Acute inflammation, when unresolved, can lead to complications that impair tissue repair and therapeutic outcomes. In this study, we employed a model of lipopolysaccharide (LPS)-induced acute subcutaneous abdominal inflammation in mice to investigate the modulatory effects of elastic compression. LPS administration elicited a robust inflammatory response, characterized by increased leukocyte infiltration, edema, and upregulation of pro-inflammatory mediators. Elastic compression significantly attenuated this response, reducing leukocyte counts in subcutaneous lavage, histological inflammatory infiltrates, and the expression of key pro-inflammatory genes and proteins, including NF-κB, IL-1β, and TNF-α, at both 24 and 72 hours post-induction. Mechanistically, these effects may result from the compressive force altering microvascular dynamics and modulating macrophage polarization and mechanotransduction pathways, including TLR4 and integrin signaling. Additionally, compression preserved redox homeostasis, as indicated by stable oxidative stress markers and antioxidant responses. To our knowledge, this is the first study to demonstrate that elastic compression modulates inflammation at molecular, cellular, and tissue levels in an acute inflammation model. These findings support the therapeutic potential of elastic compression as a non-pharmacological strategy for managing acute inflammation, with possible applications in postoperative care, traumatic edema, and other soft tissue inflammatory conditions. Further translational and clinical studies are warranted to validate these outcomes and guide evidence-based application protocols. This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .
Inflammation and fibrinolytic imbalance, which are frequently caused by oxidative stress and increased pro-inflammatory mediators, are major factors in the onset and progression of chronic illnesses. The study evaluated the phytochemical profile of Urena lobata L. leaf extracts using qualitative chemical tests and spectroscopic analysis, including FT-IR and MC-MS profiling. The pharmacological assessments were performed by antioxidant, anti-inflammatory, and thrombolytic capabilities of extracts using various in vitro assay methods along with in silico approaches, including molecular docking and ADMET-based pharmacokinetic assessments. The methanolic extract of Urena lobata L. (ULM) was fractionated using n-hexane, chloroform, ethyl acetate, and water to obtain the n-hexane (ULH), chloroform (ULC), ethyl acetate (ULE), and aqueous (ULW) fractions. Phytochemical analysis exhibited the presence of flavonoids, steroids, terpenoids, alkaloids, glycosides, and phenols in various fractions. FTIR analysis revealed the presence of a variety of groups, including aliphatic, aromatic, amide, nitrate, methylene, alcohols, phenols, etc. GC-MS study revealed significant bioactive chemicals, including β-amyrenonol acetate derivative, 14-deoxy-12-hydroxyandrographolide, 9,12-octadecadienoic acid, and N-(5-methyl-1,3-thiazol-2-yl)-2-phenylacetamide. In in vitro antioxidant assay, ULE had the highest antioxidant activity among the investment fractions, with IC50 values of 1.17 µg/mL (DPPH) and 2.60 µg/mL (hydroxyl radical), followed by ULM, ULC, and ULH fractions. In vitro anti-inflammatory studies indicated that the ULE efficiently stabilized human red blood cell membranes (~ 91.57%), comparable to that of the standard medication diclofenac. Additionally, thrombolytic assays revealed considerable clot lysis for ULE (28.40%) and ULM (28.26%) fractions. Molecular docking revealed that β-amyrenonol acetate derivative had a stronger binding affinity to COX-2 (-10.1 kcal/mol) than conventional diclofenac and plasminogen (-9.8 kcal/mol), whereas other elements showed synergistic interactions. ADMET calculation validated favorable pharmacokinetic features and low toxicity risk. Notably, this study presents the first in silico investigation of U. lobata L. phytochemicals as potential thrombolytic agents. Overall, the ethyl acetate fraction of U. lobata L. has substantial antioxidant, anti-inflammatory, and thrombolytic properties, highlighting its promise as a multifunctional therapeutic agent and justifying additional bioassay-guided isolation, mechanistic research, and clinical testing.
Chronic ethanol exposure activates inflammatory signaling pathways and inflicts hepatocellular damage, leading to alcohol-associated liver diseases (ALDs). ALD is one of the major causes of global burden, yet there are no FDA-approved treatment options available. This study evaluates the hepatoprotective effects of short-chain fatty acids (SCFAs), mainly sodium acetate (NaA) and sodium butyrate (NaB), against ethanol-induced inflammation and oxidative stress in both in vitro (Buffalo Rat Liver-3A [BRL3A]) and in vivo (male Wistar rats) models. The treatment of NaA and NaB and their combination was given to the cell lines where maximum viability was observed at concentrations of 1.5 mM, 5 mM, and 0.1 mM + 1 mM, respectively. Additionally, reactive oxygen species (ROS) and nuclear morphology were assessed by fluorescent staining. For in vivo samples, the hepatic injury was analyzed by serum biochemical markers. Furthermore, hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) staining were employed, which provided structural and immunological alterations in hepatic tissue. RT-qPCR profiled the expression levels of various pro-inflammatory and anti-inflammatory cytokines, as well as cytochrome P450 E1 (CYP2E1) and antioxidative stress markers. Moreover, enzyme-linked immunosorbent assay (ELISA) quantified the essential protein targets such as TNF-α, MCP-1, IL-1β, IL-6, HO-1, and Nrf2. The administration of NaA, NaB, and their combination resulted in reduced ROS levels and expression of pro-inflammatory cytokines, preserved nuclear integrity, and neutrophil infiltration. These findings were further confirmed by in silico analysis and conserved amino acid interactions, and the affinities of NaA and NaB for TNF-α and MCP-1 were observed as compared to established inhibitors or activators. This study is the first demonstration to report the synergistic effects of NaA and NaB on the feedback loop of the nuclear factor kappa B (NF-κB) signaling pathway, suggesting their potential as promising therapeutic candidates for alleviating alcohol-induced hepatic damage.
