Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie最新文献

Oxidative stress (OS) is widely recognized as a central promoter to the pathogenesis of neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS) and primary lateral sclerosis (PLS). Cannabis sativa L. synthesizes a complex array of bioactive compounds that extends well beyond the well-known cannabinoids to include a diverse suite of polyphenols, terpenes, fatty acids, tocopherols, and proteins. The non-cannabinoid polyphenolic fraction is composed primarily of flavonoids, stilbenoids, lignans, and lignanamides, which contribute substantially to the plant's antioxidant, anti-inflammatory, and neuroprotective properties. This study investigates the redox-modulating and cytoprotective properties of a polyphenolic fraction derived from Cannabis sativa L. in SH-SY5Y neuroblastoma cells. Neurons were treated with various concentrations of the aqueous polyphenolic cannabis extract and exposed to oxidative stress using hydrogen peroxide (100 µM). Protein and gene expression related to redox signalling were analyzed via Western blot and qPCR, and molecular docking studies were performed in silico. Furthermore, antioxidant enzymes activity was measured by spectrophotometry. Results revealed that the phenolic fraction significantly activated the Keap1/Nrf2 pathway, increased expression of PRDX1 and PRDX3, and enhanced endogenous antioxidant defences. Simultaneously, it reduced endoplasmic reticulum stress-induced apoptosis (via Bax/Bcl-2 modulation) and attenuated inflammatory markers, including NO, NF-κB2, IL-6, and IL-8. In silico docking studies identified Leu583 as a key residue in Nrf2-ligand interactions. These findings suggest that Cannabis sativa L. polyphenols are key bioactive compounds modulating redox homeostasis and inflammation, and offering neuroprotective benefits with potential relevance in diseases involving mitochondrial dysfunction and oxidative damage.

5-Fluorouracil (5-FU) is a first-line chemotherapeutic for colorectal cancer (CRC), yet its clinical application is limited by low bioavailability, rapid systemic clearance, and off-target toxicity. We developed tunable PEGylated bovine serum albumin (BSA) nanoparticles as a nanocarrier platform for 5-FU delivery, aiming to improve drug encapsulation, enhance cytotoxic selectivity toward CRC cells, and modulate oxidative and immune-related responses. Using PEG of varying molecular weights (2, 4, and 10 kDa), monodisperse nanoparticles (71-111 nm) were synthesized via a desolvation method and characterized by multi-angle dynamic light scattering, transmission electron microscopy, microscale thermophoresis and HPLC. PEGylation improved colloidal stability and increased 5-FU loading efficiency, particularly at higher drug input concentrations. Binding studies revealed molecular weight- and concentration-dependent interactions of PEG and 5-FU with albumin, influencing nanoparticle architecture and physicochemical properties. Biological evaluation was performed across CRC cell lines (HT-29, Caco-2), non-cancerous fibroblasts (L929), and THP-1-derived macrophages. PEGylated BSA-FU nanoparticles exhibited selective, long-term cytotoxicity toward colorectal cancer cells. In macrophages, PEGylated formulations modulated phagocytic activity and oxidative stress responses in a formulation-dependent manner, indicating that PEGylation alters-rather than uniformly suppresses-immune cell interactions. These findings highlight the importance of rational PEG architecture optimization and provide a strong foundation for future in vivo studies addressing repeated dosing, immune interactions, and translational potential in colorectal cancer therapy.

