Background: Male infertility is increasingly associated with environmental and occupational exposure to organophosphate pesticides, particularly chlorpyrifos (CPF). Chlorpyrifos persists in food, water, and the environment, resulting in chronic low-dose exposure. Although the toxic effects of high-dose exposure are well documented, the reproductive consequences of subchronic low-dose exposure remain unclear. Objectives: To evaluate whether intraperitoneal administration of naringin attenuates CPF-induced reproductive toxicity in adult male rats by assessing sperm parameters, serum reproductive hormones, oxidative stress markers, and testicular histopathology. Methods: Adult male rats were assigned to six groups (n = 5 per group): Control, naringin control (200 mg/kg), CPF (8 mg/kg), and CPF plus naringin (50, 100, or 200 mg/kg). Sperm count, motility, and morphology; serum testosterone, luteinizing hormone, and follicle-stimulating hormone levels; testicular malondialdehyde, reactive oxygen species, glutathione, and superoxide dismutase levels; and histopathological changes were assessed. Results: Chlorpyrifos exposure was associated with impaired sperm count, motility, and morphology; reduced reproductive hormone levels; increased oxidative stress; and testicular histological damage. Naringin pretreatment was associated with dose-dependent improvements in sperm parameters, partial restoration of hormonal balance, improvement in oxidative stress markers, and preservation of testicular histology, with more pronounced effects at 100 and 200 mg/kg. Conclusions: Naringin administration was associated with attenuation of CPF-induced reproductive alterations and improved oxidative stress markers and hormonal parameters. These findings support further investigation of naringin for pesticide-induced male reproductive dysfunction; however, validation in humans is required. The limited range of naringin doses and the absence of molecular investigations highlight the need for further studies to elucidate the underlying mechanisms and translational relevance to human reproductive health.
Background: Bone-associated disorders, including metastatic bone lesions, osteoporosis, and pathological fractures, require agents that can support both diagnostic imaging and targeted therapy. Theranostic radiopharmaceuticals provide an integrated approach for disease detection and radionuclide therapy. Objectives: This study aimed to design, synthesize, radiolabel, and evaluate a novel bone-targeting theranostic radiopharmaceutical, 188Re-MAX-HEDP, for potential diagnostic and therapeutic applications for bone-related diseases. Methods: A novel ligand, MAX-HEDP, was synthesized by conjugating hydroxyethylidene diphosphonic acid (HEDP) to the chelating moiety methoxy amido xanthate (MAX). The ligand was radiolabeled with rhenium-188 (188Re). Labeling efficiency, chemical yield, in vitro stability under physiological conditions, and biodistribution in normal mice were evaluated. Results: The synthesis of MAX-HEDP achieved an overall chemical yield of 91%. Radiolabeling with 188Re produced a highly stable complex, with a labeling efficiency exceeding 99%. Stability studies showed that more than 96% of the complex remained intact at ambient temperature, whereas degradation at 37 °C was less than 10%. Biodistribution studies demonstrated pronounced skeletal uptake of 188Re-MAX-HEDP compared with uptake in other organs, confirming its strong bone-targeting potential. Conclusions: 188Re-MAX-HEDP demonstrated a high synthesis yield, excellent radiolabeling efficiency, favorable stability, and selective skeletal accumulation. These findings suggest that it is a promising, cost-effective theranostic candidate for further clinical evaluation in the diagnosis and radionuclide therapy of bone malignancies and other bone-associated disorders.
Background:The liver, the body's central organ for metabolism and detoxification, is continually exposed to environmental and dietary toxins such as acrylamide (ACR). ACR is commonly found in roasted and fried foods. Strong scientific evidence indicates that ACR exposure causes serious liver injury. Black soldier fly larvae (BSFL) are an environmentally friendly insect, and their extract is rich in valuable bioactive compounds with antioxidant properties. Objectives:This study aimed to investigate the protective effects of the n-hexane extract of black soldier fly larvae against oxidative stress, inflammation, and histopathological changes associated with ACR-induced hepatotoxicity in rats. Methods:Thirty-five male Wistar rats weighing 200 to 250 g were randomly assigned to five equal groups: control, ACR (20 mg/kg), BSFL (360 mg/kg), BSFL (180 mg/kg) + ACR, and BSFL (360 mg/kg) + ACR. At the end of treatment on the twenty-eighth day, the animals were euthanized, and samples were collected for assessment of liver enzymes, oxidative stress, inflammation, ER stress, apoptosis markers, histopathology, and endoplasmic reticulum stress-related protein expression. Data were analyzed using one-way ANOVA followed by Tukey's post-hoc test, with P < 0.05 considered statistically significant. Results:ACR administration significantly increased AST, ALT, ALP, MDA, and NO levels (P < 0.001), significantly decreased SOD, GSH, GPx, and CAT levels (P < 0.001), and significantly increased TNF-α, IL-1β, and IL-6 levels (P < 0.001) compared with the control group. These ACR-induced biochemical and inflammatory abnormalities were confirmed by histopathological observations of liver tissue. Administration of BSFL extract, at the more potent dose of 360 mg/kg, significantly reversed these biochemical, inflammatory, and hepatic markers. BSFL extract was associated with changes in ER stress-related proteins, apoptosis markers, NF-κB, and MAPK family members, suggesting a potential role in GRP78-mediated ER stress signaling. Histological results showed that BSFL extract (360 mg/kg) reduced steatosis, cellular swelling, and lobular inflammation induced by ACR exposure. Conclusions:The results of this study indicate that BSFL extract, a valuable bioactive substance with antioxidant properties, may attenuate alterations in biochemical indices and reduce oxidative stress and inflammation caused by ACR-induced hepatotoxicity.