Cardiometabolic diseases, including diabetes mellitus, are complicated by vascular disease, a major driver of morbidity and mortality. Although hyperglycaemia contributes to vascular dysfunction, it does not fully explain the vascular complications observed in patients. Chronic low-grade inflammation and persistent release of pro-inflammatory cytokines as interleukin-1β (IL-1β) are increasingly recognized as central mediators of diabetic vasculopathy. However, the mechanisms by which elevated glucose amplifies inflammatory signalling and vascular dysfunction, and their pharmacological modulation, remain incompletely understood. We investigated the interplay between IL and 1β and high glucose in human aortic smooth muscle cells (HASMC) and its impact on NLRP3 inflammasome activation, cellular metabolism and small extracellular vesicles (sEV)-mediated intercellular communication. IL-1β induced NLRP3 inflammasome activation and a metabolic reprogramming characterized not only by a glycolytic shift, but also by activation of the pentose phosphate pathway and NADPH oxidase. IL-1β promoted the release of sEV enriched in inflammasome components, particularly caspase-1, which propagated inflammation and senescence in recipient vascular cells. High glucose alone had no effect but potentiated IL-1β-induced responses. Pharmacologically, blockade of IL-1R with anakinra prevented inflammasome activation, metabolic reprogramming and sEV release. Moreover, both anakinra and the NLRP3 inhibitor MCC950 impeded, at different levels, the potentiating effect of high glucose on IL-1β-driven responses, reinforcing the relevance of targeting the IL-1β-NLRP3 autoinflammatory axis. These findings reveal that high glucose potentiates IL-1β-driven vascular inflammation by altering bioenergetic flexibility and sEV signalling in human vascular cells, providing novel mechanistic insight into how IL-1β-targeted therapies may mitigate vascular complications in cardiometabolic disorders as diabetes.
Metabolic dysfunction-associated steatohepatitis (MASH) represents a severe form of metabolic dysfunction-associated steatotic liver disease (MASLD), largely due to metabolic dysregulation and sustained liver inflammation. TNF-α plays a pivotal role in MASH pathogenesis by inducing cell death, inflammation, and decreased insulin sensitivity. In this study we investigated the effects of TNF-α in human precision-cut liver slices (PCLS) under healthy or steatotic conditions, to provide insights into MASH pathogenesis. PCLS were prepared from human liver tissue and cultured in control (WEGG) and hyper-nutritive (GFIPO) mediums with or without TNF-α (50 ng/mL) for 96 h. Viability was assessed via ATP content, lipid accumulation by triglyceride (TG) assay, and transcriptomic changes through Next-Generation Sequencing. The protein levels of cytokines, chemokines, and fibrotic mediators released from PCLS were quantified using Luminex assay and ELISA. TNF-α significantly altered the transcriptional profiles in PCLS, inducing pro-inflammatory and pro-fibrotic signaling, and downregulating lipid metabolic processes in both WEGG and GFIPO media. TNF-α showed a trend in elevating intracellular TG in both conditions, albeit not statistically significant. On protein levels, TNF-α supplementation to WEGG medium induced the expression of IL8, CCL2, CCL19, PDGF-AB/BB, TGF-α, and MMP9. GFIPO medium alone induced inflammatory and fibrotic responses indicated by elevated levels of IL8, CCL2, and Pro-collagen 1A1. GFIPO medium with TNF-α supplementation further exacerbated the inflammatory and fibrotic responses, characterized by increased release of cytokines and growth factors. This human PCLS model effectively demonstrated the co-occurrence of key features of MASH, such as steatosis, inflammation, and fibrosis, highlighting the impact of metabolic stress and inflammatory cytokine TNF-α on these disease characteristics, and the potential of the PCLS model in exploring mechanism of MASH progression.
The systemic immune-inflammation index (SII) is a simple parameter that has been used to predict adverse outcomes in different clinical settings. The prognostic value of SII, an innovative biomarker of inflammation, in non-valvular atrial fibrillation (NVAF) patients has not been assessed. The aim of this study was to evaluate the prognostic value of SII in NVAF patients who were under direct oral anticoagulant (DOAC) medication. A total of 467 patients with NVAF treated with DOACs from 2018 to 2021 were retrospectively enrolled in this cohort study. Demographics, medication, and outcome data were collected for each patient. SII was calculated from complete blood counts as the ratio of neutrophil-to-lymphocyte count multiplied by the platelet count. The primary outcome was major adverse events (MAEs), which include major bleeding, stroke, and death. Logistic regression analyses showed that the SII score was an important predictor of outcomes in NVAF patients. The SII may be a successful, non-invasive, and cost-effective parameter for predicting bleeding and stroke in patients with NVAF. It may be used as a useful risk stratification tool for these patients.
