Elevated ROS Research Articles

AZD6738 Attenuates LPS-Induced Corneal Inflammation and Fibrosis by Modulating Macrophage Function and Polarization.

This study aimed to evaluate the therapeutic potential of AZD6738, an ATR inhibitor, in LPS-induced bacterial keratitis (BK) by targeting macrophage function and polarization. A murine model of LPS-induced BK was established, with AZD6738 (100µM) administered subconjunctivally and topically. Corneal opacity, edema, and inflammation were assessed using slit-lamp microscopy and histological analysis. Macrophage infiltration and fibrosis were evaluated via immunofluorescence, qPCR, and Western blotting. In vitro, RAW264.7 cells were treated with 2.5µM AZD6738 to examine its effects on cell viability, oxidative stress, and inflammation-related gene expression. AZD6738 significantly reduced corneal opacity, thickness, and neovascularization in LPS-treated mice. It suppressed macrophage infiltration, collagen deposition, and pro-inflammatory cytokine expression. In RAW264.7 cells, AZD6738 induced cell death, elevated ROS production, and downregulated inflammatory markers. ATR inhibition mitigated NF-κB activation and modulated macrophage polarization, attenuating M1 pro-inflammatory responses. AZD6738 effectively alleviates LPS-induced corneal inflammation and fibrosis by regulating macrophage function and polarization via the NF-κB signaling pathway. ATR inhibition represents a promising therapeutic strategy for the treatment of corneal inflammation.

Lithium enhanced plasmid-mediated conjugative transfer of antimicrobial resistance genes in Escherichia coli: Different concentrations and mechanisms.

Conjugative transfer, a pivotal mechanism in the transmission of antimicrobial resistance genes, is susceptible to various environmental pollutants. As an emerging contaminant, lithium (Li) has garnered much attention due to its extensive applications. This research investigated the effects of Li on conjugative transfer process, examining biochemical and omics perspectives. Results revealed that Li could increase the conjugative transfer frequency of both donor and recipient via different mechanisms at varying concentrations. At 0.1 mg/L LiCl, a notable increase in conjugative transfer frequency occurred without ROS elevation. However, the surge of ROS was identified as a crucial regulator at 100 mg/L LiCl, as eliminating ROS would significantly decrease the conjugative transfer frequency. Besides, comparative transcriptome analysis revealed consistent variations in "SOS response", "quorum sensing" and "oxidative phosphorylation" pathways at both 0.1 mg/L and 100 mg/L LiCl concentrations, suggesting their pivotal roles as targets for Li regulation and is independent of Li concentration. While genes related to "conjugative transfer", "pili", "outer membrane protein" and "antioxidant enzyme" were only significantly regulated by 100 mg/L LiCl, possible to be the specific reasons for High (100 mg/L) LiCl increased conjugative transfer frequency. This study reveals the distinct effects and mechanisms of different concentration of Li on conjugative transfer in E. coli, providing a theoretical basis for the understanding of the environmental effects of Li.

Curcumin chemo-sensitizes intrinsic apoptosis through ROS-mediated mitochondrial hyperpolarization and DNA damage in breast cancer cells.

