Introduction/Objectives Procyanidins are abundant dietary flavonoids with diverse pharmacological activities; however, their Absorption, Distribution, Metabolism, and Excretion (ADME) remain incompletely characterized. MT-8, a procyanidin B3 derivative with an ethyl group at the C8 position of the A-ring, is a promising preclinical candidate for cerebral ischemia treatment and is currently in the Investigational New Drug (IND) application stage. This study aimed to systematically characterize the ADME profile of MT-8 in male Sprague-Dawley (SD) rats following intravenous administration to provide a reference for ADME studies of procyanidin-related compounds. Methods A single intravenous dose of 24 mg/kg (100 mu Ci/kg) [C-14]MT-8 was administered to male SD rats. Given the observed instability of MT-8 in plasma due to catechol oxidation, 10 mM tris(2-carboxyethyl) phosphine hydrochloride (TCEP) was employed as a stabilizer in all sample preparations. Pharmacokinetics, tissue distribution, mass balance, metabolite profiling, and metabolic enzyme phenotyping were comprehensively evaluated. Results Pharmacokinetic analysis revealed rapid elimination of [C-14]MT-8-related substances, with a terminal half-life of 1.77 hours. The blood-to-plasma radioactivity ratio (1.68) indicated preferential distribution into blood cells. Tissue distribution revealed the highest radioactivity concentrations in the small intestine, kidneys, and liver. Mass balance analysis showed total radioactive recovery of 95.49% within 168 hours, with clearance of >90% of drug-related substances within 24 hours, predominantly via feces (85.57%) and bile (62.78% in bile duct-cannulated rats), with only 9.91% in urine. Metabolite profiling identified 32 metabolites in plasma, urine, feces, and bile, with methylation and glucuronidation as the major metabolic pathways. In vitro phenotyping referred to catechol-O-methyltransferase (COMT) involvement in methylation and UDP-glucuronosyltransferase 1A1 (UGT1A1) and 1A9 (UGT1A9) in glucuronidation. Discussion After verifying the mechanism of the instability of MT-8 in plasma, we tracked the in vivo disposition of MT-8 via radioisotope labeling technique and acquired credible pharmacokinetic, mass balance, tissue distribution, and metabolite identification results. Phenotypes of two enzymes were determined to further the investigation of the methylation and glucuronidation process of in vivo clearance of MT-8. Conclusion Collectively, MT-8 undergoes rapid clearance, broad distribution, extensive metabolism, and predominant biliary/fecal excretion. These findings provide critical support for MT-8's IND application and subsequent clinical development.

Vascular dementia (VaD) is the second most common type of dementia lacking effective treatments. Inulin-type hexasaccharide (IHS) extracted from traditional Chinese herbal medicine BaJiTian (Morinda officinalis) has antidepressant effects in many animal models. In this study, we aimed to investigate the therapeutic effects and potential mechanisms of IHS in VaD. Our results showed that IHS could effectively improve cognitive impairment in VaD mice, alleviate histological changes, and promote cerebral blood flow recovery. In addition, we observed that IHS could reduce neuroinflammation and apoptosis in VaD mice. The comprehensive characterization analysis of the serum, fecal, and brain tissue metabolites revealed that IHS regulates multiple metabolic pathways including sphingolipid metabolism, glycerophospholipid metabolism, and pyruvate metabolism in VaD mice. Furthermore, integrative analysis of metabolomics and brain proteomics found that IHS modulates the sphingolipid metabolism pathway to decrease accumulation of ceramide in the brains of VaD mice. Based on the role of ceramide in VaD development and cognitive impairment, IHS improves the pathology of VaD mice at least partially through the sphingolipid metabolism pathway. These results provide a framework for better understanding the mechanisms of the therapeutic effects of IHS in VaD.

