NAD+

Abstract

Oxidative/anti-oxidative stress unbalance is one of the mechanisms of chronic obstructive pulmonary disease (COPD). Anion has been shown to be effective to eliminate reactive oxygen species, yet it is unknown if inhalation of anion (IA) can be beneficial for COPD intervention.

Abstract

Chronic liver disease (CLD) poses a significant global health challenge, and liver fibrosis is a crucial process in the pathogenesis of CLD. However, effective interventions to halt and reverse the progression of liver fibrosis remain elusive. This study investigated the potential of the nicotinamide phosphoribosyltransferase (NAMPT) inhibitor FK866 in treating diethylnitrosamine (DEN)-induced liver fibrosis in mice. We first demonstrated that DEN-induced hepatic fibrosis in mice was accompanied by upregulation of hepatic NAMPT and poly (ADP-ribose) polymerase 1 (PARP1) expression. Administration of FK866 inhibited the increase in alanine aminotransferase and aspartate aminotransferase levels and reversed the histopathological changes associated with DEN-induced liver fibrosis. It also suppressed the elevated expression of fibrotic markers, such as fibronectin, collagen IV, laminin, and α-smooth muscle actin. Further studies revealed that this therapeutic effect was achieved by inhibiting the NAD+ level, as well as the protein expression of NAMPT, PARP1, and inflammatory factors, including interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α, and P65. In conclusion, FK866 exhibits therapeutic potential for the treatment of liver fibrosis.

Abstract

Lactobacillus plantarum (L. plantarum) is a probiotic bacterium with diverse health-promoting effects. Recent evidence suggests that these benefits are mediated by extracellular vesicles (EVs) secreted by the bacterium; however, the underlying molecular mechanisms in the context of pathogenic inflammation remain poorly understood.

Abstract

Haemonchus contortus, a highly pathogenic nematode of small ruminants, causes substantial economic losses worldwide. The transition from the infective third-stage larva (L3) to the metabolically active fourth-stage larva (L4) is critical for parasite survival, as L4 larvae initiate blood-feeding within the host. This study aimed to optimize in vitro cultivation conditions for L4 larvae and evaluate their metabolic activity, providing a basis for future research on parasite biology and anthelmintic development. We systematically tested six culture media (BME, DMEM, IMDM, RPMI 1640, M199, LB broth; LB without FBS) supplemented with antibiotics including gentamicin (significantly improved L4 development: 85 % vs 70 % without, p < 0.05) to determine optimal L3-to-L4 conditions. Larval survival was assessed by motility under inverted microscopy; L4 development by exsheathment, body thickening (600-800 μm), and genital primordium visibility. To assess metabolic status, we probed proteolytic activity, which represents a key function of blood-feeding larvae. A fluorogenic peptide substrate was employed to detect significant cysteine protease activity, serving as a marker of a competent digestive system. These findings establish a refined methodology for in vitro cultivation of L4 larvae and highlight their suitability for functional studies and drug screening. By optimizing culture conditions and demonstrating active proteolysis, this study provides a valuable platform for investigating stage-specific parasite biology and identifying novel therapeutic targets.

Abstract

Atherosclerosis (AS), a chronic vascular pathology characterized by endothelial dysfunction, arises from the interplay of lipid dysregulation, oxidative stress, and inflammatory activation. Reactive oxygen species (ROS) overproduction triggers Nod‑like receptor protein 3 (NLRP3) inflammasome signaling, exacerbating inflammatory cascades that drive plaque progression. The nuclear factor erythroid 2‑related factor 2 (Nrf2)‑mediated antioxidant pathway serves as a critical counterbalance to ROS/NLRP3 axis dysregulation, positioning pharmacological Nrf2 activation as a promising therapeutic strategy. The present study investigated the anti‑atherosclerotic potential of ginkgolide C (GC), a terpene lactone from Ginkgo biloba with established anti‑inflammatory and anti‑ischemia/reperfusion injury properties, through coordinated modulation of redox‑inflammatory pathways. Complementary in vivo (high‑fat diet/vitamin D3‑treated ApoE-/- mice) and in vitro (oxidized‑low density lipoprotein‑stimulated aortic endothelial cells) models were established. Comprehensive analyses included histopathological characterization, lipid profiling, ultrastructural examination, redox‑inflammatory biomarker quantification, and molecular pathway validation. GC significantly attenuated hyperlipidemia and plaque progression while preserving vascular ultrastructure. Mechanistically, GC enhanced endothelial survival through dual pathway modulation: i) Nrf2 nuclear translocation upregulated antioxidant enzymes [heme oxygenase‑1/NAD(P)H quinone oxidoreductase 1/glutamate‑cysteine ligase modifier subunit], restoring redox homeostasis; ii) NLRP3 inflammasome inhibition via Caspase‑1 suppression mitigated inflammatory cytokine release. The present study demonstrated GC's dual‑target therapeutic efficacy against AS through Nrf2‑mediated oxidative stress resolution and NLRP3 inflammasome inactivation, offering new insights into phytochemical‑based cardiovascular interventions.