One of the major challenges in the treatment of inflammatory bowel disease (IBD) is the development of an oral drug delivery system that can provide extended GI residence time. In this work, a nanoformulation formed by Rebamipide (Re) encapsulated into mucin DEAE-dextran armored nanoparticles (M@D@Re-SCNPs) is introduced, which utilizes the core of caprylic acid (C)/6-O-stearoyl ascorbic acid (S). These M@D@Re-SCNPs were fabricated to provide both gastro-resistant and mucoadhesion properties of the nanoparticles. The M@D@Re-SCNPs showed pH-independent structural integrity in the gastric environment and proved to have a good mucosal binding capacity in the colon for long intestinal retention of up to 48 h. In terms of therapeutic efficacy, post-oral administration of M@D@Re-SCNPs strongly reduce the colon inflammatory status and the expression of inflammatory biomarkers like nuclear factor kappa B (NF-κB), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS) and phosphoinositide 3-kinase (PI3K) when compared with free rebamipide. The trapped rebamipide inhibits the production of pro-inflammatory mediators and promotes the production of mucin 2 (Muc2) protein, which promotes colon mucosal regeneration and goblet cell viability. Based on the necessity of controlling drug release and regulation of drug action for complete IBD treatment, we believe that the M@D@Re-SCNPs are a promising drug delivery system for UC treatment.
Cardiac aging is a fundamental contributor to heart failure, arrhythmias, impaired stress tolerance, and reduced cardiovascular resilience in the elderly. While intrinsic myocardial aging has been widely examined, the systemic mechanisms driving age-related cardiac decline are not fully understood. Circulating extracellular vesicles (EVs) have emerged as key mediators of intercellular and inter-organ communication, transferring proteins, lipids, and nucleic acids that modify the phenotypes of recipient cells. Growing evidence indicates that EVs originating from cardiovascular cells as well as distant organs, including the liver, kidney, adipose tissue, lung, and intestine, participate in cardiac aging by influencing senescence-associated signaling, chronic inflammation, mitochondrial and metabolic dysfunction, fibrosis, microvascular remodeling, and contractile function. In this review, we outline the biological basis of circulating EVs in cardiac aging, describe their cellular and inter-organ sources, and examine how they promote progressive myocardial remodeling. We also consider their potential as biomarkers for cardiac aging and age-related heart disease, along with emerging therapeutic strategies such as blocking pathogenic EV signals, using reparative or engineered EVs, and implementing systemic interventions that modulate EV-mediated communication. Key translational challenges are highlighted, including EV heterogeneity, low tissue specificity, methodological inconsistencies, and the need to distinguish physiological cardiac aging from accelerated aging and age-related cardiovascular disease, as EV profiles may reflect systemic comorbidities rather than intrinsic cardiac aging. A deeper understanding of systemic EV crosstalk may offer new insights into the aging heart and support more precise methods for risk stratification, monitoring, and intervention.
Diabetic neuropathy (DN) is a prevalent microvascular complication of diabetes mellitus, characterized by hyperalgesia and allodynia that severely impair quality of life. Current treatment approaches do not provide adequate relief, largely due to the multifactorial nature of disease pathogenesis. Growing evidence indicates that dysregulation of multiple ion channel families is a central mechanism underlying sensory neuron hyperexcitability and chronic pain in DN. This review comprehensively discusses the roles of major ion channel families, including voltage-gated sodium (Naᵥ), calcium (Caᵥ), and potassium (Kᵥ) channels, transient receptor potential (TRP) channels, purinergic receptors (P2X/P2Y), and mechanosensitive PIEZO (PIEZO 1 and PIEZO 2) channels, in sensory transmission and pain modulation. Their dysregulation, induced by chronic hyperglycemia and oxidative stress, promotes ectopic firing, altered calcium homeostasis, and glial activation, sustaining nociceptive hypersensitivity. The review further evaluates current and emerging ion channel-targeted therapeutic approaches, highlighting mechanistic insights, translational challenges, and future research directions. Recent research highlights multi-target and combination strategies, such as Naᵥ1.8 inhibition with KCNQ activation or concurrent blockade of TRPV1 and P2X3, as promising avenues offering synergistic analgesic benefits and disease-modifying potential. Advances in nanocarrier-based delivery, gene modulation, and patient-specific electrophysiological profiling further enhance translational prospects. Ultimately, the therapeutic landscape of PDN is shifting from single-channel blockade toward integrated approaches that modulate excitability, inflammation, and metabolic stress concurrently. Ion channels thus represent not only crucial mediators of PDN pathophysiology but also versatile therapeutic targets whose selective and combinatorial modulation may transform the management of diabetic neuropathic pain. See also the graphical abstract(Fig. 1).