Curcumin (CUR), a polyphenol phyto-compound extracted from turmeric rhizome (Curcuma longa), suppresses cancer by inducing apoptosis while also limiting cell survival and proliferation. This in vitro and in silico research work focuses on the synergistic sensitization of Doxorubicin (DOXO) on regulating ROS-mediated apoptosis of the breast cancer cells (MDA-MB-231 and MCF-7) in DOXO-CUR co-treatment. We observed dose-dependent cytotoxicity, increased ROS production, and mtDNA fragmentation by reduced membrane potential. The combined molecular docking of Bcl2, Bax, and Caspase3 proteins with DOXO and CUR with lower binding energies proves the stable interactions of protein-ligand complexes in the combination doses. Cell survival was measured by MTT and flow cytometry assays. Mitochondrial ROS production, mtDNA condensation, and MMP depletion were documented using fluorescence micrographs. The enrichment analysis of the ROS pathway genes by RT-qPCR (relative fold change) indicates the activation of Caspase3-mediated intrinsic apoptosis. Autodock 4.2 and Gromac 2022.4 were performed for in silico binding interaction and stability analysis. Our study calculates the DOXO and CUR combination (0.33 + 33 μM in MDA-MB-231 and 0.14 + 14 μM in MCF-7) shows maximum growth inhibition (70-75 %) by elevated oxidative stress and reduced membrane potential, which suggests that CUR could be a potential therapeutic agent for treating breast cancers in near future. The method of apoptosis was further analyzed, where elevated cellular ROS level by CUR + DOXO combination therapy depleted mitochondrial membrane potential and enhanced the DNA condensation. The mitochondrial pro-apoptotic genes BAX, BAK, BIM, CASPASE9, and CASPASE3 and anti-apoptotic BCL2 gene expressions depicted triggering intrinsic apoptosis pathway, co-relating with the in silico molecular docking, simulation, and MM-PBSA energy calculations. The synergism between CUR and DOXO was also validated by increased binding affinity and reduced inhibitory constant against key proteins Bcl2, Bax, and Caspase3. Bcl2-DOXO showed BE: -5.03 and Bcl2-DOXO-CUR showed BE: -4.7. Whereas, Bax-DOXO binding energy was -5.49 and Bax-DOXO-CUR binding was -3.83. The most preferable synergistic binding was found with Caspase3 protein, where Caspase3-DOXO docking energy was -1.63 but Caspase3-DOXO-CUR combined docking energy was -3.51. The stability of protein-ligand complexes was accessed with MD simulations and binding free energy calculations, Bcl2-CUR-DOXO combination complex showed ∆G: -25.62, Bax-DOXO-CUR complex showed the maximum ∆G: -34.18, with Caspase-CUR-DOXO complex (∆G: -15.18), indicating the proteins most stable conformation while interacting with CUR + DOXO combination. The correlation between the in vitro and in silico analysis of apoptosis pathway components widens the further research scope for proteomics and organoid studies, which might be worthy for successful clinical trials.

Targeting p62 by sulforaphane promotes autolysosomal degradation of SLC7A11, inducing ferroptosis for osteosarcoma treatment.

Osteosarcoma (OS) is the most prevalent malignant bone tumor in children and adolescents worldwide. Identification of novel therapeutic targets and development of targeted drugs are one of the most feasible strategies for OS treatment. Ferroptosis, a recently discovered mode of programmed cell death, has been implicated as a potential strategy for cancer therapy. Sulforaphane (SFN), the main bioactive compound derived from cruciferous vegetables, has shown potential anti-cancer effects with negligible toxicity. However, the role of ferroptosis in the effect of SFN on OS remains unknown. In the present study, we found that SFN acted as a potent ferroptosis inducer in OS, which was demonstrated by various inhibitors of cell death. The SFN-induced ferroptotic cell death was characterized by elevated ROS levels, lipid peroxidation, and GSH depletion, which was dependent on decreased levels of SLC7A11. Mechanically, SFN directly targeted p62 protein and enhanced p62/SLC7A11 protein-protein interaction, thereby promoting the lysosomal degradation of SLC7A11 and triggering ferroptosis. Notably, both subcutaneous and intratibial OS models in nude mice confirmed the ferroptosis associated anti-cancer efficacy of SFN in vivo. Hence, our findings demonstrate that SFN exerts its anti-cancer effects through inducing SLC7A11-dependent ferroptosis in OS, providing compelling evidence for the application of SFN in OS treatment.

Ovarian premature aging: VIP as key regulator of fibro-inflammation and foamy macrophages generation.

Ovarian aging is associated with fibro-inflammation, contributing to the decline in oocyte count and quality. Given the immunomodulatory properties of the vasoactive intestinal peptide (VIP) in the reproductive tract, we investigated its role in maintaining ovarian immune homeostasis and preventing premature aging. We evaluated young VIP knockout (KO) mice, comparing them to young wild type (WT) females, for signs of premature aging. Histological staining revealed aberrant ovarian morphology in VIP KO mice, characterized by increased atretic follicles and decreased ovarian reserve compared to WT controls. Moreover, VIP KO ovaries showed reduced vascularization, increased collagen deposition and elevated ROS and IL-1β levels. Foamy macrophages were significantly predominant, indicating premature aging in young VIP KO ovaries. To determine potential mechanisms behind these pathogenic changes, we conditioned peritoneal macrophages from young WT or VIP KO mice in vitro with ovarian-conditioned media from young WT or VIP KO mice to mimic the respective ovarian microenvironment. When WT or VIP KO peritoneal macrophages were conditioned with ovarian media from their respective genotypes, lipid droplet accumulation increased compared to control medium. In cross-genotype experiments, WT macrophages conditioned with media from VIP KO ovaries selectively accumulated higher levels of lipid droplets, whereas no differences were observed in VIP KO macrophages conditioned with WT ovarian media. This suggests that VIP KO macrophages are uniquely sensitized to the inflammatory environment of VIP KO ovaries, implicating both ovarian factors and macrophage status. These findings highlight the role of VIP in preventing fibro-inflammation, thereby preserving ovarian health and preventing premature aging.