Abstract

Nicotinamide phosphoribosyltransferase (NAMPT) is the rate-limiting enzyme in the NAD+ salvage pathway and a promising therapeutic target in cancer. Resistance to NAMPT inhibitors, such as FK866, remains a key limitation to their clinical translation. While acquired resistance in cancer cell lines has been linked to target mutations, increased drug efflux, and metabolic reprogramming, innate resistance mechanisms have been poorly studied. Addressing this gap is crucial for identifying patient subgroups that are most likely to benefit from NAMPT-targeted therapies. Advanced castration resistance prostate cancer (CRPC) lacks effective targeted treatments. Among its heterogeneous subtypes, stem cell-like CRPC (CRPC-SCL) is characterized by independence from androgen receptor (AR) signaling, dependency on YAP/TAZ, and mesenchymal traits. In this study, we identify the YAP/nicotinamide N-methyltransferase (NNMT) axis as a key regulator of innate sensitivity to FK866 in stem-like mesenchymal CRPC cells. Using genetic and pharmacological models, we show that YAP or NNMT silencing rescues PC3 cells from FK866-induced apoptosis, endoplasmic reticulum stress, and NAD(H) depletion. Metabolomic profiling confirmed that NNMT activity depletes nicotinamide, sensitizing cells to FK866. We further validated NNMT upregulation across clinical CRPC-SCL datasets, where it strongly correlates with mesenchymal and therapy-resistant phenotypes. Murine prostate cancer cells with mesenchymal/stemness phenotypes (DVL3-SCM), that exhibit NNMT overexpression and high aggressiveness in vivo, also show increased sensitivity to FK866 compared with their parental counterparts (DVL3-PAR). In conclusion, we identify the YAP/NNMT axis as a determinant of innate sensitivity to NAMPT inhibition in prostate cancer. These findings support the use of NNMT as a predictive biomarker for NAD+-targeting therapies and provide mechanistic insight into a metabolic vulnerability of the CRPC-SCL subtype. Targeting the YAP/NNMT/NAMPT axis may represent a novel strategy for treating stem-like/mesenchymal, therapy-resistant prostate cancers.

Abstract

Glaucoma is characterized by progressive retinal ganglion cell degeneration. Nicotinamide supplementation has demonstrated neuroprotective potential in glaucoma by raising retinal and optic nerve nicotinamide adenine dinucleotide (NAD) via the salvage pathway, dependent on nicotinamide phosphoribosyltransferase (NAMPT). The NAMPT is essential for retinal function, and its extracellular form (eNAMPT) has been detected in blood. Reduced circulating eNAMPT could indicate impaired NAD biosynthetic capacity and glaucomatous neurodegeneration susceptibility.

Abstract

This review synthesizes research on nuclear factor erythroid 2-related factor 2 (Nrf2) in intestinal health across human, livestock, and mouse models. The Nrf2 signaling pathway serves as a master regulator of cellular antioxidant defenses and a key therapeutic target for intestinal inflammatory disorders, including ulcerative colitis and Crohn's disease. The interplay between oxidative stress, Nrf2 signaling, and NF-κB inflammatory cascades represents a critical axis in the pathogenesis and resolution of intestinal inflammation. Under normal physiological conditions, Nrf2 remains sequestered in the cytoplasm by Kelch-like ECH-associated protein 1 (Keap1), which facilitates its ubiquitination and proteasomal degradation. However, during oxidative stress, reactive oxygen species (ROS) and electrophilic compounds modify critical cysteine residues on Keap1, disrupting the Keap1-Nrf2 interaction and enabling Nrf2 nuclear translocation. Once in the nucleus, Nrf2 binds to antioxidant response elements (ARE) in the promoter regions of genes encoding phase II detoxifying enzymes and antioxidant proteins, including heme oxygenase-1 (HO-1), NAD(P)H quinone oxidoreductase 1 (NQO1), and glutamate-cysteine ligase. This comprehensive review synthesizes current evidence demonstrating that activation of Nrf2 signaling confers protection against intestinal inflammation through multiple interconnected mechanisms: suppression of NF-κB-mediated pro-inflammatory cascades, enhancement of cellular antioxidant capacity, restoration of intestinal barrier integrity, modulation of immune cell function, and favorable alteration of gut microbiota composition. We systematically examine a diverse array of therapeutic agents targeting Nrf2 signaling, including bioactive peptides, natural polyphenols, flavonoids, terpenoids, alkaloids, polysaccharides, probiotics, and synthetic compounds. The mechanistic insights and therapeutic evidence presented underscore the translational potential of Nrf2 pathway modulation as a multi-targeted strategy for managing intestinal inflammatory conditions and restoring mucosal homeostasis.

Abstract

Acrylamide (ACR), a common environmental and dietary neurotoxicant, exerts profound deleterious effects on the central nervous system by triggering oxidative stress, neuroinflammation, apoptosis, and motor impairments. Eugenol (EU), a natural phenolic compound known for its antioxidant and anti-inflammatory properties, was evaluated for its neuroprotective efficacy in ACR-induced brain toxicity in rats. Male Wistar rats were orally administered ACR (20 mg/kg/day) for four weeks to induce neurotoxicity, with concurrent administration of EU at two doses (50 and 100 mg/kg/day). Behavioral assessments, including foot splay, gait score, and rotarod performance, were conducted to evaluate motor coordination and neuromuscular integrity. Biochemical analyses revealed that ACR significantly elevated markers of oxidative and nitrosative stress, suppressed antioxidant defense mechanisms. Furthermore, ACR induced significant upregulation of pro-inflammatory mediators (NLRP3, p-NF-κB, IL-1β), as well as apoptosis markers such as caspase-3, alongside prominent histopathological alterations and astrocyte activation (evidenced by increased GFAP expression). Treatment with EU resulted in a dose-dependent amelioration of these neurotoxic effects. Notably, EU restored motor function, attenuated oxidative/nitrosative damage, and reactivated the Nrf2/NQO1/HO-1 antioxidant pathway. Simultaneously, it significantly downregulated the expression of NLRP3, p-NF-κB, and IL-1β, indicating strong anti-inflammatory action. Histological analysis confirmed preservation of neuronal architecture, while immunohistochemistry showed reduced caspase-3 and GFAP expression in EU-treated groups. These findings suggest that EU exerts potent neuroprotective effects against ACR-induced brain toxicity, primarily through modulation of redox balance, suppression of neuroinflammation, and inhibition of apoptotic cell death, via targeting both the Nrf2-mediated antioxidant system and the NLRP3/NF-κB/IL-1β inflammatory cascade.