Cancer-associated fibroblasts (CAFs) are critical components of the tumor microenvironment in endometrial cancer (EC). Primary isolation and culture of CAFs serve as the standard method for investigating their biological roles. However, it remains unclear whether prolonged in vitro culture alters the intrinsic properties of CAFs, potentially affecting experimental reproducibility. This study aims to evaluate the phenotypic and transcriptomic evolution of primary EC-derived CAFs across different passages. CAFs were isolated from human EC tissues using enzymatic digestion and differential adherence. Cells at passages 1 (P1), 5 (P5), and 10 (P10) were compared. Biological phenotypes, including proliferation, migration, apoptosis, senescence, and cytokine secretion, were assessed. The effects of CAF-conditioned medium (CM) on the proliferation and invasion of EC cell lines (ISHIKAWA and HEC-1 A) were evaluated. Furthermore, RNA sequencing (RNA-seq) was performed to uncover molecular alterations, followed by functional enrichment and subtype analyses. Additional validation experiments, including ROS detection, NF-κB activation analysis, H₂O₂-induced oxidative stress modeling, and NAC or TPCA-1 intervention, were conducted to investigate the mechanism underlying passage-associated inflammatory drift. CAFs maintained a consistent spindle-shaped morphology and expression of specific markers (Vimentin/FAP) from P1 to P10. Functional assays demonstrated no significant differences in proliferation, migration, or senescence rates across passages. Crucially, CAFs at all passages exhibited comparable efficacy in promoting the proliferation, migration, and invasion of EC cells. However, transcriptomic profiling revealed profound reprogramming, with 1,300 core genes differentially expressed in P5/P10 compared to P1. While pathways related to extracellular matrix remodeling (myCAF signature) remained stable, establishing the molecular basis for sustained invasion support, passaged CAFs progressively exhibited reduced antigen-presenting signatures and acquired a pro-inflammatory iCAF-like phenotype. Mechanistically, late-passage CAFs showed increased ROS accumulation, enhanced P65 phosphorylation and nuclear translocation, and upregulated inflammatory mediators, including IL6, CXCL1, CXCL2, and CCL2. H₂O₂ treatment partially mimicked this inflammatory phenotype in P1 CAFs, whereas NAC or TPCA-1 attenuated inflammatory activation in P10 CAFs. Primary endometrial CAFs retain core tumor-supporting phenotypes and myofibroblastic characteristics up to passage 10, validating their utility as reliable in vitro models for invasion and metastasis studies. However, high-passage CAFs exhibit a culture-associated inflammatory transcriptional drift driven, at least in part, by ROS/NF-κB activation, together with reduced immunogenic features. These findings provide essential guidelines for the standardization of CAF-based research models and caution against using high-passage CAFs for immune-interaction or inflammation-related studies.
Objective: To explore the role and molecular mechanism of peroxiredoxin 1 (PRDX1) in hypertension-induced endothelial dysfunction. Methods: (1) Bioinformatics analysis: A total of 40 C57BL/6J mice aged 8-10 weeks (20-25 g) were randomly divided into the saline group and angiotensin Ⅱ (AngⅡ, 0.8 mg·kg⁻¹·d⁻¹) group, with 20 mice in each group. After 4 consecutive weeks of intervention, mice were sacrificed, and thoracic aortic tissues were collected for transcriptome sequencing. Gene Ontology functional annotation and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were performed on differentially expressed genes. (2) Cell experiments: Human umbilical vein endothelial cells (HUVECs) were divided into the control group (endothelial cell culture medium) and the AngⅡ intervention group (medium containing 10-⁶ mol/L AngⅡ). Wound healing assay, cell adhesion assay, and Transwell assay were used to assess cell migration and adhesion. Lentiviral or small interfering RNA (siRNA) transfection was performed to achieve PRDX1 overexpression and knockdown, respectively. The overexpression experiment was divided into the LV-NC (negative control lentivirus) group, Ang Ⅱ+LV-NC group, LV-PRDX1 (PRDX1 overexpression lentivirus) group and Ang Ⅱ+LV-PRDX1 group. The knockdown experiment was divided into the NC-siRNA (negative control siRNA) group, si-PRDX1 group, NC-siRNA+rapamycin (50 nmol/L) group and si-PRDX1+rapamycin group. Immunofluorescence staining was applied to detect intracellular reactive oxygen species level. Quantitative reverse transcription-polymerase chain reaction was used to detect the mRNA expression levels of PRDX1 and mammalian target of rapamycin (mTOR). Western blot was adopted to determine the total protein and phosphorylation levels of PRDX1, mTOR, p70 ribosomal S6 kinase (p70S6K) 1 and endothelial nitric oxide synthase (eNOS). Co-immunoprecipitation assay was used to verify the protein interaction between PRDX1 and