Cardiac fibrosis remains a critical determinant of adverse outcomes in heart disease, yet effective anti-fibrotic therapies are lacking. While multiple semaphorin family members participate in cardiovascular pathophysiology, the role of semaphorin 4f (Sema4f) in cardiac fibrosis remains unexplored. This study investigates the role and mechanisms of Sema4f in fibrotic remodeling post-myocardial infarction (MI). We employed flow cytometry to characterize cell type-specific Sema4f expression patterns in post-MI hearts. Lineage-specific knockout mice (fibroblast vs. myeloid) were subjected to left anterior descending ligation to assess functional consequences. Proteomic analysis of Sema4f-deficient cardiac fibroblasts was conducted to identify downstream effectors. Key pathways were subsequently validated using pharmacological inhibitors. We found that Sema4f expression was markedly upregulated during the fibrotic phase post-MI, primarily due to fibroblast activation. Fibroblast-, but not myeloid-, specific Sema4f deletion significantly reduced fibrosis and improved cardiac function. Proteomic profiling revealed that Sema4f deficiency led to downregulation of pro-fibrotic gene expression, which was associated with impaired actin cytoskeletal remodeling and decreased nuclear translocation of YAP/TAZ. Pharmacological inhibition of either actin remodeling or YAP/TAZ activity attenuated fibrosis, whereas YAP/TAZ activation abolished the anti-fibrotic effects of Sema4f knockout. Our study provides the first evidence demonstrating the functional role of Sema4f in cardiac fibroblast activation and fibrosis progression. We have identified a fibroblast-specific mechanism mediated by the Sema4f-actin cytoskeleton-YAP/TAZ axis, offering novel mechanistic insights into fibrosis regulation and revealing a promising therapeutic target for cardiac fibrosis with potential clinical applications.

INTRODUCTION/OBJECTIVES:Procyanidins are abundant dietary flavonoids with diverse pharmacological activities; however, their Absorption, Distribution, Metabolism, and Excretion (ADME) remain incompletely characterized. MT-8, a procyanidin B3 derivative with an ethyl group at the C8 position of the A-ring, is a promising preclinical candidate for cerebral ischemia treatment and is currently in the Investigational New Drug (IND) application stage. This study aimed to systematically characterize the ADME profile of MT-8 in male Sprague-Dawley (SD) rats following intravenous administration to provide a reference for ADME studies of procyanidin-related compounds. METHODS:A single intravenous dose of 24 mg/kg (100 μCi/kg) [¹⁴C]MT-8 was administered to male SD rats. Given the observed instability of MT-8 in plasma due to catechol oxidation, 10 mM tris(2-carboxyethyl) phosphine hydrochloride (TCEP) was employed as a stabilizer in all sample preparations. Pharmacokinetics, tissue distribution, mass balance, metabolite profiling, and metabolic enzyme phenotyping were comprehensively evaluated. RESULTS:Pharmacokinetic analysis revealed rapid elimination of [¹⁴C]MT-8-related substances, with a terminal half-life of 1.77 hours. The blood-to-plasma radioactivity ratio (1.68) indicated preferential distribution into blood cells. Tissue distribution revealed the highest radioactivity concentrations in the small intestine, kidneys, and liver. Mass balance analysis showed total radioactive recovery of 95.49% within 168 hours, with clearance of >90% of drug-related substances within 24 hours, predominantly via feces (85.57%) and bile (62.78% in bile duct-cannulated rats), with only 9.91% in urine. Metabolite profiling identified 32 metabolites in plasma, urine, feces, and bile, with methylation and glucuronidation as the major metabolic pathways. In vitro phenotyping referred to catechol-O-methyltransferase (COMT) involvement in methylation and UDP-glucuronosyltransferase 1A1 (UGT1A1) and 1A9 (UGT1A9) in glucuronidation. DISCUSSION:After verifying the mechanism of the instability of MT-8 in plasma, we tracked the in vivo disposition of MT-8 via radioisotope labeling technique and acquired credible pharmacokinetic, mass balance, tissue distribution, and metabolite identification results. Phenotypes of two enzymes were determined to further the investigation of the methylation and glucuronidation process of in vivo clearance of MT-8. CONCLUSION:Collectively, MT-8 undergoes rapid clearance, broad distribution, extensive metabolism, and predominant biliary/fecal excretion. These finding.