Lysosomes finely control macrophage inflammatory function via regulating the release of lysosomal Fe2+ through TRPML1 channel

Lysosomes are best known for their roles in inflammatory responses by engaging in autophagy to remove inflammasomes. Here, we describe an unrecognized role for the lysosome, showing that it finely controls macrophage inflammatory function by manipulating the lysosomal Fe2+—prolyl hydroxylase domain enzymes (PHDs)—NF-κB—interleukin 1 beta (IL1B) transcription pathway that directly links lysosomes with inflammatory responses. TRPML1, a lysosomal cationic channel, is activated secondarily to ROS elevation upon inflammatory stimuli, which in turn suppresses IL1B transcription, thus limiting the excessive production of IL-1β in macrophages. Mechanistically, the suppression of IL1B transcription caused by TRPML1 activation results from its modulation on the release of lysosomal Fe2+, which subsequently activates PHDs. The activated PHDs then represses transcriptional activity of NF-κB, ultimately resulting in suppressed IL1B transcription. More importantly, in vivo stimulation of TRPML1 ameliorates multiple clinical signs of Dextran sulfate sodium-induced colitis in mice, suggesting TRPML1 has potential in treating inflammatory bowel disease.

Methyl 3-Bromo-4,5-dihydroxybenzoate Attenuates Inflammatory Bowel Disease by Regulating TLR/NF-κB Pathways.

Inflammatory bowel disease (IBD) is characterized by uncontrolled, chronic relapsing inflammation in the gastrointestinal tract and has become a global healthcare problem. Here, we aimed to illustrate the anti-inflammatory activity and the underlying mechanism of methyl 3-bromo-4,5-dihydroxybenzoate (MBD), a compound derived from marine organisms, especially in IBD, using a zebrafish model. The results indicated that MBD could inhibit the inflammatory responses induced by CuSO4, tail amputation and LPS in zebrafish. Furthermore, MBD notably inhibited the intestinal migration of immune cells, enhanced the integrity of the gut mucosal barrier and improved intestinal peristalsis function in a zebrafish IBD model induced by trinitro-benzene-sulfonic acid (TNBS). In addition, MBD could inhibit ROS elevation induced by TNBS. Network pharmacology analysis, molecular docking, transcriptomics sequencing and RT-PCR were conducted to investigate the potential mechanism. The results showed that MBD could regulate the TLR/NF-κB pathways by inhibiting the mRNA expression of TNF-α, NF-κB, IL-1, IL-1β, IL6, AP1, IFNγ, IKKβ, MyD88, STAT3, TRAF1, TRAF6, NLRP3, NOD2, TLR3 and TLR4, and promoting the mRNA expression of IL4, IκBα and Bcl-2. In conclusion, these findings indicate that MBD could be a potential candidate for the treatment of IBD.

Lantana camara L. induces a multi-targeted cell death process in Leishmania amazonensis. R1