mTOR. Nitrate reductase method was used to measure cellular nitric oxide (NO) content. (3) Animal experiments: Forty C57BL/6J mice aged 8-10 weeks (20-25 g) were used to construct the PRDX1 overexpression model via adeno-associated virus serotype 9 (AAV9) vector. Mice were assigned into 4 groups with 10 animals per group: saline+AAV9-GFP (empty vector) group, saline+AAV9-PRDX1 (recombinant virus) group, AngⅡ+AAV9-GFP group, and AngⅡ+AAV9-PRDX1 group. Systolic blood pressure and diastolic blood pressure of mice in each group were dynamically monitored at day 0, 7, 14, 21 and 28 after modeling. Plasma NO level was detected by the nitrate reductase method. After sacrifice, isolated thoracic aortic tissues were subjected to morphological and pathological staining analysis, and a microvascular tension measurement system was used to evaluate the acetylcholine-mediated endothelium-dependent vasodilation function. Results: (1) Bioinformatics analysis: Transcriptome sequencing revealed that numerous differentially expressed genes were identified in the thoracic aorta of mice in the AngⅡ group compared with the saline group. These genes were mainly enriched in biological processes closely associated with oxidative stress, such as reactive oxygen species metabolism and oxidative phosphorylation regulation. (2) Cell experiments: Compared with the control group, HUVECs in the AngⅡ intervention group presented decreased protein and mRNA levels of PRDX1, as well as elevated phosphorylation levels of mTOR and p70S6K1 (all P<0.05). Compared with the LV-NC group, the LV-PRDX1 group showed higher PRDX1 mRNA expression, lower reactive oxygen species levels, enhanced cell migration and adhesion capacities, and increased NO content (all P<0.05). In contrast with the AngⅡ+LV-NC group, the AngⅡ+LV-PRDX1 group exhibited reduced phosphorylation levels of mTOR and p70S6K1 and increased eNOS phosphorylation level (all P<0.05). In addition, relative to the NC-siRNA group, the si-PRDX1 group had higher reactive oxygen species levels and elevated phosphorylation of mTOR and p70S6K1, accompanied by decreased NO content, reduced eNOS phosphorylation, and weakened cell migration and adhesion abilities (all P<0.05). Compared with the si-PRDX1 group, the above abnormal changes were partially reversed in the si-PRDX1+rapamycin group (all P<0.05). Co-immunoprecipitation assay confirmed a protein interaction between PRDX1 and mTOR. (3) Animal experiments: In comparison with the saline+AAV9-GFP group, the AngⅡ+AAV9-GFP group had higher systolic and diastolic blood pressure, lower plasma NO level, thicker thoracic aortic media, increased collagen deposition, disordered arrangement of elastic fibers, and impaired endothelium-dependent vasodilation in response to acetylcholine (all P<0.05). Notably, the AngⅡ+AAV9-PRDX1 group showed lower systolic and diastolic blood pressure, alleviated pathological damage of the thoracic aorta, improved endothelium-dependent vasodilation function, and higher plasma NO level than the AngⅡ+AAV9-GFP group (all P<0.05). Conclusion: PRDX1 can inhibit the excessive activation of the mTOR/p70S6K signaling pathway by scavenging reactive oxygen species and promoting NO production, thereby regulating eNOS activity and ameliorating endothelial dysfunction and vascular injury under hypertensive conditions. Targeted regulation of the PRDX1/ROS/mTOR/p70S6K signaling axis is expected to provide a novel therapeutic target and intervention strategy for the prevention and treatment of hypertensive vascular diseases. 目的： 探讨过氧化物氧化还原蛋白1（PRDX1）在高血压内皮功能障碍中的作用和分子机制。 方法： （1）生物信息学分析：选取8~10周龄C57BL/6J小鼠（体重20~25 g）40只，随机分为生理盐水组和血管紧张素Ⅱ（AngⅡ，0.8 mg·kg⁻¹·d⁻¹）组，每组20只，连续干预4周后处死小鼠并取胸主动脉组织进行转录组测序，对差异基因进行基因本体功能注释及京都基因与基因组百科全书通路富集分析。（2）细胞实验：选取人脐静脉内皮细胞（HUVECs），分为对照组（内皮细胞培养基）和AngⅡ干预组（含10-6mol/L AngⅡ培养基），采用划痕实验、黏附实验和Transwell实验检测细胞迁移、黏附能力。对HUVECs分别转染慢病毒或小干扰RNA（siRNA）实现PRDX1的过表达与敲低：过表达实验分为LV-NC（阴性对照慢病毒）组、AngⅡ+LV-NC组、LV-PRDX1（PRDX1过表达慢病毒）组和AngⅡ+LV-PRDX1组；敲低实验分为NC-siRNA（阴性对照siRNA）组、si-PRDX1组、NC-siRNA+雷帕霉素（50 nmol/L）组和si-PRDX1+雷帕霉素组；采用免疫荧光染色检测细胞活性氧水平，采用定量逆转录聚合酶链式反应检测PRDX1、哺乳动物雷帕霉素靶蛋白（mTOR）等信使RNA（mRNA）表达水平，Western blot法检测PRDX1、mTOR、p70核糖体S6激酶（p70S6K）1、内皮型一氧化氮合酶（eNOS）总蛋白与磷酸化水平，采用免疫共沉淀实验检测PRDX1与mTOR的蛋白相互作用，采用硝酸还原酶法检测细胞一氧化氮（NO）含量。（3）动物实验：选取8~10周龄C57BL/6J小鼠（体重20~25 g）40只，采用腺相关病毒血清型9（AAV9）载体构建PRDX1基因过表达模型，共设4组，每组10只：生理盐水+AAV9-GFP（空载病毒）组、生理盐水+AAV9-PRDX1（重组病毒）组、AngⅡ+AAV9-GFP组、AngⅡ+AAV9-PRDX1组。于造模后第0、7、14、21、28天动态监测各组小鼠尾动脉收缩压与舒张压，采用硝酸还原酶法检测小鼠血浆NO水平；处死小鼠后对离体主动脉进行组织形态学及病理学染色分析，并采用微血管张力测定系统评估血管对乙酰胆碱的内皮依赖性舒张功能。 结果： （1）生物信息学分析：转录组测序结果显示，与生理盐水组相比，AngⅡ组小鼠胸主动脉组织中存在大量差异表达基因，主要富集于活性氧代谢、氧化磷酸化调控等与氧化应激密切相关的生物学过程。（2）细胞实验：与对照组相比，AngⅡ干预组HUVECs中PRDX1蛋白及mRNA水平均较低，mTOR及p70S6K1磷酸化水平较高（P均<0.05）。与LV-NC组相比，LV-PRDX1组HUVECs中PRDX1 mRNA表达水平较高、活性氧水平较低，细胞迁移和黏附能力均较强，NO含量也较高（P均<0.05）；与AngⅡ+LV-NC组相比，AngⅡ+LV-PRDX1组HUVECs中mTOR和p70S6K1磷酸化水平较低，eNOS磷酸化水平较高（P均<0.05）。此外，与NC-siRNA组相比，si-PRDX1组HUVECs中活性氧水平、mTOR及p70S6K1磷酸化水平较高，NO含量、eNOS磷酸化水平较低，细胞迁移和黏附能力较弱（P均<0.05）；与si-PRDX1组相比，si-PRDX1+雷帕霉素组HUVECs上述改变均得到部分逆转（均P<0.05）。免疫共沉淀实验表明，PRDX1与mTOR存在蛋白相互作用。（3）动物实验：与生理盐水+AAV9-GFP组相比，AngⅡ+AAV9-GFP组小鼠收缩压和舒张压较高，血浆NO水平较低，胸主动脉中膜较厚、胶原沉积增加、弹性纤维排列紊乱，对乙酰胆碱的内皮依赖性舒张反应较弱（P均<0.05）；而AngⅡ+AAV9-PRDX1组小鼠收缩压和舒张压均低于AngⅡ+AAV9-GFP组，上述胸主动脉病理损伤程度较轻，同时对乙酰胆碱的内皮依赖性舒张反应较强，血浆NO水平较高（P均<0.05）。 结论： PRDX1可通过清除活性氧、促进NO生成，抑制mTOR/p70S6K信号通路过度激活，进而调控eNOS活性，改善高血压状态下的内皮功能障碍与血管损伤；靶向调控PRDX1/活性氧/mTOR/p70S6K信号轴有望为高血压血管病变的防治提供新靶点与干预策略。.