Ligstroside (LIG) is a phenolic secoiridoid glycoside naturally occurring in olives and structurally related to the widely studied olive constituent oleuropein. Although LIG has been reported to exhibit diverse biological activities, it has received comparatively limited attention, and the molecular targets underlying its bioactivity remain largely unknown, thereby limiting exploration of its pharmacological potential. To facilitate target identification and mechanistic investigation, we designed and synthesized a LIG-derived photoaffinity probe (PAL-LIG) through minimal modification of the LIG scaffold. The probe incorporates a photoactivatable diazirine group that enables UVinduced covalent crosslinking with proximal proteins together with an alkyne handle that allows subsequent bioorthogonal conjugation via copper-catalyzed azide–alkyne cycloaddition (CuAAC). Following photo-crosslinking, probe-labeled proteins can be conjugated with azide-functionalized reporter tags, including biotin for streptavidinbased enrichment and pulldown or TAMRA for fluorescence detection and cellular imaging. Using this strategy, PAL-LIG enables enrichment of potential LIG-binding proteins as well as fluorescence-based visualization of probe labeling in cells. Importantly, PAL-LIG retains biological activity comparable to LIG, as demonstrated by its similar ability to promote ATP production in mouse C2C12 myoblasts, indicating that installation of the photoaffinity and alkyne handles minimally perturbs the intrinsic bioactivity of LIG. Collectively, PAL-LIG provides a chemical tool for investigating the direct protein targets and molecular mechanisms of LIG.

Obesity, a major global health challenge associated with metabolic and cardiovascular disorders, has drawn increasing attention to the therapeutic potential of white adipose tissue (WAT) browning. Although the lysine methyltransferase SETD7 has been implicated in various cardiovascular and metabolic diseases, its role in adipose thermogenesis remains unclear. Here, we reported that SETD7 was upregulated in inguinal WAT (iWAT) of obese mice and was primarily localized to mature adipocytes. Setd7 knockdown (Setd7⁺/⁻) mice exhibited enhanced thermogenic gene expression and iWAT browning upon cold exposure or β3-adrenergic stimulation, whereas thermogenic activity in brown adipose tissue (BAT) remained largely unaffected. In vitro, SETD7 knockdown did not alter adipogenesis but potently augmented thermogenic capacity in beige adipocytes, while SETD7 overexpression exerted the opposite effect. Mechanistically, RNA-Seq analysis revealed that SETD7 deficiency upregulated Adcy7 transcription, leading to increased Sirt1 levels and enhanced Creb1 phosphorylation, thereby activating the thermogenic program. Notably, Setd7⁺/- mice resisted high-fat diet (HFD)-induced obesity, exhibiting reduced weight gain, elevated energy expenditure, and improved metabolic health. Together, these findings identify SETD7 as a negative regulator of iWAT thermogenesis and suggest that targeting SETD7 may represent a promising strategy for combating obesity.

BackgroundEpigenetic regulation plays a pivotal role in adipocyte development and thermogenesis. Ash2l, a key component of the COMPASS (Complex of Proteins Associated with Set1) histone methyltransferase, regulates gene expression through epigenetic mechanisms. This study explored the role of Ash2l in adipose tissue thermogenesis and obesity-related metabolic dysfunction.MethodsAsh2l was initially identified through transcriptomic analysis, and its expression was further validated in mouse models of high-fat diet (HFD), cold exposure, and CL316,243 stimulation. In vitro gain- and loss-of-function experiments were conducted to assess the role of Ash2l in adipogenesis and thermogenesis. To knockdown Ash2l in vivo, adeno-associated viruses carrying short hairpin RNA targeting Ash2l (AAV-shAsh2l) were injected into either the brown adipose tissue (BAT) or the inguinal white adipose tissue (iWAT). The functional consequences of Ash2l deficiency were evaluated in mice under room temperature, cold exposure, and HFD conditions. Finally, chromatin immunoprecipitation sequencing (ChIP-seq) was employed as an exploratory analysis to identify genomic regions associated with Ash2l during adipocyte development.ResultsOur findings demonstrate that Ash2l modulates the expression of both adipogenic and thermogenic genes in adipocytes. Mice with BAT- or iWAT-knockdown of Ash2l displayed defective cold-induced thermogenesis, aggravated diet-induced obesity, and systemic metabolic dysregulation. Moreover, Ash2l knockdown in BAT under cold exposure or HFD conditions also attenuated thermogenic activity in iWAT, an effect that may be mediated by reduced secretion of FABP4.ConclusionsThese findings establish Ash2l as a critical regulator of adipogenesis and thermogenesis. This study provides important insights into the epigenetic role of Ash2l in maintaining metabolic homeostasis under conditions of nutritional excess.