Abstract

Glycopeptide antibiotics serve as critical last-resort drugs against resistant pathogens. However, emerging resistance─primarily due to altered interactions between glycopeptides and their bacterial targets─has begun to compromise their clinical effectiveness. Understanding the dynamic, molecular-level interactions between glycopeptide antibiotics and their targets is essential for elucidating resistance mechanisms and guiding the design of next-generation therapeutics. Conventional techniques, however, lack the resolution to capture these transient binding events in real time. In this study, we present a nanopore-based biomimetic probe strategy to study glycopeptide-peptidoglycan interactions at the single-molecule level. A series of biomimetic peptidoglycan precursor peptides (BPGPPs) with the sequence of (DE)nADEK(G5)DADA have been designed. Using these BPGPPs and the α-hemolysin (α-HL) nanopore, we achieved real-time single-molecule recordings of BPGPP-glycopeptide complex dissociation for a range of commercially available glycopeptides. The binding affinity of each glycopeptide toward BPGPPs is reflected by the mean dwell time of the complex within the α-HL nanopore. Furthermore, by investigating terminal residue mutations in BPGPPs, we gained insight into natural resistance mechanisms and potential trends in future glycopeptide-resistant strains. This research offers a time- and space-resolved single-molecule method for investigating glycopeptide-peptidoglycan interactions and holds promise for guiding the development of next-generation antimicrobial agents.

Abstract

Streptococcus pneumoniae (S. pneumoniae) remains a predominant cause of high morbidity and mortality in childhood and the elderly, despite the widespread pneumococcal conjugate vaccines (PCVs) vaccination through the world. The critical role of capsule in the pathogenicity of S. pneumoniae makes it an attractive drug target for alternative strategies to combat antibiotic-resistant and non-vaccine serotype infections. Here, we identified the natural compound molecule stevioside as an effective capsule inhibitor that reduces the biosynthesis of capsular polysaccharide through interfering with pyruvate metabolism and subsequent disruption of bacterial NAD + /NADH redox balance and energy generation. In vitro, the compound significantly sensitized streptococci to stress attacks and the killing of antibacterial peptides (AMPs). Meanwhile, capsule-mediated resistance to complement deposition, epithelial adherence and phagocytosis were all remarkably attenuated by stevioside. In vivo, stevioside treatment systematically protected mice from lethal streptococcal pneumoniae, as evident by an increased survival rate, alleviated pathological damage and inflammation level. Overall, the study provides stevioside as a promising lead compound for the further development of chemical capsule inhibitors aimed at curbing S. pneumoniae infections, and reveals a novel strategy for the discovery of S. pneumoniae capsule inhibitors based on pyruvate metabolism pathways.

Abstract

Sarcopenia is a degenerative skeletal muscle disorder closely associated with aging, characterized by the gradual loss of muscle mass and function, which severely impacts the quality of life in the elderly. In general, the severity of sarcopenia varies significantly among different patients and across different muscle groups, which increases the complexity and challenge of sarcopenia treatment. Based on the core pathological mechanism of sarcopenia, namely the deficiency of Nicotinamide adenine dinucleotide (NAD(+)) induced mitochondrial dysfunction, this study has developed a novel Energy-replenishing hydrogel microsphere (NMN@Lipo-s@AHM) for the targeted delivery of Nicotinamide Mononucleotide (NMN) to muscle cells through local injection. The stability of NMN was enhanced through liposomal encapsulation, peptide SS-31 was applied for mitochondrial targeting, and sustained local releasing was achieved via aldehyde hyaluronic acid methacrylate hydrogel microspheres (AHM). In vitro and in vivo experiments demonstrated that the Energy-replenishing hydrogel microspheres significantly alleviated dexamethasone-induced mitochondrial dysfunction and senescent phenotypes in muscle cells. Transcriptomic and proteomic analyses revealed that the hydrogel microsphere regulates mitochondrial function by activating the "AMPK-SIRT1-PGC1α" signaling pathway, thereby synergistically improving mitochondrial energy metabolism and cellular senescence. This study not only provides an efficient targeted delivery strategy for NAD+ supplementation but also offers new directions for the mechanistic research and clinical intervention of sarcopenia.