Osteoporosis is a chronic disorder marked by bone wasting and increased bone fragility. Targeted inhibition of osteoclastogensis is currently the core therapeutic strategy. Hexahydrocurcumin (HHC), derived from Zingiberis Rhizoma, has been shown to exhibit anti-inflammatory and antioxidant properties; however, its effects on osteoclasts regulation and osteoporosis pathogenesis remain unexplored. We conducted this study to observe the influence of HHC on RANKL-mediated osteoclast precursor differentiation and OVX-dependent osteoporotic mice. In this study, we revealed that HHC significantly attenuated the generation and bone resorptive function of osteoclasts induced by RANKL in vitro, which was achieved by targeting and inhibiting the phosphorylation of c-Src, a critical molecule in osteoclast differentiation. Next, HHC inhibited the subsequent Ca²⁺ influx and NFATc1 nuclear translocation, thereby suppressing the expression of osteoclastogenic regulators such as Acp5 and Mmp9. Furthermore, we validated that HHC inhibits osteoclastogenesis by targeting c-Src through siRNA-mediated silencing of c-Src. In the in vivo study, HHC notably alleviated bone loss in OVX-dependent osteoporotic mice. These findings suggest that HHC alleviates osteoporosis by inhibiting osteoclastogenesis via targeting c-Src, which provide preliminary evidence for the potential of HHC for the treatment of osteoporosis.

Sexually transmitted infections (STIs) remain a major global health concern, contributing significantly to morbidity and facilitating the co-transmission of other pathogens. Recent outbreaks of Monkeypox virus (MPXV) have further underscored the urgent need for broad-spectrum antiviral agents effective against emerging and re-emerging sexually transmissible viruses. We report here the design and synthesis of a series of 3-cyanoquinoline-based Src inhibitors to evaluate their antimicrobial efficacy against sexually transmitted pathogens. Among them, compound 7d demonstrated potent inhibitory activity against MPXV, Herpes simplex virus types 1 and 2, Hepatitis C virus, Human immunodeficiency virus, and Chlamydia trachomatis at non-toxic concentrations. Owing to its broad-spectrum profile and favorable cytotoxicity profile, compound 7d represents a promising candidate for development as a topical microbicide for the prevention and treatment of STIs. Interestingly, the screening also identified compound 7g, which, despite lacking Src inhibitory activity, exhibited selective antiviral activity against members of the Poxviridae family, suggesting the involvement of alternative host-dependent mechanisms that can be further exploited. Both compounds were non-toxic in relevant epithelial and mucosal tissue models. Collectively, these findings highlight the therapeutic potential of 3-cyanoquinoline derivatives as scaffolds for the development of novel broad-spectrum microbicides targeting a range of sexually transmitted pathogens.

Radioligand therapy targeting prostate-specific membrane antigen (PSMA) has demonstrated promising clinical outcomes for patients with metastatic castration-resistant prostate cancer. However, accumulation in non-target organs can lead to significant radiotoxicity, affecting patient well-being and potentially requiring treatment discontinuation. Ultrasound combined with microbubbles (USMB) has been shown to transiently permeabilize biological barriers, enabling efficient and safe drug delivery to tumor tissues while minimizing dose-limiting toxicities. This study explores the impact of different conditions of USMB on the distribution of the diagnostic radiopharmaceutical [¹⁸F]F-PSMA-1007 as a preliminary step before applying therapeutic radiopharmaceuticals (RPs) in a preclinical subcutaneous model. Immunodeficient mice bearing human LNCaP tumors were treated with different ultrasound parameters (i.e., pulse length and pressure). Each mouse received an intravenous injection of [¹⁸F]F-PSMA-1007 (5.1 ± 1.7 MBq) and was imaged by PET/CT 2 h post-injection (p.i.). Additionally, an intravenous injection of TRITC Dextran (100 µL, 70 kDa, 5 mg/mL) was administered to quantify its extravasation into the tumor, correlated with PSMA and CD31 expression via immunofluorescence. Contrast-enhanced ultrasound imaging was also performed to assess tumor perfusion. Results showed a mild, though non-significant, trend toward increased [18 F]F-PSMA-1007 in most groups compared to the control, except for those exposed to short pulses associated with high-pressure. These findings highlight the potential of USMB to enhance drug delivery for PSMA uptake but also underscore the necessity for careful consideration of ultrasound parameters to prevent tissue damage.