Ischemic stroke (IS) is a leading cause of mortality and disability worldwide, but effective therapeutic options are limited. In this study, a chemical proteomic strategy is employed using the active compound procyanidin B3 (PB3) as a chemical probe to identify the therapeutic targets for IS. It is discovered that the Ras GTPase-activating protein SH3 domain-binding protein 1 (G3BP1) is a key target of PB3, which exerts a neuroprotective effect by inhibiting the degradation of stress granules and reducing apoptosis. Based on this finding, 14 PB3 derivatives are designed and synthesized, among which compound 6c exhibited potent neuroprotective activity and favorable blood-brain barrier permeability. This study not only establishes G3BP1 as a promising therapeutic target for IS but also highlights the potential of PB3 and its derivatives for the development of IS therapeutic agents.

Hypobaric hypoxia, a defining feature of high-altitude environments, induces significant physiological and pathological changes in the human body. Under the disrupted homeostasis and altered disease microenvironment caused by high-altitude conditions, the immune system exhibits distinct responses compared to those observed in low-altitude settings. Our study investigates the impact of hypobaric hypoxia on autoimmune and autoinflammatory diseases and explores the underlying molecular mechanisms. Using an environmental simulation chamber, we subjected mouse models of experimental autoimmune encephalomyelitis (EAE) and psoriasis to hypobaric hypoxia, simulating conditions at an altitude of 6000 m. Pathological analysis and flow cytometry demonstrated exacerbated disease severity and elevated Th17 cell levels. Given the established role of Th17 cells as key effector cells in autoimmune and autoinflammatory diseases, we further investigated their response through transcriptomic comparisons under normoxic and hypoxic conditions, which identified Activin A as a central regulator of hypoxia-induced Th17 cell differentiation. In mice exposed to hypobaric hypoxia, pharmacological inhibition of Activin A significantly alleviated the severity of psoriasis. Western blotting, flow cytometry, and immunofluorescence analyses confirmed that under normoxic conditions, Activin A stimulation amplified the pathogenic Th17 cell molecular program, whereas blockade of p-PKM2 or ERK signaling suppressed this effect. Collectively, our findings uncover a molecular mechanism whereby hypoxia-triggered Activin A release drives pathogenic Th17 differentiation via the ERK pathway, promoting p-PKM2 nuclear translocation and subsequent transcriptional activation of Th17 master regulators and effector cytokines. This study provides a theoretical foundation for understanding immune dysregulation at high altitudes and offers potential therapeutic strategies for mitigating high-altitude-associated immune disorders.

OBJECTIVE:High-altitude environments are characterized by hypobaric hypoxia and pose a significant physiological challenge. While obesity-related hypoxia is known to impair brown adipose tissue (BAT) function by suppressing lipolysis and thermogenesis, whether hypobaric hypoxia similarly compromises BAT function remains unclear. We hypothesize that, unlike obesity-associated hypoxia, hypobaric hypoxia perturbs BAT thermogenesis via a distinct mechanism involving the upregulation of Plin2. METHODS:Mice were exposed to either normoxia or hypobaric hypoxia (simulating 6000 m, 9.2% O₂) for 30 days. BAT function was assessed by analyzing lipid droplet size, lipid and triglyceride content, and expression of thermogenic protein. Thermogenic capacity was further evaluated in mice subjected to 30 days of normoxia or hypobaric hypoxia, followed by a 2-day cold exposure or 3-day treatment with the β3-adrenergic agonist CL316243. Differential proteomics and in vitro experiments were performed to explore underlying molecular mechanisms. RESULTS:After 30 days of hypobaric hypoxia, mice exhibited reduced body weight (normoxia: 26.64 ± 0.91 vs. hypoxia: 21.94 ± 0.79 g, p = 0.002), yet exhibited enlarged lipid droplets in BAT (142.7 ± 15.52 vs. 387.4 ± 31.91 μm2, p < 0.001) and increased lipid accumulation. Compared with normoxia, hypoxic mice displayed impaired thermogenic responses to both cold exposure and β3-adrenergic stimulation, as indicated by suppressed thermogenic gene expression. Mechanistically, hypobaric hypoxia elevated lactate levels and upregulated PPARγ, which subsequently enhanced Plin2 expression, ultimately leading to defective lipolysis and impaired thermogenesis. CONCLUSIONS:We identify a novel hypoxia-lactate/PPARγ-Plin2 axis that uncouples body weight loss from BAT thermogenesis under hypobaric hypoxia. This pathway represents a previously unrecognized therapeutic target for counteracting metabolic dysfunction induced by high-altitude exposure.