Lantana camara L. is a species known for its broad spectrum of bioactivities and is commonly used in folk therapy to address inflammatory, dermatological, gastrointestinal, intestinal worms and protozoan diseases. It boasts a diverse array of secondary metabolites such as terpenes, flavonoids, and saponins. However, despite its rich chemical profile, there remains a scarcity of studies investigating its antileishmanial properties. This research aims to explore the antileishmanial potential of L. camara, focusing also on its mechanism of action against Leishmania amazonensis. The ethanolic extract of L. camara leaves (LCE) was obtained through static maceration, and its phytoconstituents were identified using UFLC-QTOF-MS. The colorimetric MTT method was conducted to determine the effect of LCE on promastigotes of L. amazonensis and murine macrophages. The anti-amastigote activity was evaluated by counting intracellular parasites in macrophages after Giemsa staining. Additionally, investigations into the mechanisms underlying its action were conducted using cellular and biochemical approaches. LCE exhibited significant activity against both promastigotes and intracellular amastigotes of L. amazonensis, with IC50 values of 12.20μg/mL ± 0.12 and 7.09μg/mL ± 1.24, respectively. These IC50 values indicate very promising antileishmanial activity, comparable to those found for the positive control miltefosine (5.10μg/mL±1.79 and 8.96μg/mL±0.50, respectively). Notably, LCE exhibited negligible cytotoxicity on macrophages (IC50=223.40μg/mL±47.02), demonstrating selectivity towards host cells (SI=31.50). The antileishmanial activity of LCE involved a multi-targeted cell death process, characterized by morphological and ultrastructural alterations observed through SEM and TEM analyses, as well as oxidative effects evidenced by the inhibition of trypanothione reductase, elevation of ROS and lipid levels, and mitochondrial dysfunction evaluated using DTNB, H2DCFDA, Nile red, and JC-1 assays. Additionally, extraction of ergosterol and double labeling with annexin V and PI revealed modifications to the organization and permeability of the treated parasite's plasma membrane. LCE was found to consist predominantly of terpenes, with lantadenes A, B, and C being among the eleven compounds identified through UFLC-QTOF-MS analysis. The extract of L. camara presents a diverse array of chemical constituents, prominently featuring high terpene content, which may underlie its antileishmanial properties through a combination of apoptotic and non-apoptotic mechanisms of cell death induced by LCE. This study underscores the therapeutic potential of L. camara as a candidate for antileishmanial treatment, pending further validation.

T-2 toxin triggers immunotoxic effects in goats by inducing ferroptosis and neutrophil extracellular traps.

T-2 toxin, a prevalent mycotoxin, represents a notable global public health risk. Neutrophil extracellular traps (NETs) and ferroptosis are involved in a variety of pathophysiological processes and are implicated in goat immunity. However, the impact of T-2 toxin on NETs release, ferroptosis, and their interplay have not been previously documented. In this study, neutrophils were stimulated with T-2 toxin for 4h. The structure and mechanism of NETs were analyzed using immunofluorescence and Pico Green staining. The expressions of glutathione peroxidase 4 (GPX4) and ferritin (FT) was quantified by qRT-PCR and western blotting. The levels of ROS and lipid ROS were assessed using DCFH-DA and C11 BODIPY 581/591 probes, and cellular mitochondria Fe2+ were detected by using Mito-FerroGreen probe. Inhibitors were utilized to explore the interaction between these two processes. The results confirmed that the T-2 toxin stimulated the NETs production, characterized by a structure co-modified by citrullinated histones (citH3), neutrophil elastase (NE) and DNA. Notably, significant inhibition of NETs production by T-2 toxin was observed with the NOX inhibitor DPI (P<0.001), the ERK inhibitor U0126 (P<0.001), the TLR2 inhibitor C29 (P<0.001), and the TLR4 inhibitor TLR4-IN-C34 (P<0.001). T-2 toxin triggered ferroptosis in neutrophils by suppressing GPX4 and FT expression, elevating ROS and lipid ROS, and augmenting the concentration of mitochondrial Fe2+. The ferroptosis inhibitor Fer-1 could rescue this induction; however, Fer-1 was unable to inhibit NETs which is induced by T-2 toxin. Conversely, T-2 toxin effectively triggered the downregulation of GPX4, which was counteracted by DPI, U0126, C29, and C34. This research elucidates the immunotoxic mechanisms of T-2 toxin in goat neutrophils and offers a novel perspective on preventing and treating T-2 toxin.

Complex Immunotoxic Effects of T-2 Toxin on the Murine Spleen and Thymus: Oxidative Damage, Inflammasomes, Apoptosis, and Immunosuppression