The Socioemotional Selectivity Theory of aging posits that the narrowing of social networks with age may provide emotional benefits. It is unknown whether these benefits extend to measures of physical health, such as markers of inflammation. In a longitudinal midlife cohort, we (1) characterized age-related changes in social networks and inflammation, (2) tested associations between changes in social network composition and changes in inflammation, and (3) explored whether changes in positive affect or social support mediated these associations. At waves 1 and 2, ~15 years later, participants from the Adult Health and Behavior study (N=404, 42% male, 89% white, W1 mean age=45.5 years) self-reported their social network composition, positive affect, and social support and provided blood samples to assess IL-6, CRP, TNF-α, and suPAR. Social network composition was characterized using a ratio of the number of close to peripheral members. The social network ratio increased over time and close relationships were more likely to be retained than peripheral (p=.007). Inflammatory markers IL-6, CRP, and suPAR increased over time, but TNF-α slightly decreased (ps <.027). An increase in the social network ratio, indicating more narrowing, was associated with lower levels of TNF-α at W2 (b=-.019, SE=.008, p=.013) and retaining more peripheral relationships was associated with higher levels of CRP (b=.015, SE=.006, p=.015), adjusting for W1 inflammation. Positive affect and social support did not operate as mediators. Our findings provide initial support that network narrowing may be physiologically beneficial in the transition from midlife to late adulthood.
Ferroptosis and macrophage activation are key contributors to the development of acute kidney injury (AKI). Ferroptosis is accompanied by metabolic reprogramming and the release of soluble mediators, including metabolites, cytokines, and extracellular signals, which can propagate tissue damage and modulate immune responses. However, the metabolic profile of ferroptotic tubular epithelial cells and its impact on the immune microenvironment during ischemia-reperfusion injury (IRI) remains largely unexplored. Using untargeted metabolomics, we found that ferroptotic cells secreted abnormally elevated levels of methylmalonic acid (MMA), and investigated the physiological role of MMA in acute kidney injury in mice. Furthermore, through transcriptomics and Western blotting, we explored the mechanism by which the ferroptosis-associated metabolite MMA promotes macrophage polarization. Here, untargeted metabolomics revealed a distinct metabolic secretome of ferroptotic tubular epithelial cells, with the level of MMA markedly elevated after IRI. Mechanistic studies demonstrated that MMA activated the PI3K/ Akt /NF-κB pathway in macrophages, driving M1 polarization and increasing the secretion of proinflammatory cytokines such as IL-6 and TNF-α, ultimately exacerbating acute kidney injury. These findings reveal the mechanism of metabolite-immune crosstalk in AKI, and suggest that targeting the ferroptosis-macrophage axis may represent a therapeutic strategy to disrupt the vicious cycle of inflammation and tissue injury.
Metabolic dysfunction-associated steatotic liver disease (MASLD) is increasingly prevalent. Life's Crucial 9 (LC9) combines cardiometabolic and mental health metrics linked to MASLD pathogenesis, but its association with liver fibrosis and prognosis in MASLD remains unknown. Using National Health and Nutrition Examination Survey (NHANES) data, we performed a prospective cohort analysis for mortality (2005-2016, n = 2524) and a cross-sectional analysis for liver fibrosis (2017-2018, n = 1277). Significant fibrosis was defined as liver stiffness measurement (LSM) ≥8.0 kPa by vibration-controlled transient elastography. Systemic inflammation was assessed using the systemic immune-inflammation index (SII) and pan-immune-inflammation value (PIV). Multivariable regression and mediation analyses were conducted. Higher LC9 scores were associated with lower liver stiffness (β = -0.07, 95% CI: -0.12 to -0.01). Each 1-point increase in LC9 reduced fibrosis risk by 5% (OR = 0.95), with the highest quartile showing the lowest risk (OR = 0.21). Over a median follow-up of 98 months, each 1-point LC9 increase was associated with a 3% reduction in all-cause mortality (HR = 0.97). Lower systemic inflammation partially mediated the LC9-mortality relationship (SII: 5%, PIV: 6.3%). Higher LC9 scores are associated with reduced liver fibrosis and improved survival in MASLD, with systemic inflammation partially mediating the mortality association.