Diabetes is a major risk factor for diabetic encephalopathy (DE), which is closely associated with sporadic Alzheimer's disease. Folic acid (FA) receptor signaling can suppress generation of neuropathogenic amyloid-beta (Aβ) induced by high extracellular glucose, suggesting that enhanced activation of this pathway could be a therapeutic strategy against DE-associated dementia, but the precise molecular signaling mechanisms are unclear. We report that high glucose levels increased the expression of amyloid precursor protein (APP) and β-secretase (BACE1) in cultured neurons and concomitantly induced amyloidogenesis, while FA treatment suppressed high glucose-stimulated expression of APP and BACE1, Aβ release, and accumulation of mitochondrial reactive oxygen species. Expression of nuclear factor erythroid 2-related factor 2 (Nrf2) was minimal under high glucose conditions, but was significantly upregulated together with downstream antioxidant enzymes following FA co-treatment. High glucose stimulation also increased folate receptor 1 (FOLR1) mRNA expression, suggesting a compensatory protective response. While treatment with 5-methyltetrahydrofolate (5-MTHF), the activated form of folate, did not significantly alter high glucose-induced upregulation of APP and BACE1, knockdown of FOLR1 mRNA reduced high glucose-stimulated Nrf2 expression and further augmented APP and BACE1 expression under high glucose conditions. Treatment with the STAT3 inhibitor 5'15-DPP also abolished high glucose-stimulated Nrf2 expression and increased APP and BACE1 expression levels. These findings indicate that FA/FOLR1 activation suppresses high glucose-induced amyloidogenesis by mitigating mitochondrial oxidative stress via STAT3/Nrf2 pathway signaling. In conclusion, present study suggests that the FA/FOLR1/STAT3/Nrf2 pathway is an effective therapeutic target for DE.

Platelets, which mediate hemostasis, have been implicated in cardiovascular diseases such as myocardial infarction and stroke. Calycosin, a flavonoid extracted from the root of Astragalus membranaceus, has diverse biological effects, including anticancer, anti-inflammatory, and antidiabetic effects. Whether calycosin inhibits platelet activation and thrombus formation is unclear. The present study explored the mechanisms underlying the potential antiplatelet and antithrombotic effects of calycosin. Platelet aggregation assays, flow cytometry, and Western blotting were performed to analyze the antiplatelet effects of calycosin. Thrombus formation in mouse mesenteric vessels was investigated to analyze the antithrombotic effects of calycosin. Calycosin selectively inhibited collagen-induced platelet aggregation and glycoprotein VI-mediated downstream signaling, including pathways involving phospholipase Cγ2 and protein kinase C. Additionally, calycosin attenuated the activation of protein kinase B and mitogen-activated protein kinase and further suppressed collagen-induced granule release, calcium mobilization, and glycoprotein IIb/IIIa activation. In vivo experiments revealed that calycosin prevented pulmonary thromboembolism and delayed thrombus formation in mouse mesenteric vessels, without affecting hemostasis. This study is the first to demonstrate that calycosin effectively prevents platelet activation and thrombus formation, partly by targeting glycoprotein VI-mediated signaling, without affecting hemostasis. These findings highlight the therapeutic potential of calycosin for cardiovascular diseases.

The COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has underscored the need for broad and potent antiviral agents. Although overall disease severity has diminished, the persistent risk of reinfection highlights the continued demand for novel therapeutic options. In this study, we performed an image-based high-throughput screening campaign of 11,030 small molecules-including nucleoside analogs, known antivirals, and diverse bioactives-to identify inhibitors of SARS-CoV-2 infection. Using an immunofluorescence assay that quantified viral proteins and assessed cell viability by Hoechst nuclear staining, we identified 97 primary hits in infected Vero cells. Dose-response evaluation confirmed 18 compounds active against both ancestral and Omicron variants, and subsequent validation in human lung cell lines (A549-hACE2-TMPRSS2 and Calu-3) highlighted multiple cysteine protease inhibitors as strong antiviral candidates. Among these, MG-101 emerged as a potent cysteine protease inhibitor with favorable pharmacokinetics, metabolic stability, and robust in vivo antiviral efficacy. Docking analysis and enzymatic assays demonstrated that MG-101 inhibits the SARS-CoV-2 3CL protease, and combination studies revealed in vitro synergistic antiviral activity with remdesivir. Together, these findings establish MG-101 as a potential therapeutic lead for COVID-19 and illustrate the value of image-based high-throughput screening for accelerating antiviral drug discovery.