T-2 toxin (T-2), a highly stable and toxic mycotoxin, poses a significant public health risk as an inevitable environmental pollutant. However, the mechanisms behind its immunotoxic and immunosuppressive effects are not fully understood. For this study, sixty healthy 4-week-old male C57BL/6N mice were divided randomly into four groups and treated for 28 days with T-2 concentrations of 0, 0.5, 1.0, and 2.0mg/kg. Our findings revealed significant damage to the thymus and spleen that was proportional to the dose administered, as evidenced by changes in organ indices and histopathological abnormalities. We observed mitochondrial swelling, chromatin condensation, and nuclear structure disruptions in these organs. Even at low doses (0.5mg/kg), T-2 administration resulted in significant immunosuppression, as evidenced by disturbed blood parameters and altered CD4+/CD8+ ratios. Elevated ROS and MDA levels indicate oxidative damage, whereas SOD, T-AOC, CAT, and GSH levels are reduced in both the thymus and spleen. Furthermore, the levels of NLRP3, ASC, caspase-1, and IL-1β proteins were significantly elevated, indicating the activation of the NLRP3 inflammasome pathway. Additionally, activation of the apoptosis pathway was demonstrated by an increased Bax/Bcl-2 ratio and heightened activation of caspase-3 and -9. Transcriptomic analysis elucidated the pivotal role of mitochondrial pathways in T-2-induced immunotoxicity. This study elucidates the significant immunotoxic effects of T-2 on the murine spleen and thymus, detailing the underlying mechanisms of T-2-induced immunosuppression. The key mechanisms identified include oxidative stress, activation of the NLRP3 inflammasome, apoptosis, and mitochondrial dysfunction. These findings reveal critical pathways through which T-2 impairs immune system functionality and provide a basis for developing targeted therapeutic strategies to mitigate its immunotoxic effects.

Docosahexaenoic Acid Inhibits p62‐Dependent Autophagy by Targeting HSP70A1A/TGM‐2 Axis to Alleviate Arecoline‐Induced Oral Submucosal Fibrosis

Background: The role of docosahexaenoic acid (DHA) in fibrosis of other organs has been studied, but its function in oral submucous fibrosis (OSF) has not been reported. This study aimed to investigate the role and mechanism of DHA in OSF.Methods: OSF rat and cell models were established induced by arecoline. Through a series of in vivo and in vitro experiments, the function of DHA in OSF was investigated. Mechanistically, the interaction of TGM‐2 with HSP70A1A and p62 proteins was validated using co‐immunoprecipitation. Additionally, in cells transfected with overexpression vectors of HSP70A1A or TGM‐2 and treated with DHA and arecoline or co‐treated with a p62 inhibitor XRK3F2 along with DHA and arecoline, the function of the DHA/HSP70A1A/TGM‐2/p62 axis in OSF was explored.Results: In vivo, arecoline caused severe pathological damage and fibrosis in rat oral mucosal tissues and induced overexpression of HSP70A1A. Arecoline treatment also elevated tissue ROS levels and the expression of α‐SMA, Collagen I, TGM‐2, and LC3 II/I, while decreasing tissue p62 protein expression and serum GSH levels. Treatment with DHA reversed these changes and improved the pathological damage and fibrosis in OSF rats. In vitro, arecoline induced the expression of HSP70A1A in a concentration‐dependent manner, and DHA inhibited its expression by directly binding to HSP70A1A and reducing the expression of α‐SMA, Collagen I, TGM‐2, LC3 II/I, and ROS levels induced by arecoline in cells, while increasing p62 protein expression and GSH levels in cell supernatants. Furthermore, arecoline induced TGM‐2 expression, and overexpression of HSP70A1A counteracted the protective effect of DHA on cells and the suppression of TGM‐2 expression. TGM‐2 interacted with HSP70A1A and p62 proteins. Overexpression of TGM‐2 or treatment with XRK3F2 activated autophagy and abolished the protective effect of DHA on cells.Conclusion: DHA inhibits p62‐dependent autophagy through targeting the HSP70A1A/TGM‐2 axis, thereby alleviating arecoline‐induced OSF. These results suggest that DHA and its mediated autophagy regulation mechanism can be a therapeutic target for OSF.

β-pinene ameliorates ICV-STZ induced Alzheimer's pathology via antioxidant, anticholinesterase, and mitochondrial protective effects: In-silico and in-vivo studies.