Abstract

Accumulation of superoxide radicals leads to disrupted redox signaling and oxidative damage. The primary extracellular scavenger of superoxide is extracellular superoxide dismutase (SOD3), a crucial enzyme in maintaining antioxidant status and proper immune function. SOD3 distribution to the extracellular matrix is determined by the presence of a C-terminal heparin-binding domain (HBD). This region can be removed through intracellular proteolytic processing by furin. Cleavage of the HBD has been shown to be modulated by post-translational cysteine redox status, regulating the secretion of SOD3. Interestingly, other members of the SOD family, SOD1 and SOD2, are known to be inhibited by lysine acetylation, a metabolically linked post-translational modification (PTM) that can alter protein structure, function, and localization. Yet, no reports describe the effect of acetylation on SOD3. Here, immunoblotting and mass spectrometry (MS) were used to quantify the global and site-specific acetylation of recombinant human SOD3. Interestingly, a predicted and targeted parallel reaction monitoring (PRM) MS-based approach was necessary to identify lysine acetylation within the C-terminal HBD of SOD3. Acetylation was found to prevent furin cleavage with no impact on SOD3 activity. Our results also reveal that SOD3 is robustly deacetylated by NAD+-dependent sirtuins (SIRT1 and SIRT3), with moderate activity against K220 and high activity against K211 and K212 in the HBD furin cleavage region. These sites of acetylation have not been previously reported, likely due to the peptide's unique hydrophilic nature. Overall, our findings reveal that sirtuin-directed deacetylation of SOD3 restored furin cleavage, defining an important link between redox homeostasis and acetylation-directed metabolic regulation of extracellular oxidative stress.

Abstract

Beige adipocytes are inducible thermogenic fat cells that emerge within white adipose tissue (WAT) in response to thermogenic stimuli and confer metabolic benefits. However, obesity impairs the generation of beige adipocytes, and the underlying mechanisms remain poorly understood. Here, we show that obesity leads to a loss of adipose progenitor cells (APCs) in WAT, accompanied by reduced estrogen (E2) levels and nicotinamide phosphoribosyltransferase (NAMPT) expression. Supplementation with E2 or nicotinamide mononucleotide (NMN), an NAMPT-derived nicotinamide adenine dinucleotide (NAD+) precursor, restores beige adipogenesis in diet-induced obese mice. Mechanistically, estrogen receptor α (ERα) in APCs is required for beige fat formation by promoting Nampt transcription. We further demonstrate that NAMPT is both necessary and sufficient to drive APC proliferation and differentiation, with interleukin-33 (IL-33) acting downstream to mediate these effects. These findings uncover a critical ERα/NAMPT/IL-33 axis that preserves progenitor function and thermogenic capacity, offering a potential therapeutic strategy to combat obesity-induced beige fat failure and associated metabolic dysfunction.

Abstract

Triple-negative breast cancer (TNBC) is characterized by aggressive behavior and high recurrence rates, contributing to poor prognoses of TNBC patients. However, the lack of appropriate molecular targets limits the effectiveness of current antineoplastic therapies. Therefore, effective therapeutic strategies are urgently needed. Melatonin (N-acetyl-5-methoxytryptamine) has shown a broad spectrum of anticancer activities, but its potential for treating TNBC remains elusive. In this study, we discovered that melatonin suppressed the growth and invasiveness of TNBC cells through downregulating glycolytic capacity in association with inhibition of Yes-associated protein 1 (YAP1) signaling. Notably, melatonin suppressed expression of nicotinamide phosphoribosyltransferase (NAMPT), an enzyme participated in nicotinamide adenine dinucleotide (NAD) turnover and contributes to protein poly(ADP)-ribosylation (PARylation). As a result, melatonin potentiated DNA damage and elevated apoptosis. Furthermore, it is found that melatonin suppressed NAMPT expression via inhibition of YAP1. Moreover, downregulation of glycolysis decreased protein PARylation levels and increased DNA damage accumulation, suggesting that melatonin suppresses the crosstalk between glycolysis and DNA repair signaling. Combined treatment with melatonin and Olaparib, an inhibitor of the major conductor of protein PARylation, poly(ADP-ribose) polymerase (PARP), showed additive inhibitory effects on breast cancer proliferation compared to their single treatment. These findings demonstrated that melatonin may be a promising agent for targeting YAP1-mediated glycolysis and DNA repair in breast cancer for enhancing the therapeutic efficacy of Olaparib in TNBC patients.

Abstract

Background and Objectives: Diabetic nephropathy (DN) is a major cause of end-stage renal disease, yet its molecular basis remains unclear. Nicotinamide adenine dinucleotide (NAD+) metabolism is crucial for energy regulation, redox balance, and inflammation. This study investigated the dysregulation of key NAD+ salvage enzymes (CD38, NAMPT, and SIRT1) across albuminuria stages in type 2 diabetes (T2D). Materials and Methods: A cross-sectional study was conducted on 225 participants: healthy controls (n = 45), T2D with normoalbuminuria (n = 60), microalbuminuria (n = 60), and macroalbuminuria (n = 60). Serum CD38, NAMPT, and SIRT1 were measured by ELISA, while CD38 and SIRT1 gene expression in peripheral blood mononuclear cells was analyzed by qPCR. Results: CD38 and NAMPT levels increased progressively with albuminuria, whereas SIRT1 levels declined significantly. CD38 and NAMPT correlated positively with HbA1c, creatinine, and urinary albumin-to-creatinine ratio (UACR), while SIRT1 showed inverse correlations and a positive association with eGFR. Regression analysis identified CD38 and NAMPT as independent positive predictors of albuminuria, and SIRT1 as a negative predictor. ROC analysis revealed strong diagnostic performance for CD38 (AUC = 0.89) and SIRT1 (AUC = 0.88). Conclusions: These findings highlight disrupted NAD+ salvage pathways in DN and suggest that restoring NAD+ balance, through CD38 inhibition, SIRT1 activation, or NAD+ precursor supplementation, may offer promising renoprotective strategies.