Myocardial hypertrophy is one of the most prominent features of heart failure. SET domain-containing protein 7 (Setd7), a catalytic enzyme responsible for histone H3K4 methylation, has been implicated in various cardiac diseases. In this study we investigated whether Setd7 contributed to the development of cardiac hypertrophy. Male mice were subjected to a hypobaric hypoxic environment for 8 weeks; neonatal rat cardiomyocytes (NRCMs) exposed to hypoxia for 6 h. We showed that hypoxic stimulation significantly upregulated the expression levels of Setd7 along with the expression of hypertrophic markers ANP and BNP in NRCMs. By conducting loss- and gain-of-function assays, we demonstrated that Setd7 modulated the hypertrophic and inflammatory markers in hypoxic cardiomyocytes. We further revealed that Setd7-mediated activation of E2F1 (E2 promoter binding factor 1) triggered the expression of E3 ubiquitin protein ligases WWP2, which catalyzed the ubiquitination and degradation of glutathione peroxidase 4 (GPx4), a critical lipid peroxide-reducing enzyme. This degradation drove extensive lipid peroxidation, thereby exacerbating pathological cardiac hypertrophy. Notably, GPx4 inhibition by ras-selective lethal small molecule 3 (RSL3) abolished the antihypertrophic effects of Setd7 knockdown in cardiomyocytes, underscoring the pivotal role of lipid peroxidation in Setd7-mediated hypertrophic responses. In summary, Setd7 promotes hypoxia-induced cardiac hypertrophy through the Setd7-E2F1-WWP2-GPx4 signaling pathway, suggesting that targeting Setd7 is a promising therapeutic strategy to alleviate hypoxia-induced myocardial hypertrophy.

BACKGROUND:Myocardial infarction (MI)-induced ischemia leads to abnormal ventricular remodeling and cardiac fibrosis, which can ultimately progress to heart failure. Kif23 (kinesin-like protein 23) has been implicated in the progression of various diseases. This study aims to investigate the role of Kif23 in the development of cardiac fibrosis following MI. METHODS:Male C57BL/6J mice received intravenous injections of adeno-associated virus carrying Kif23 shRNA, followed by left anterior descending coronary artery ligation to induce MI. Cardiac function and fibrosis were assessed via echocardiography, histological analysis, and fibrosis marker quantification at 7 and 14 days post-MI. In vitro, adult rat cardiac fibroblasts underwent Kif23 knockdown or overexpression, with subsequent TGF-β1 (Transforming Growth Factor Beta 1) treatment. Proteomic profiling, lipid droplet analysis, and fibrosis/lipid metabolism assessments were performed. RESULTS:Kif23 expression was significantly upregulated in both infarcted myocardium and TGF-β1-stimulated adult rat cardiac fibroblasts. Kif23 knockdown improved cardiac function and attenuated fibrosis post-MI. In vitro, Kif23 silencing suppressed fibroblast proliferation and myofibroblast transdifferentiation, whereas Kif23 overexpression potentiated fibrotic responses. Proteomic profiling identified Ces1d (carboxylesterase 1d) as a key downstream effector upregulated by Kif23 knockdown, concomitant with restored fatty acid β-oxidation. Mechanistically, Kif23 impairs Ces1d-mediated fatty acid β-oxidation via RhoA (Ras Homolog Family Member A)/ROCK1 (Rho-associated protein kinase 1) signaling, driving fibrosis progression. CONCLUSIONS:Our study identifies Kif23 as a novel regulator of post-MI cardiac fibrosis. Mechanistically, Kif23 drives fibroblast proliferation through RhoA activation and exacerbates fibrogenesis by suppressing Ces1d-dependent fatty acid β-oxidation via the RhoA/ROCK1 axis. This metabolic disruption triggers pathological lipid accumulation and myofibroblast transdifferentiation. These findings nominate Kif23 as a promising therapeutic target for antifibrotic interventions.