Alzheimer's disease (AD) is a leading cause of dementia, characterized by progressive neurodegeneration and cognitive dysfunction. The disease aetiology is closely associated with proteinopathies, mitochondrial abnormalities, and elevated ROS generation, which are some of the primary markers for AD brains. The current research was intended to elucidate the chemical interaction of β-pinene against potential targets and evaluate its neuroprotective potential in ICV-STZ-induced sAD. ology: The potential binding interactions of β-pinene and galantamine were evaluated against the active sites of PP2A, SOD1, catalase-3, and AChE using Autodock vina. Additionally, the β-pinene and galantamine were subjected to tests of their ADMET by employing the Swiss ADME and Protox-II web servers. To assess the neuroprotective potential, β-pinene (50, 100, and 200 mg/kg) and galantamine (2 mg/kg) was administered p.o in ICV-STZ-treated wistar rats for 21 days. Moreover, behavioral parameters (NOR & MWM), biochemical, AChE activities, and mitochondrial complexes were performed. A molecular docking study showed that β-pinene can interact with human PP2A, SOD1, Catalase-3, and AChE with better ligand efficiency as compared to galantamine. In-vivo data showed that β-pinene treatment (100, and 200 mg/kg) for 21 days exhibited significantly enhanced cognitive performance, as shown in behavioral studies. Additionally, β-pinene treatment significantly re-established antioxidant levels and mitochondrial capacities and attenuated altered AChE activity as compared to ICV-STZ-induced groups. In-silico studies revealed that β-pinene shared the same binding pocket as galantamine, supporting its neuroprotective effects in the ICV-STZ-induced animal model by alleviating oxidative stress and mitochondrial dysfunction and reducing AChE activity.

Enhancing Astronaut Resilience: The Role of Elevated ROS in Adapting to Space Radiation

Enhancing Astronaut Resilience: The Role of Elevated ROS in Adapting to Space Radiation

Platinum drugs upregulate CXCR4 and PD-L1 expression via ROS-dependent pathways, with implications for novel combined treatment in gastric cancer.

CXC chemokine receptor 4 (CXCR4) and programmed cell death-ligand 1 (PD-L1) are two critical molecules involved in the tumor immune microenvironment. However, the impact of platinum drugs, such as cisplatin, on CXCR4 or PD-L1 expression and the underlying mechanisms in gastric cancer (GC) remain unknown. Moreover, the correlation between their expression levels in GC remains elusive. Immunohistochemistry, western blot, and RT-qPCR were performed to determine the expression pattern of CXCR4 and PD-L1 in GC. Clinical samples, patient-derived xenografts, and cell-derived xenografts were utilized to investigate the effects of platinum drugs on the expression levels of CXCR4 and PD-L1. Postchemotherapy resected GC tumor tissues showed higher CXCR4 and PD-L1 expression levels than pretreatment biopsies (p < 0.05). Similarly, GC xenografts treated with platinum-based chemotherapy exhibited increased CXCR4 and PD-L1 expression levels compared to saline-treated controls (p < 0.05). A positive correlation was detected between the expression levels of CXCR4 and PD-L1 in GC tumor tissues. Increased levels of CXCR4 and PD-L1 expression, in a dose- and time-dependent manner upon cisplatin treatment, were observed in GC cells (p < 0.05). Cisplatin-induced CXCR4 upregulation relies on ROS/HIF-1α and ROS/NF-κB pathways, while cisplatin-induced PD-L1 upregulation is cyclic GMP-AMP synthase/stimulator of IFN genes-dependent and associated with elevated ROS levels in GC cells. CXCR4 expression was found to be positively correlated with PD-L1 expression in GC. Platinum drugs upregulated the levels of CXCR4 and PD-L1 expression in GC. A combined strategy targeting CXCR-4 and PD-L1 might have clinical prospects for GC patients.

Apoptotic and Molecular Mechanisms of Carthamidin in Breast Cancer Therapy: An Integrated In Vitro and In Silico Study.

The current study examines the anticancer properties of the chemical carthamidin in breast cancer through in-vitro and in silico analysis. This study's results demonstrated that carthamidin strongly inhibited the proliferation of MCF 7 cells in vitro, as evidenced by an IC50 value of 128.65 µg/mL at 24 h, determined using the MTT test. Laser confocal microscopy utilizing AO/EB labeling validated apoptotic effects through upregulating pro-apoptotic cell markers. At the same time, the ROS assay demonstrated elevated ROS production in the treated cells. LDH leakage was corroborated by leakage analysis, revealing high LDH levels at 100µg/mL. The cellular growth parameters were subsequently examined via flow cytometry, showing that the cell cycle was halted in the G0/G1 phase, with 82.9% of the cells residing there. The molecular docking research demonstrated that carthamidin displayed a significant binding affinity with Notch receptors - NOTCH 1-4 and p53, with binding scores ranging from - 5.027 to - 7.402kcal/mol. The results suggest that carthamidin has therapeutic potential in inducing apoptosis and impairing cancer cells, warranting further investigation in breast cancer treatments.

Adipose mesenchymal stem cell-derived extracellular vesicles regulate PINK1/parkin-mediated mitophagy to repair high glucose-induced dermal fibroblast senescence and promote wound healing in rats with diabetic foot ulcer.