Alzheimer's disease (AD) is a multifactorial and progressive neurodegenerative disorder characterized by complex interactions among amyloid-β (Aβ) deposition, tau protein hyperphosphorylation, neuroinflammation, oxidative stress, metal dyshomeostasis, and impaired autophagy. Increasing evidence positions neuroinflammation not merely as a secondary response but as a central driver of disease progression, dynamically interacting with amyloid and tau protein pathology and contributing to synaptic dysfunction and neuronal loss. Among inflammatory mechanisms, microglial activation pathways-particularly TREM2 signaling, NLRP3 inflammasome activation, and complement cascade dysregulation-are currently the most clinically actionable targets, supported by genetic, biomarker, and therapeutic evidence. Emerging data suggest that modulation of innate immune pathways is most likely to confer benefit during the prodromal and early symptomatic stages of AD, when neuroinflammatory responses remain partially adaptive and neuronal networks retain functional reserve. Despite decades of drug development, many candidates have failed due to limited efficacy or safety concerns. Recent FDA approvals of anti-amyloid monoclonal antibodies, including aducanumab and lecanemab, represent important advances toward disease-modifying therapy, although their long-term clinical impact and safety profiles remain under evaluation. These developments underscore the importance of biomarker-guided patient selection, disease-stage stratification, and vigilant safety monitoring, particularly regarding amyloid-related imaging abnormalities. Therapeutic strategies are increasingly shifting toward multi-target approaches that integrate amyloid modulation, tau protein-directed interventions, and attenuation of maladaptive neuroinflammatory responses. Concurrently, inflammatory mediators and peripheral metabolic biomarkers are gaining recognition as tools for early detection, risk stratification, and therapeutic response monitoring, potentially enabling precision-based intervention. This review synthesizes current understanding of AD pathogenesis through an inflammation-centered framework, highlighting clinically actionable immune pathways and stage-specific therapeutic windows. By integrating mechanistic insights with biomarker-driven strategies, we aim to delineate translational paths toward more precise, safe, and clinically meaningful disease modification.
Medicinal plants have been used since ancient times for the treatment and prevention of various diseases. Plant-derived compounds offer a promising alternative to synthetic anti-inflammatory drugs, with potent activity and reduced toxicity. The seagrass Halodule uninervis exhibits several pharmacological properties due to the presence of various bioactive metabolites. However, its anti-inflammatory activity is not yet explored. In this study, we investigated the anti-inflammatory potential of an ethanolic extract of H. uninervis (HUE) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages by assessing its effect on the expression and activation of different inflammatory mediators. Our results revealed that HUE inhibited the LPS-stimulated inflammatory response in macrophages by downregulating the expression of iNOS and COX-2 and reduced the expression of NO and pro-inflammatory cytokines, IL-6 and TNF-α. Moreover, the anti-inflammatory potential of HUE was associated with reduced cellular migration and the suppression of the NF-κB, STAT3, and MAPKs pathways. Altogether, H. uninervis exhibits anti-inflammatory activity in vitro and may contribute to the modulate of inflammatory responses.
Diarrhea is characterized by excessive intestinal secretion and motility, often linked to inflammatory activation. Although synthetic antidiarrheal drugs are effective, their prolonged use can cause side effects, underscoring the need for safe, natural alternatives.This study investigated the antidiarrheal and anti-inflammatory activities of Bacillus subtilis DKU_09-fermented soybean (FS) powder in a mouse model of castor oil-induced diarrhea.Male ICR mice were orally administered FS (100, 200, or 300 mg/kg) for 7 days before the induction of diarrhea with castor oil. Diarrheal onset, frequency, fecal water content, and gastrointestinal (GI) transit were recorded. Inflammatory mediators in the small intestine were analyzed using real-time polymerase chain reaction and Western blotting, while serum immunoglobulin A (IgA) and immunoglobulin G (IgG) were measured by enzyme-linked immunosorbent assay.Pretreatment with FS significantly delayed the onset of diarrhea (FS200: 132.0 ± 9.8 min vs. castor oil-treated control [DC]: 76.9 ± 5.6 min, P < .05) and reduced diarrheal frequency (FS200: 32.9 ± 5.4% vs. DC: 51.9 ± 6.1%, P < .05). FS normalized the accelerated GI transit and markedly suppressed intestinal inflammation. The expression of Il-1β, Cox-2, mPGES-1, and NF-κB (p65) mRNA was significantly decreased by FS, accompanied by inhibition of mitogen-activated protein kinase (MAPK) phosphorylation (ERK, JNK, and p38). Serum IgG and IgA levels were unaffected, indicating that the effects were localized to intestinal tissue.Soybeans fermented by B. subtilis DKU_09 exert potent antidiarrheal effects through suppression of the NF-κB/Cox-2/PGE2 and MAPK signaling pathways, thereby reducing intestinal inflammation and motility. These findings suggest that the FS used in the present study may serve as a safe and natural functional food for the management of acute diarrhea.
