cells promoted cell proliferation, migration, invasion, and impeded apoptosis, and triggered phosphorylation on ser473 of AKT. Conversely, OLFML3 knockdown exerted opposite effects. Furthermore, OLFML3 overexpression ameliorates CoCl2-induced apoptosis of HTR-8/SVneo cells. In a rat model, OLFML3 overexpression alleviates PE-associated maternal symptoms, leading to lower blood pressure, less severe proteinuria, improved fetal growth restriction, as well as upregulation of P-AKT and downregulation of Cleaved caspase3 and Bax.DISCUSSION:OLFML3 may alleviate PE development by inhibiting extravillous trophoblast cell apoptosis through the PI3K/AKT pathway. Our findings indicated that OLFML3 may provide a possible therapeutic target for PE.

Aberrant expression of ATPase sarcoplasmic/endoplasmic retic Ca2+ transporting 2 (ATP2A2) has attracted attention for its pathophysiologic role in pulmonary hypertension (PH). Several miRNAs, including miR-210-5p, have also been reported to be pathogenic factors in PH, but their exact mechanisms remain unknown. This study aimed to elucidate the potential mechanisms of miR-210-5p and ATP2A2 in MCT-induced PH. Eighteen Sprague–Dawley rats were randomly divided into two groups—monoclonal (MCT) group and control group—and then administered MCT (60 mg/kg) and saline, respectively. mPAP, PVR, RVHI, WT

Background and AimsPostoperative atrial fibrillation (POAF) is considered the most prevalent irregular heart rhythm after heart surgery. The cardiac autonomic nervous system significantly affects POAF, and neuropeptide Y (NPY), an abundant neuropeptide in the cardiovascular system, is involved in this autonomic regulation. The current work aimed to examine the potential association of NPY with POAF in individuals administered isolated off-pump coronary artery bypass grafting.MethodsFrom January 1 to May 31, 2020, we examined consecutive cases administered successful isolated off-pump coronary artery bypass grafting with no previously diagnosed atrial fibrillation (AF). Clinical characteristics and plasma samples were collected before surgery. NPY was quantified by enzyme-linked immunosorbent assay (ELISA) in peripheral blood, and POAF cases were identified through a 7-day Holter monitoring.ResultsAmong 120 cases with no previously diagnosed AF, 33 (27.5%) developed POAF during hospitalization. Median NPY levels were markedly elevated in the POAF group in comparison with the sinus rhythm group (31.72 vs. 27.95, P=0.014). Multivariable logistic regression analysis revealed age (OR=1.135, 95%CI 1.054-1.223; p=0.001), left atrial size (OR=1.136, 95%CI 1.004-1.285; p=0.043), and NPY levels in peripheral blood (OR=1.055, 95%CI 1.002-1.111; p=0.041) independently predicted POAF. Additionally, NPY levels were positively correlated with high-frequency (HF) (r=0.2774, p=0.0022) and low-frequency (LF) (r=0.2095, p=0.0217) components of heart rate variability.ConclusionIn summary, this study demonstrates an association between elevated NPY levels in peripheral blood before surgery and POAF occurrence.

Background Acute lung injury (ALI) is a devastating clinical disorder with a high mortality rate, and there is an urgent need for more effective therapies. Fibroblast growth factor 18 (FGF18) has potent anti-inflammatory properties and therefore has become a focus of research for the treatment of lung injury. However, the precise role of FGF18 in the pathological process of ALI and the underlying mechanisms have not been fully elucidated. Methods A mouse model of ALI and human umbilical vein endothelial cells (HUVEC) stimulated with lipopolysaccharide (LPS) was established in vivo and in vitro. AAV-FGF18 and FGF18 proteins were used in C57BL/6J mice and HUVEC, respectively. Vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and p65 protein levels were determined by western blotting or immunofluorescent staining. Afterward, related inhibitors were used to explore the potential mechanism by which FGF18 relieves inflammation. Results In this study, we found that FGF18 was significantly upregulated in LPS-induced ALI mouse lung tissues and LPS-stimulated HUVECs. Furthermore, our studies demonstrated that overexpressing FGF18 in the lung or HUVEC could significantly alleviate LPS-induced lung injury and inhibit vascular leakage. Conclusions Mechanically, FGF18 treatment dramatically inhibited the NF-κB signaling pathway both in vivo and in vitro. In conclusion, these results indicate that FGF18 attenuates lung injury, at least partially, via the NF-κB signaling pathway and therefore may be a potential therapeutic target for ALI.

Acute kidney injury (AKI) is a devastating clinical condition characterized by an abrupt loss of renal function. The pathophysiology of AKI involves diverse processes and elements, of which survival and regeneration have been established to be significant hallmarks. And early studies have confirmed the fundamental role of FGFs in the regulation of AKI pathology, although the association between FGF18 and AKI still remains elusive. Our study demonstrates a substantial up-regulation of FGF18 in the renal tubules of mice subjected to ischemia. Notably, targeted overexpression of FGF18 effectively mitigates the impairment of kidney function induced by AKI. Mechanistically, FGF18 facilitates cell proliferation and anti-apoptosis in RTECs by enhancing the expression of YAP and facilitating its translocation to the nucleus. Aside from that, we also discovered that the substantial expression of FGF18 under ischemic conditions is HIF-1α dependent. This study aims to uncover the inherent mechanism behind the beneficial effects of FGF18 in attenuating AKI. By doing so, it aims to offer novel insights into the development of therapeutic strategies for AKI.

The skin barrier is essential to prevent pathogenic invasion. When injury occurs, multiple biological pathways are promptly activated and wound repair processes are triggered. The effective healing of wounds is essential for survival, and dysfunction could result from aberrant wound repair. Preparation of many hydrogels, which involve the addition of growth/cell factors or mimic extracellular matrix (ECM) components, has not resulted in significant advances in tissue recovery. ECM contains a large number of biologically active molecules that activate a variety of cellular transduction pathways, which are essential for wound repair. Here, this work prepares hyaluronic acid-dopamine-thiourea (HA-DA-NCSN) hydrogels exhibiting ultrafast gelation in situ, following the methods of Xu et al., and subsequently designs a hydrogel containing ECM particles. In addition, the loaded ECM material, specifically decellularized ECM material, not only enhances the strength of the hydrogel network, but also delivers bioactive substances that make it a suitable platform for skin wound repair. The ECM hydrogel has great potential as an efficient bioactive wound dressing. This research suggests that this strategy is likely to improve skin wound closure in rat skin wound models.