Diabetic foot ulcers (DFUs) cause prominent morbidity and mortality. Adipose mesenchymal stem cell (ASC)-derived extracellular vesicles (EVs) show property in facilitating diabetic wound healing, and we explored their role in DFU rats. ASCs were cultured in vitro, passaged and then identified by flow cytometry and induction of osteogenic/adipogenic differentiation. ASC-EVs were extracted and identified. DFU rat model was treated with ASC-EVs. High glucose (HG)-induced rat dermal fibroblasts were treated with ASC-EVs or 3-MA and sh-PINK1 plasmid in vitro. Wound healing was observed. Histological changes, inflammatory cytokines (TNF-α, IL-1β), and α-SMA and p21 double-positive cell level were assessed by HE staining, ELISA, and immunofluorescence. Mitochondrial membrane potential (MMP), cell viability and senescence, and ROS production in cells were assessed by fluorescence dye JC-1, CCK-8, SA-β-gal staining, and ROS kit. p21, LC3II/I, p62, PINK1 and parkin protein levels were determined by Western blot. DFU rats had slow wound healing and elevated levels of IL-1β, TNF-α, α-SMA and p21 double-positive cells, and SA-β-gal, while HG-induced cells had weakened viability, elevated ROS, SA-β-gal, p21 and p62 protein levels, and decreased LC3II/I, PINK1 and parkin protein levels and MMP, which were reversed by ASC-EVs. HG inhibited mitophagy by suppressing the PINK1/parkin pathway to accelerate dermal fibroblast senescence. The PINK1/parkin pathway inhibition partly mitigated the effect of ASC-EVs. ASC-EVs promoted mitophagy by activating the PINK1/parkin pathway in vivo. ASC-EVs mediated mitophagy by activating the PINK1/parkin pathway, thereby impeding HG-induced rat dermal fibroblast senescence and promoting wound healing in DFU rats.

Preparation and Antibacterial Activity Evaluation of Daphnetin-Loaded Poloxamers/Polyvinylpyrrolidone Thermosensitive Hydrogels.

Antibiotic misuse and bacterial resistance are pressing issues threatening public health. Natural plant extracts with bactericidal properties offer potential alternatives to reduce or replace antibiotic use. This study aims to develop a thermosensitive hydrogel containing daphnetin (DAP-TG) using poloxamers 407 (P407), polyvinylpyrrolidone (PVP), and poloxamers 188 (P188). We systematically evaluated the gel's antibacterial activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus), as well as its antibacterial mechanisms. By examining the gelation temperature and time, degradation time, and in vitro release performance of DAP-TG, we produced a sustained-release DAP-TG with a rapid phase transition at (31.6 ± 0.1) °C. Its structure was characterized through Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM). The results indicated that the DAP thermosensitive hydrogel was formed and presented a 3D network spatial structure. The biocompatibility of DAP-TG was explored through the hemolysis test and cytotoxicity test. The results indicated that DAP-TG possessed excellent biocompatibility. The antibacterial efficacy of DAP-TG against E. coli and S. aureus was assessed using minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), growth curve, and inhibition zone tests. Results showed that DAP-TG exhibited excellent antibacterial activity against both E. coli and S. aureus, with MIC values of 1.28 and 0.32 mg/mL. The antibacterial mechanism of DAP-TG was preliminarily explored through the investigation of bacterial cell content leakage, AKP leakage, membrane permeability, SEM, ROS production, and biofilm inhibition activity. DAP-TG induced irreversible damage to the cell membranes of E. coli and S. aureus, resulting in enhanced permeability, elevated ROS levels, and inhibited biofilm formation. Our study indicates that DAP-TG exhibits effective sustained-release and antibacterial properties against E. coli and S. aureus in vitro, making it a promising candidate for antibacterial applications in food and pharmaceutical products.

A Lucknolide Derivative Induces Mitochondrial ROS-Mediated G2/M Arrest and Apoptotic Cell Death in B16F10 Mouse Melanoma Cells.