Cecal ligation and puncture (CLP) is widely used to develop polymicrobial sepsis model in rodents, yet conventional CLP is not advantageous for evaluating long-term outcomes because most animals succumb without clinically aligned treatment. We therefore implemented source control (SC) combined with antibiotic therapy after CLP to enable post-acute observation and characterize pulmonary immune alterations after abdominal sepsis. Twenty-eight-week-old male C57BL/6J mice underwent CLP. After 6 h, the necrotic cecum was resected, followed by peritoneal lavage, and antibiotics were administered for three days (SC group). Sham mice underwent matched laparotomies without CLP, and age-matched naïve mice that did not undergo surgery or treatment served as controls. Survival, systemic inflammation, bacterial burden, and organ injury were assessed up to day 14 after CLP. We also evaluated pulmonary inflammation, lung immune cell composition, immunoglobulin profiles in plasma, lung homogenates, and bronchoalveolar lavage fluid (BALF), as well as airway epithelial injury. SC improved survival from 0% to approximately 90% through day 14. Plasma IL-6 and C-reactive protein levels declined after the acute phase, and little to no bacterial burden was detected in peritoneal lavage fluid, blood, or lung homogenates on day 14. Despite apparent systemic recovery, SC mice exhibited persistent pulmonary inflammation, with elevated levels of lung inflammatory mediators and sustained accumulation of neutrophils and monocytes. In contrast, CD4+ T cells, CD8+ T cells, and B cells were reduced in the lungs on day 14. IgM and IgG were elevated in plasma and lung homogenates but remained unchanged in BALF. Conversely, IgA levels were preserved in plasma and lung homogenates but selectively reduced in BALF. SC mice also showed reduced airway epithelial expression of E-cadherin, Occludin, and polymeric immunoglobulin receptor (pIgR), suggesting epithelial injury and impaired airway IgA transport in the post-acute phase. We developed a clinically aligned preclinical model of source-controlled abdominal sepsis that enables evaluation of post-acute pathophysiology. Despite apparent systemic recovery, sepsis survivor animals exhibited persistent pulmonary inflammation, adaptive immune cell reduction, airway epithelial injury, reduced pIgR expression, and selective BALF IgA loss. These findings suggest prolonged disruption of airway mucosal immunity after abdominal sepsis.
Previous study demonstrated that Hypnea cervicornis lectin (HCA) reduces inflammation and nociception via interaction with the lectin carbohydrate-binding site, modulating gene expression of the interleukins IL-1β, TNF-α and of iNOS. This study evaluated the effect of HCA in the rat model of arthritis induced by zymosan in the tibiotarsal joint and the involvement of macrophage-derived mediators. In vitro, macrophages were stimulated with zymosan before being incubated with HCA and the supernatant was injected into the joint. In vivo, HCA was administered by intravenous route after intra-articular injection of zymosan, fMLP, or macrophages supernatant. Hypernociception, edema and leukocyte influx were evaluated in the joints, and inflammatory mediators in the macrophage supernatant or periarticular tissue. In vitro, HCA (100 µg/ml) reduced NO2- and IL-1β in the supernatant of macrophages. In vivo, HCA (3 mg/kg) reduced articular hypernociception, edema and leukocyte influx elicited by zymosan, fMLP or by the supernatant of zymosan activated macrophages, as well as the zymosan-induced rolling and adhesion, and gene expression of iNOS and IL-1. HCA attenuated edema and leukocyte influx in the periarticular tissue, revealing preserved chondrocytes in the cartilage. In conclusion, HCA exerts anti-inflammatory effect in the arthritis induced by zymosan in rat tibio-tarsal joints, involving NO and IL-1 released by resident macrophages.
Ischemic stroke is an acute cerebrovascular disease that remains a major cause of death and long-term disability worldwide. In China and across East Asia, Lumbricus (earthworm) has long been used in traditional medicine for the management of stroke and related cerebrovascular disorders. As a major bioactive component of Lumbricus, lumbrokinase exhibits thrombolytic and anticoagulant activities. The present study investigated whether LK exerts neuroprotective effects in ischemic stroke by modulating TLR4/NF-κB-related inflammatory signaling. This study aimed to elucidate the neuroprotective mechanism of lumbrokinase against ischemic stroke. Candidate targets and signaling pathways were predicted by network pharmacology, and binding interactions were assessed by molecular docking. A rat middle cerebral artery occlusion (MCAO) model was then established to evaluate the neuroprotective efficacy of lumbrokinase. Neurological deficits, infarct volume, pathway-related proteins, and inflammatory cytokines were evaluated by behavioral tests, TTC staining, Western blot, and ELISA. Network analysis identified 49 overlapping inflammation-related targets significantly enriched in TLR4/NF-κB-related pathways. Molecular docking revealed favorable binding affinities (all < -5.0 kcal/mol) between lumbrokinase and core inflammatory mediators, including PTGS2, TRAF6, MyD88, and TLR4. In the MCAO model, lumbrokinase improved neurological function and reduced infarct volume across three doses, with the medium dose (12,000 U/kg) providing the most consistent benefit. Lumbrokinase also decreased the expression of TLR4, MyD88, TRAF6, TAK1 and p-NF-κB/NF-κB, as well as brain levels of TNF-α, IL-1β and IL-6. Lumbrokinase may confer neuroprotection in ischemic stroke, partly in association with attenuation of TLR4/NF-κB-related neuroinflammation in the ischemic brain.