Abstract

Hepatocellular carcinoma (HCC) is frequently linked to compensatory proliferating hepatocytes in damaged livers, yet the underlying molecular mechanisms remain elusive. The Mediator complex precisely coordinates multiple transcription factors and cofactors to regulate diverse physiological and pathological processes. Here, we discovered that Mediator subunit MED23 is involved in the progression of HCC. Both constitutive and inducible liver-specific ablation of Med23 effectively inhibited HCC development in diethylnitrosamine (DEN)-induced HCC mouse models. Mechanistically, MED23 deficiency significantly compromised hepatocyte cell viability by reducing the stability of the NQO1 protein, thereby leading to an increase in reactive oxygen species (ROS) production. Furthermore, MED23 collaborates with the transcription factor RFX5 to regulate a novel enhancer function for IGF2 expression, which thus influences hepatocyte viability and HCC development. Consistently, overexpression of IGF2 in MED23-deficient HCC cells stabilizes NQO1 and partially restores cell growth and reduces apoptosis. Collectively, our findings underscore the significance of the MED23-IGF2-NQO1 axis in HCC progression and propose a novel therapeutic strategy for the treatment of HCC.

Abstract

The combination of PARP and NAMPT inhibitors (PARPi/NAMPTi) has been explored for the treatment of triple-negative breast cancer, Ewing sarcoma and high-grade serous carcinoma (HGSC). However, dose limiting toxicity has hampered NAMPTi in clinical trials. To maximise the therapeutic window, we set out to identify predictive genomic biomarkers. Bioinformatic analysis and screening of a panel of epithelial ovarian cancer (EOC) cell lines revealed that cells with RAS/PI3K pathway mutations are sensitive to the NAMPTi FK866. Combined exposure to olaparib and FK866 is associated with a reduction in nicotinamide mononucleotide (NMN) and the PARP substrate nicotinamide adenine dinucleotide (NAD+), with coincident increases in ROS production, DNA damage and apoptosis induction. Caspase 3/7 activity is upregulated to a greater extent in RAS/PI3K mutant cell lines. Finally, the combination significantly reduces omental tumour weight and increases overall survival in mice injected with ID8 Trp53-/-;Pten-/- cells. This study highlights the potential of the PARPi/NAMPTi combination in RAS/PI3K pathway mutant EOC.

Abstract

Alzheimer's disease (AD) poses significant challenges to public health and well-being, with current treatments often providing limited efficacy. Chuanzhitongluo capsule (CZTL) has neuroprotective effects, and is expected to be used for the treatment of AD. In this study, the chemical composition, pharmacological effects and underlying mechanisms of CZTL against AD were systematically investigated. We identified the major components of CZTL through ultra-high-performance liquid chromatography coupled with a quadrupole time-of-flight mass spectrometer. AD rat model was established via scopolamine injection, followed by the administration of CZTL at various dosages, and pharmacological effects were then systematically evaluated. Furthermore, the potential mechanisms were explored using metabolomics and immunohistochemistry. Seventeen chemical constituents were identified from CZTL. Pretreatment with CZTL led to significant improvements in cognitive function and reductions in neuronal loss among AD rats. CZTL administration decreased abnormal protein aggregates (Aβ and Tau), along with markers of oxidative stress and inflammation. Metabolomic and immunohistochemical analyses indicated that CZTL modulated nicotinamide metabolism and impacted levels of NAD+, UMP, nitric oxide, and SIRT1 activity. These results suggest that CZTL effectively mitigates cognitive deficits and neuronal loss in AD by regulating nicotinamide and SIRT1, while inhibiting oxidative stress and inflammation. This study lies in their contribution to the development of novel anti-AD therapies derived from traditional Chinese medicine, paving the way for new approaches in managing neurodegenerative diseases.

Abstract

Acute myeloid leukemia (AML) cells depend on nicotinamide adenine dinucleotide (NAD+) biosynthesis via nicotinamide phosphoribosyltransferase (NAMPT) for survival. Single-cell RNA sequencing revealed robust NAMPT expression across diverse AML subtypes. Proteomic profiling showed that NAMPT inhibition with KPT-9274 induced adaptive upregulation of BCL2, an anti-apoptotic protein, highlighting a survival mechanism. BH3 profiling confirmed that AML cells hierarchically depend on BCL2, followed by MCL1 and BCLxL, for survival. Combining KPT9274 with the BCL2 inhibitor venetoclax synergistically enhanced mitochondrial dysfunction, cytochrome C release, and apoptotic death in AML blasts. Additionally, NAMPT inhibition reduced PARP activity and impaired DNA repair pathways, sensitizing AML cells to cytarabine and hypomethylating agents. Together, these results demonstrate that NAMPT inhibition both potentiates venetoclax activity and enhances the cytotoxic effects of standard chemotherapies by targeting metabolic and DNA repair vulnerabilities. These findings provide strong preclinical support for evaluating NAMPT and BCL2 dual inhibition strategies in future AML clinical trials.