Melanoma is an aggressive skin cancer with a high risk of cancer-related deaths, and inducing apoptosis in melanoma cells is a promising therapeutic strategy. This study investigates the anti-tumor potential of a novel lucknolide derivative LA-UC as a therapeutic candidate for melanoma. Lucknolide A (LA), a tricyclic ketal-lactone metabolite isolated from marine-derived Streptomyces sp., was chemically modified by introducing a 10-undecenoyl group to synthesize LA-UC. LA-UC preferentially inhibited the proliferation of melanoma cells, including B16F10, while exerting minimal effects on normal melanocytes or other tumor cell types, indicating the selective action of LA-UC against melanoma cells. LA-UC decreased G2/M checkpoint proteins, including cyclin B1 and Cdc2, while activating caspase-3 and caspase-9, resulting in G2/M cell cycle arrest and inducing apoptotic cell death in B16F10 cells. The addition of a pan-caspase inhibitor confirmed the caspase-dependent mechanism of LA-UC-induced cell death. Additionally, LA-UC elevated mitochondrial ROS levels, leading to mitochondrial membrane disruption, upregulation of pro-apoptotic proteins, and DNA damage in melanoma cells. The ROS scavenger N-acetylcysteine reduced LA-UC-induced mitochondrial ROS accumulation, mitochondrial membrane disruption, DNA damage, and apoptosis. Collectively, these findings suggest that LA-UC induces G2/M cell cycle arrest and caspase-dependent apoptosis in B16F10 cells through excessive mitochondrial ROS generation, membrane impairment, and DNA damage, highlighting its potential as a promising therapeutic candidate for melanoma treatment.

Syzygium aromaticum Extract Mitigates Doxorubicin-Induced Hepatotoxicity in Male Rats.

Doxorubicin (DOX), an anticancer drug, is used to treat several types of tumors, but it has detrimental side effects that restrict its therapeutic efficacy. One is the iron-dependent form of ferroptosis, which is characterized by elevated ROS production and iron overload. Syzygium aromaticum has a diverse range of biological and pharmaceutical actions due to their antioxidant properties. This study investigated the effect of S. aromaticum extract (SAE) on hepatotoxicity caused by DOX in rats. Phytochemical analysis was performed to assess compounds in SAE. The ADMETlab 2.0 web server was used to predict the pharmacokinetic properties of the most active components of SAE when DOX was injected into rats. Molecular docking studies were performed using AutoDock Vina. Forty male Sprague Dawley rats were divided into four groups of ten rats each (G1 was a negative control group, G2 was given 1/10 of SAE LD50 by oral gavage (340 mg/kg), G3 was given 4 mg/kg of DOX intraperitoneally (i.p.) once a week for a month, and G4 was administered DOX as in G3 and SAE as in G2). After a month, biochemical and histopathological investigations were performed. Rats given SAE had promising levels of phytochemicals, which could significantly ameliorate DOX-induced hepatotoxicity by restoring biochemical alterations, mitigating ferroptosis, and upregulating the NRF-2-SLC7A-11-GPX-4 signaling pathway. These findings suggest that SAE could potentially alleviate DOX-induced hepatotoxicity in rats.

Guanidinium-Functionalized Carbon Dots: An Efficient Antibacterial Agent against Multidrug-Resistant ESKAPE Pathogens.

The rise of multidrug-resistant (MDR) bacteria poses a substantial challenge in clinical settings, particularly with the increasing prevalence of ESKAPE pathogens (E. faecium, S. aureus, K. pneumoniae, A. baumannii, P. aeruginosa, and E. coli) as critical MDR bacteria. These ESKAPE pathogens have the capability to undermine antibiotic treatments, leading to a high incidence of drug resistance. However, the development of efficient antibacterial agents against ESKAPE pathogens is still in the bottleneck. Herein, the first example of antibacterial carbon dots against ESKAPE pathogens was reported. Onion powder-based carbon dots were melted with poly(hexamethylene biguanide) hydrochloride (PHMB) to obtain guanidinium-functionalized carbon dots (GCDs), which exhibited satisfactory antibacterial activity against all the tested bacteria, including both Gram-positive and Gram-negative bacteria, and even ESKAPE pathogens. The efficient antibacterial ability of GCDs derives from the rupture of the bacterial cell membrane and elevated ROS levels. Safety assessments revealed that GCDs neither trigger detectable drug resistance nor exhibit any cytotoxic effects. Furthermore, GCDs effectively promoted wound healing without observable adverse reactions of mixed MDR bacteria-infected wounds in rats. The GCDs also showed excellent long-term stability. These findings indicate that GCDs hold promise as an efficient antibacterial agent for the treatment of MDR strain-caused clinical infected-wound healing.