Breast cancers are characterized by complex energy metabolisms involving the Warburg effect but also mitochondria, although this area is not yet well understood. Tumor cells are particularly flexible by choosing oxidative phosphorylation (OXPHOS) or glycolysis depending on the needs and aggressiveness. Within the mitochondria, a HSP90-chaperone protein, TRAP1, exerts regulatory effects on several vital functions such as OXPHOS, production of reactive oxygen species and apoptosis by interacting with members of the respiratory chain or the mPTP. However, not all of its roles have yet been elucidated. Here, we propose to modulate TRAP1 functions using a mitochondriotropic molecule (containing triphenylphosphonium) targeting its C-terminal domain, 6BrCaQ-C₁₀-TPP, in breast tumor cells. Its blocks proliferation with no massive apoptosis, after 24 h of treatment, and induces dissipation of the mitochondrial membrane potential. 6BrCaQ-C₁₀-TPP also appears to modulate regulators of epithelial-mesenchymal transition (Snail and ZEB1) without a common response in all cell lines. Furthermore, the chaperone machinery is affected with a decrease of HSF1 and HSP70, but without degradation of HSP90 or TRAP1, while decreasing the levels of SDH-A and/or SDH-B, partner of TRAP1. Finally, short-term treatments (1 and 3 h) with 6BrCaQ-C₁₀-TPP modify energy metabolism by promoting glycolysis. In conclusion, modulation of TRAP1 on the C-terminal domain by 6BrCaQ-C₁₀-TPP exerts a cell-line dependent anti-tumor effect by modulating major mitochondrial functions in vitro. The differences between cell types need to be clarified. This study confirms that TRAP1 is a target of interest in breast cancer cells, but some of its functions still need to be elucidated.

Endothelial dysfunction is a hallmark of type 2 diabetes mellitus (T2DM) and a major contributor to cardiovascular complications. Although glucagon-like peptide-1 receptor agonists (GLP-1RAs) improve glycemic control and cardiovascular outcomes, the mechanisms linking GLP-1RA therapy, gut microbiome modulation, and endothelial function remain incompletely understood. In this study, we investigated whether the GLP-1RA liraglutide improves endothelial dysfunction in T2DM through microbiome-associated mechanisms that support vascular homeostasis. Male db/db mice and non-diabetic controls were treated with liraglutide (300 μg/kg/day, intraperitoneally) or saline for two weeks. Vascular function was assessed in mesenteric resistance arteries using wire myography. Human umbilical vein endothelial cells (HUVECs) were exposed to high glucose with or without liraglutide or the short chain fatty acid (SCFA), butyrate. Endothelial nitric oxide (NO) signaling was evaluated by eNOS (at Ser1177) phosphorylation and nitrite production. Gut microbiota composition was analyzed by 16S rRNA gene sequencing. Liraglutide significantly improved endothelium-dependent relaxation in db/db mice and restored high glucose-induced impairment of eNOS phosphorylation and NO production in HUVECs. In vivo, diabetes was associated with marked gut dysbiosis characterized by reduced alpha diversity and depletion of SCFA-producing taxa. Liraglutide treatment substantially restored microbial diversity and enriched beneficial genera, including Lachnospiraceae and Lactobacillus. Consistently, low-dose butyrate modestly enhanced NO production in endothelial cells. These findings support the concept of a GLP-1RA-microbiome-vascular axis, in which liraglutide-associated remodeling of the gut microbiota may contribute to improved endothelial NO signaling and vascular function in diabetes.