Abstract

Ionizing radiation (IR) is well documented for its gonadotoxic effects, resulting in ovarian dysfunction and infertility through direct cellular damage and inflammatory processes. The activation and regulation of the NOD-like receptor protein 3 (NLRP3) inflammasome are crucial for modulating pyroptosis and inflammatory responses. Recent findings suggest that acetylation is essential for the full activation of the NLRP3 inflammasome. This study examines the protective effects of Nicotinamide mononucleotide (NMN) against IR-induced pyroptosis in ovarian granulosa cells, with a particular focus on the molecular mechanisms involving NLRP3 protein acetylation. Our observations from both in vivo and in vitro experiments demonstrate that IR exposure results in increased pyroptosis of granulosa cells, evidenced by elevated levels of NLRP3, Caspase-1, IL-1β, and IL-18, alongside a reduction in ovarian reserve. Importantly, SIRT3, an NAD+-dependent deacetylase, plays a role in mitigating the decline in ovarian function by promoting the deacetylation of NLRP3 at lysine 570, thereby inhibiting the activation of the NLRP3 inflammasome. This study provides valuable insights into the potential of NMN supplementation in mitigating the adverse effects of radiation on ovarian function.

Abstract

1-Bromopropane, an alternative to ozone-depleting solvents, exhibits neurotoxicity in humans and rats. The aim of the present study was to identify the genes or signaling pathways involved in the neurotoxicity and hepatotoxicity of 1-bomopropane in two inbred strains of rats. F344 rats and WNA/NUM rats were exposed to 1-bromopropane or filtered air by inhalation for either a single 8-hour session, or 8 h daily for 4 weeks. Motor nerve conduction velocity and distal latency were measured in the tail. At the end of the experiment, the animals were decapitated, and the hippocampus, liver, and blood were collected. Exposure to 1-bromopropane for 4 weeks increased distal latency in the tail nerve significantly in F344 and marginally in WNA/NUM. It also increased total and direct bilirubin in both rat strains. Eight-hour exposure increased metallothionein 2A, metallothionein 1 and NAD(P)H quinone dehydrogenase 1 expression in the liver of both rat strains, whereas 4-week exposure upregulated glutathione S-transferase alpha 3 and CD36 molecule in the liver of both strains. KEGG analysis showed that 8-hr 1-bromopropane upregulated pathways of apoptosis, NOD-like receptor signaling and colorectal cancer, in both the hippocampus and liver, whereas 4-week exposure upregulated NOD-like receptor signaling pathway both in the hippocampus and liver of the two rat strains. Insulin signaling pathway was upregulated persistently in the hippocampus of the two rat strains. Our results suggest the involvement of immune system/inflammation-related pathway in the neuro- and hepato-toxicity of 1-bromopropane and the involvement of insulin signaling pathway in the neurotoxicity of 1-brromopropane.

Abstract

Sarcopenia contributes to all-cause mortality in the elderly; however, there is no specific treatment. Mesenchymal stromal cells (MSCs) ameliorate age-related muscle loss and dysfunction and are potential therapeutic candidates for sarcopenia. However, their activity is easily affected by the surrounding environment and they are prone to replicative senescence during in vitro culture. Therefore, a drug that delays aging and enhances its function is required. Here, we investigated whether nicotinamide adenine dinucleotide (NAD+) pretreatment enhances the therapeutic efficacy of MSCs on skeletal muscle atrophy and its underlying mechanism in a D-galactose (D-gal)-induced mouse model.

Abstract

Nicotinamide mononucleotide (NMN), a potent nicotinamide adenine dinucleotide (NAD+) precursor, has demonstrated significant potential in mitigating mitochondrial oxidative stress, alleviating degenerative diseases, and reducing reactive oxygen species (ROS) generation. However, its topical application is hindered by inherent challenges, including high hydrophilicity, poor skin permeability, and limited stability. To address these limitations, we developed elastic cationic liposomal nanogels (EC-Lip-nanogels), a novel delivery system composed of Nap-FF-GHK nanogels and elastic cationic liposomes (EC-Lips). Nap-FF-GHK peptide was synthesized by coupling the self-assembled scaffold Nap-FF with the anti-aging module GHK, which would be self-assembled into cationic nanogels. EC-Lips was constructed by incorporating edge activators (EAs) into cationic liposomes (C-Lips). The NMN-loaded EC-Lip-nanogels were systematically characterized, demonstrating favorable physicochemical properties with an average particle size of 53.2 nm, a zeta potential of +14.0 mV, and a high encapsulation efficiency of 90.2%. Notably, the penetration enhancement achieved by EC-Lips and EC-Lip-nanogels was 22.7% and 16.0%, respectively, compared to free NMN. This study might establish EC-Lip-nanogels as a promising platform for improving the skin permeability of NMN and other hydrophilic bioactive molecules, offering a transformative approach for topical applications.