Introduction Skeletal muscle function is profoundly challenged under high-altitude environments, where hypobaric hypoxia disrupts structural integrity and impairs physiological function. However, few animal studies have examined the impact of hypobaric hypoxia on skeletal muscle and molecular basis. While exercise training holds promise for alleviating hypoxia-induced muscle dysfunction, the understanding of its protective mechanisms remains limited. Objectives We aimed to investigate chronic hypobaric hypoxia-induced myotube atrophy and mitochondrial dysfunction in mouse models and C2C12 cells, and develop a combined exercise strategy (preconditioning and hypoxic training) to mitigate hypoxia-related muscle pathology. Methods A mouse chronic hypobaric hypoxia model (45-day exposure, 6,000 m equivalent) combined with in vitro C2C12 myotube hypoxia simulations was employed. Muscle atrophy, mitochondrial ultrastructure, and molecular pathways were analyzed via histology, proteomics, and functional assays. Exercise interventions included preconditioning (9-week treadmill training) followed by voluntary wheel running under hypobaric hypoxia. Results Chronic hypobaric hypoxia induced pronounced skeletal muscle dysfunction and mitochondrial structural disorganization. However, exercise preconditioning combined with hypoxic training attenuated these hypoxia-induced impairments. Both hypoxic skeletal muscles in vivo and C2C12 cells in vitro exhibited significant Sirt1 downregulation. Notably, overexpression of Sirt1 or treatment with exercise mimetics partially reversed hypoxia-induced myotube atrophy and mitochondrial dysfunction through the PGC-1α/FoxO3a signaling pathway-a mechanism shared with exercise interventions. Conclusion This study uncovers exercise as a potent inducer of hypoxia resilience through Sirt1-dependent mitochondrial repair and multicellular crosstalk (vascular-endothelial-satellite cell axis). Our “train-before-you-climb” approach could transform how we prepare for high-altitude living, offering a drug-free way to keep muscles strong where the air is thin.

Objective: This study aimed to isolate and elucidate the structure of water-soluble constituents from Nelumbo nucifera Gaertn. leaves and evaluate their anti-aging effect. Methods: The natural products were isolated from the 60% aqueous acetone extract of the leaves of N. nucifera by column chromatography on MCI gel CHP 20P, YMC-Gel ODS-AQ-HG, and TSK gel Toyopearl HW-40F. Their structures were elucidated using high-resolution-electrospray ionization-mass spectrometry, one-dimensional and two-dimensional nuclear magnetic resonance spectroscopy, and electronic circular dichroism spectroscopy. The anti-aging activity was evaluated in the Caenorhabditis elegans model. Results/Conclusion: One novel flavonoid nuciferanoid (1), together with eleven known compounds, were isolated from the extract of the leaves of N. nucifera. Among them, compound 1 could be a starting point for further development of anti-aging drugs.

Multi-omics studies of breast ductal carcinoma (BRDC) have advanced the understanding of the disease's biology and accelerated targeted therapies. However, the temporal order of a series of biological events in the progression of BRDC is still poorly understood. A comprehensive proteogenomic analysis of 224 samples from 168 patients with malignant and benign breast diseases is carried out. Proteogenomic analysis reveals the characteristics of linear multi-step progression of BRDC, such as tumor protein P53 (TP53) mutation-associated estrogen receptor 1 (ESR1) overexpression is involved in the transition from ductal hyperplasia (DH) to ductal carcinoma in situ (DCIS). 6q21 amplification-associated nuclear receptor subfamily 3 group C member 1 (NR3C1) overexpression helps DCIS_Pure (pure DCIS, no histologic evidence of invasion) cells avoid immune destruction. The T-cell lymphoma invasion and metastasis 1, androgen receptor, and aldo-keto reductase family 1 member C1 (TIAM1-AR-AKR1C1) axis promotes cell invasion and migration in DCIS_adjIDC (DCIS regions of invasive cancers). In addition, AKR1C1 is identified as a potential therapeutic target and demonstrated the inhibitory effect of aspirin and dydrogesterone as its inhibitors on tumor cells. The integrative multi-omics analysis helps to understand the progression of BRDC and provides an opportunity to treat BRDC in different stages.