Abstract

Cerebral ischemia-reperfusion injury (CIRI) represents a critical barrier to effective stroke treatment, characterized by mitochondrial dysfunction and neuroinflammatory cascades. Although idebenone (IDE), a CoQ10 analog with potent antioxidant properties, has shown therapeutic potential, its poor mitochondrial accumulation and pharmacokinetic limitations have hindered clinical efficacy. To overcome these challenges, we synthesized W1a, a novel IDE derivative conjugated to the mitochondria-targeting peptide SS-20, designed to enhance mitochondrial localization and pharmacodynamic activity. We demonstrated that W1a significantly ameliorated glutamate-induced oxidative injury in HT22 hippocampal neurons by preserving mitochondrial membrane potential (MMP), enhancing adenosine triphosphate (ATP) production, reducing reactive oxygen species (ROS) accumulation, and maintaining mitochondrial morphology. In vivo, W1a conferred robust neuroprotection in a murine model of transient middle cerebral artery occlusion (tMCAO), as evidenced by reduced infarct volume, improved survival rates, and enhanced neurological recovery. Mechanistically, transcriptomic and biochemical analyses revealed that W1a restored mitochondrial gene expression, elevated NAD+/NADH and ATP levels, suppressed ROS overproduction, and downregulated pro-inflammatory signaling pathways. Moreover, W1a inhibited microglial and astrocytic activation, further supporting its anti-inflammatory efficacy. Collectively, our findings establish W1a as a potent neuroprotective agent that targets mitochondrial dysfunction and neuroinflammation drivers of CIRI pathology, offering a promising therapeutic strategy for ischemic stroke.

Abstract

SARM1 is a neuronal Nicotinamide adenine dinucleotide (NAD+) hydrolase that drives axonal degeneration and neuronal death by depleting NAD+, yet how NAD+ loss triggers axon loss and cell death has remained unclear. Here, we define a nonapoptotic death program downstream of endogenous SARM1 activation and NAD+ loss using a genetically tractable nonneuronal eHAP cell model. Upon NAD+ depletion, BAX is activated but caspase activation is suppressed due to APAF1 degradation via the E3 ligase HERC4, effectively uncoupling mitochondrial outer membrane permeabilization from apoptosome formation. Mechanistically, NAD+ depletion inhibits mTOR/AKT signaling, destabilizing MCL1 and relieving BAX from repression. We further identified Neurofibromatosis type II, NF2, as a regulator that promotes SARM1 transcription through the Hippo-YAP/TAZ pathway. The SARM1-dependent BAX activation and the role of NF2 in axon degradation were validated in neuronal models of axon degeneration. Together, these findings reveal how SARM1-driven metabolic collapse rewires cell death execution, positioning BAX, MCL1, APAF1, NF2, and HERC4 as core effectors in a nonapoptotic degenerative pathway linking metabolic stress to neurodegeneration.

Abstract

Colorectal cancer (CRC) poses a significant global health challenge, yet immune checkpoint blockade (ICB) therapy benefits only a small subset of patients with mismatch repair-deficient (dMMR) or microsatellite instability-high (MSI-H) tumours. Through analyses of public single-cell and spatial transcriptomic datasets, primary mouse cell sorting and adoptive transfer experiments, flow cytometry, multiplex immunofluorescence, immunohistochemistry, and coimmunoprecipitation, we revealed that sentrin-specific protease 7 (SENP7) promotes regulatory B-cell (Breg) differentiation and inhibits senescence by activating the expression of the NAD-dependent protein deacetylase sirtuin-1 (SIRT1) via deSUMOylation, thereby enhancing the expression of genes such as interleukin-10 (IL-10). Notably, targeting SENP7 in B cells improved the antitumour efficacy of anti-PD-1 therapy. These findings suggest that inhibiting SENP7 may offer a promising strategy to sensitize immunologically "cold" tumours to immune checkpoint blockade.

Abstract

Objective: To investigate the changes in hepatic phase II detoxification enzymes and their mechanism in metabolic associated steatohepatitis (MASH) induced by a methionine-choline-deficient (MCD) diet in mice. Methods: Ten C57BL/6J mice were randomly divided into two groups, with five mice in each group, and fed with a control diet (NCD group) and a methionine-choline-deficient diet (MCD group) for four consecutive weeks to establish the MASH model in mice. Mice body weight was recorded weekly. Mice peripheral blood and liver tissue samples were collected after four weeks. The liver histopathological changes were observed by hematoxylin-eosin staining and Sirius red staining in liver tissue. The levels of plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST) and triglycerides were measured by an automatic biochemical analyzer. Triglyceride and total cholesterol were used to evaluate the lipid accumulation condition in the liver of mice with Oil red O staining. Real-time fluorescence quantitative PCR was used to detect the expression of liver inflammatory factors interleukin (IL)-1β and monocyte chemoattractant protein-1 (MCP-1) condition. Transcriptome sequencing and bioinformatics were used to analyze the changes in gene expression profiles in the liver of mice and screen differentially expressed genes. The expression conditions of phase Ⅱ detoxification enzymes glutathione S-transferase mu 4 (GSTM4), dihydronicotinamide riboside:quinone oxidoreductases (NQO-2), sulfotransferase 1β1 (SULT1β1), and uridine diphosphate glucuronosyltransferase 2 family, polypeptide A3(UGT2A3) were verified by real-time fluorescent quantitative PCR. Plasma malondialdehyde content, total antioxidant capacity (T-AOC), plasma and liver glutathione content were determined using commercial kits. The expression of nuclear factor E2-related factor 2 (Nrf2), GSTM4, and UGT1A6 was examined by Western blotting. The independent sample t-test was used for comparison between the groups. Results: The body weight of mice in the MCD group showed a gradual downward trend, while the body weight of mice in the NCD group did not change significantly following four weeks of different dietary feeding. The MCD group mice liver had yellow-white appearance with round edges. The liver/body mass index was significantly lower in the NCD group (t=3.216, P<0.01). Hematoxylin-eosin staining showed that hepatocytes in the MCD group had an occurrence of fatty degeneration accompanied by inflammatory cell infiltration, with a higher NAFLD activity score (NAS) compared to the NCD group (t=7.155, P<0.001). Sirius red staining showed that the the liver of the MCD group had mildly increased periportal fibers. Plasma biochemical tests indicated that plasma ALT, AST, and triglyceride levels were significantly higher in the MCD group than those in the NCD group (t=8.920, P<0.001; t=6.696, P<0.001; t=3.904, P<0.01). Oil red O staining showed that a large number of lipid droplets accumulated in the liver tissue of the MCD group and were more severe than those in the NCD group (t=7.405, P<0.001). The triglyceride content was significantly higher in the liver of the mice in the MCD group than that in the NCD group (t=3.559, P<0.01), and the expression of inflammatory factors IL-1β and MCP-1 was significantly increased (t=2.562 and 2.391, respectively, P<0.05). Transcriptome sequencing analysis showed that the expression profile of genes related to lipid metabolism was changed in the liver tissue of the mice in the MCD group. The expression of multiple phase Ⅱ detoxification enzymes was significantly downregulated. Real-time fluorescence quantitative PCR verification demonstrated that the expression of four phase Ⅱ detoxification enzymes GSTM4, NQO2, SUIL1β1, and UGT2A3 were significantly lower in the liver of the mice in the MCD group than those in the NCD group (t=2.498, 3.570, 3.768, and 4.166, respectively, P<0.05). The detection kit showed that compared with the NCD group, the malondialdehyde content in the liver of mice in the MCD group increased (t=3.601, P<0.01), while the plasma total glutathione (t=11.93, P<0.001) and reduced glutathione levels were significantly reduced (t=3.635, P<0.01). The total antioxidant capacity of the liver decreased (t=2.872, P<0.05), and the total glutathione and reduced glutathione levels in the liver were significantly increased (t=3.175 and 3.064, P<0.05). Western blotting showed that the expression of Nrf2, GSTM4, and UGT1A6 proteins was significantly lower in the MCD group than that in the NCD group (t=3.385, 2.990, 2.168, P<0.05). Conclusions: The expressions of multiple phase Ⅱ detoxification enzymes and antioxidant capacity are reduced in the liver of MASH mice induced by the MCD diet, and its mechanism is related to the down-regulation of the expression of the upstream regulatory factor Nrf2 protein.

Abstract

The aim of this study was to explore the role and mechanism of SMP30 in diabetic cataract(DC) patients with different glycemic control levels and high glucose-stimulated human lens epithelial cells. First, aqueous humor and lens anterior capsules were collected during cataract surgery. Second, human lens epithelial B3(HLE-B3) cells were stimulated with different glucose concentrations. Then, SMP30-overexpressing plasmids were transfected into HLE-B3 cells. Measured blood and aqueous humor glucose concentrations in cataract patients. Oxidative stress levels was assessed by analyzing SOD, MDA and ROS. The mRNA and protein expression levels of SMP30 and Keap1/Nrf2/NQO1 pathway factors were detected. The cell viability was analyzed. The glucose concentration in blood and aqueous humor increased, and MDA content increased and SOD activity decreased in DC patients with poor glycemia control. However, SMP30 expression was upregulated of these patients, in parallel with the possible activation of Keap1/Nrf2/NQO1 pathway. The MDA content and ROS levels increased, and SOD activity decreased under high glucose(40 mM, 60 mM) in HLE-B3 cells. Meanwhile, SMP30 expression was up-regulated and Keap1/Nrf2/NQO1 pathway was activated at a glucose of 20 mM, but down-regulated and Keap1/Nrf2/NQO1 pathway was inhibited at a glucose of 40 mM or 60 mM. Overexpression of SMP30 reduced the oxidative stress damage and increased the cell viability, and reversed the expression of Keap1/Nrf2/NQO1 pathway factors induced by high glucose in HLE-B3. This study demonstrates that SMP30 may protect human lens epithelial cells from high glucose-stimulated oxidative stress damage and may involves the regulation of Keap1/Nrf2/NQO1 pathway.

Abstract

Previous studies have demonstrated that non-Saccharomyces yeasts can enhance malolactic fermentation (MLF) efficiency during co-fermentation. However, the underlying mechanisms remain unclear. Here, we demonstrate how specific strains create favorable metabolic niches through tripartite fermentation assays involving 16 strains from 8 species co-cultured with Saccharomyces cerevisiae F5 and Oenococcus oeni SD-2a, Hanseniaspora spp. emerged as optimal partners, reducing MLF duration by 50 % (≤48 h vs. 96 h in controls) while increasing O. oeni biomass by > 10-fold. Metabolomic analysis revealed that O. oeni utilizes malate as a carbon source within an ecological niche shaped by yeast-driven sugar competition, while upregulated nicotinamide metabolism enhanced NAD+ regeneration, boosting malolactic enzyme activity. Furthermore, strain-specific peptide secretion provided targeted bacterial support: H. osmophila NX39 produced quorum-sensing peptides and fumarate to activate bacterial pathways, whereas H. uvarum HN2 synthesized nutritional peptides and arabitol to alleviate auxotrophy. Exogenous amino acids (Glu/Trp/Cys) further enhanced MLF efficiency, with glutamate specifically accelerating early O. oeni growth while maintaining S. cerevisiae viability (stable at 108 CFU/mL). These findings transform co-inoculation from a high-risk practice into a robust enological strategy by establishing triple-strain simultaneous alcoholic-malolactic fermentation systems, providing a foundation for next-generation precision enology.