TNFα and related inflammatory factor antibody drugs have been orchestrated for the treatment of inflammatory bowel disease (IBD). However, antibody drugs elicited inevitable disadvantages and small molecule drugs are in an urgent need. Herein, we described the discovery, design, synthesis, and SAR studies from furanone glycoside compound Phoenicein (hit) isolated from Chimonanthus salicifolius to D228 (lead). Remarkably, D228 exhibited good inhibitory activity on B and T lymphocyte and excellent anti-IBD efficacy in vivo. Mechanistically, D228 alleviated the inflammation response by downregulating the MyD88/TRAF6/p38 signaling. Importantly, the relationship of D228, Phoenicein, and their aglycone 7a was deduced: D228 could be considered as a prodrug and metabolized to intermediate Phoenicein. In turn, Phoenicein released their shared active aglycone 7a. Additionally, D228 demonstrated good and balanced profiles of safety and efficacy both in vitro and in vivo. These results suggested that D228 could be used as an ideal lead and potentially utilized for IBD chemotherapy.

Endothelial dysfunction is a common complication of diabetes mellitus (DM) and contributes to the high incidence and mortality of cardiovascular and cerebrovascular diseases. Aberrant epigenetic regulation under diabetic conditions, including histone modifications, DNA methylation, and non-coding RNAs (ncRNAs) play key roles in the initiation and progression of diabetic vascular complications. ASH2L, a H3K4me3 regulator, triggers genetic transcription, which is critical for physiological and pathogenic processes. In this study we investigated the role of ASH2L in mediating diabetic endothelial dysfunction. We showed that ASH2L expression was significantly elevated in vascular tissues from diabetic db/db mice and in rat aortic endothelial cells (RAECs) treated with high glucose medium (11 and 22 mM). Knockdown of ASH2L in RAECs markedly inhibited the deteriorating effects of high glucose, characterized by reduced oxidative stress and inflammatory responses. Deletion of endothelial ASH2L in db/db mice by injection of an adeno-associated virus (AAV)-endothelial specific system carrying shRNA against Ash2l (AAV-sh Ash2l ) restored the impaired endothelium-dependent relaxations, and ameliorated DM-induced vascular dysfunction. We revealed that ASH2L expression activated reductase STEAP4 transcription in vitro and in vivo, which consequently elevated Cu(I) transportation into ECs by the copper transporter CTR1. Excess copper produced by STEAP4-mediated copper uptake triggered oxidative stress and inflammatory responses, resulting in endothelial dysfunction. Our results demonstrate that hyperglycemia triggered ASH2L-STEAP4 axis contributes to diabetic endothelial dysfunction by modulating copper uptake into ECs and highlight the therapeutic potential of blocking the endothelial ASH2L in the pathogenesis of diabetic vascular complications.

Oligomeric proanthocyanidins (OPCs) have a variety of biological functions, but the formation of 4,8-interflavan bonds faces scaling-up difficulties due to the challenging control of stereoselectivity and the degree of polymerization. Here we report a process to produce procyanidin B3 (1) by mainly optimizing the condensation reaction and improving benzylation, C4 activation, and one-pot hydrogenolysis reactions. In an optimized seven-step process, the product 1 was achieved by only one-step chromatography in the case of poor crystallinity of polyphenols. This strategy provided effective access to the stereoselective synthesis of the title compound and other C4-C8 connected OPCs.

The aberrant expression of methyltransferase Set7/9 plays a role in various diseases. However, the contribution of Set7/9 in ischemic stroke remains unclear. Here, we show ischemic injury results in a rapid elevation of Set7/9, which is accompanied by the downregulation of Sirt5, a deacetylase reported to protect against injury. Proteomic analysis identifies the decrease of chromobox homolog 1 (Cbx1) in knockdown Set7/9 neurons. Mechanistically, Set7/9 promotes the binding of Cbx1 to H3K9me2/3 and forms a transcription repressor complex at the Sirt5 promoter, ultimately repressing Sirt5 transcription. Thus, the deacetylation of Sirt5 substrate, glutaminase, which catalyzes the hydrolysis of glutamine to glutamate and ammonia, is decreased, promoting glutaminase expression and triggering excitotoxicity. Blocking Set7/9 eliminates H3K9me2/3 from the Sirt5 promoter and normalizes Sirt5 expression and Set7/9 knockout efficiently ameliorates brain ischemic injury by reducing the accumulation of ammonia and glutamate in a Sirt5-dependent manner. Collectively, the Set7/9-Sirt5 axis may be a promising epigenetic therapeutic target.