Background:Fibroblast activation protein (FAP) is a promising molecular target for cancer theranostic applications. However, current fibroblast activation protein inhibitors (FAPIs) have limitations, including rapid clearance and limited tumor retention. Objectives:This study aimed to develop a novel PEG3-linked FAPI derivative, [68Ga]Ga-FAPI-MKG, with enhanced tumor accumulation and retention. FAPI-MKG was prepared by incorporating a PEG3 linker into the FAPI-04 structure to optimize its pharmacokinetic profile. Methods:The compound was synthesized via an 11-step route starting from quinine sulfate and radiolabeled with gallium-68. In vitro studies included determination of lipophilicity (Log P) and stability assays in saline and human serum albumin. Preclinical evaluation in BALB/c mice bearing CT-26 tumors included biodistribution, blocking studies, and PET/CT imaging. Molecular docking and molecular dynamics simulations were performed to provide mechanistic insights into binding interactions. Results:[68Ga]Ga-FAPI-MKG was successfully synthesized with high radiochemical purity (> 98%) and a molar activity of 414.79 mCi/μmol. It demonstrated moderate hydrophilicity (Log P = -3.26 ± 0.18) compared with the reference radiotracer, [68Ga]Ga-FAPI-46 (-3.58 ± 0.29), and high stability (RCP > 90% after 120 minutes). In vivo studies showed significantly higher tumor uptake (7.18 ± 0.56% ID/g at 60 minutes; 3.20 ± 0.11% ID/g at 120 minutes) and prolonged retention compared with the reference radiotracer, along with dual hepatobiliary and renal excretion pathways. Blocking studies confirmed FAP-specific uptake. Computational analyses indicated strong binding energy (-9.8 kcal/mol) and optimized electrostatic interactions with FAP. The strategic incorporation of a PEG3 linker into [68Ga]Ga-FAPI-MKG significantly improved tumor accumulation, extended tumor retention, and increased the tumor-to-background ratio. Conclusions:These findings suggest that [68Ga]Ga-FAPI-MKG may be a promising candidate for clinical translation for imaging and theranostics of FAP-expressing cancers.
Background:Endophytic fungi, which inhabit internal plant tissues without causing apparent disease, have emerged as rich sources of bioactive secondary metabolites. Hypericum species are well-known medicinal plants with diverse pharmacological properties; however, the endophytic fungi associated with these species remain largely unexplored. Objectives:This study aimed to isolate and molecularly identify endophytic fungi associated with Hypericum helianthemoides, H. scabrum, and H. perforatum, and to evaluate the antibacterial activity of their extracts against selected pathogenic bacteria. Methods:Endophytic fungi were isolated from the root and stem tissues of three Hypericum species. Molecular identification was performed by amplifying and sequencing the internal transcribed spacer (ITS) and large subunit (LSU) regions of the rRNA gene. The antibacterial activity of the fungal extracts was assessed by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values against Staphylococcus aureus, S. epidermidis, Bacillus subtilis, Pseudomonas aeruginosa, and Escherichia coli. Results:A total of 20 fungal isolates were obtained, mainly belonging to the phylum Ascomycota, with one isolate (Absidia sp.) belonging to Zygomycota. H. perforatum harbored the highest number of isolates (40%). Most isolates were newly reported endophytes of Hypericum species. The dominant genera were Fusarium and Alternaria. More than half of the isolates exhibited antibacterial activity, particularly against Gram-positive bacteria, whereas only three species were active against Gram-negative strains. Cladosporium subglobosum demonstrated the most potent antibacterial effect, with MIC and MBC values of 3.125 μg/mL. Conclusions:This study highlights the diversity and antimicrobial potential of endophytic fungi associated with Hypericum species. These findings suggest that such endophytes may represent promising sources of novel bioactive metabolites with potential pharmaceutical applications.
Background: Chronic liver injury progressively induces fibrotic remodeling through persistent inflammatory activity and aberrant extracellular matrix accumulation. Growing evidence indicates that this process involves not only intrahepatic signaling but also gut–liver crosstalk and sirtuin-regulated inflammatory pathways. Hesperetin, a citrus-derived flavanone with reported antioxidant and anti-inflammatory properties, has not been adequately evaluated with respect to microbiota-associated changes and SIRT2-associated signaling during fibrotic liver injury. Objectives: This study investigated whether hesperetin attenuates CCl4-induced liver fibrosis in mice by modulating selected gut microbial populations and suppressing SIRT2-associated inflammatory signaling pathways. Methods: Male mice were randomly assigned to the vehicle, CCl4, and CCl4 + hesperetin groups (n = 5 per group). Liver fibrosis was induced by intraperitoneal administration of CCl4 twice weekly for six weeks, and hesperetin was administered orally at 100 mg/kg/day. Histological, biochemical, inflammatory, SIRT2-associated, and selected gut microbial endpoints were assessed. The expression levels of α-SMA, TGF-β1, TNF-α, IL-6, and SIRT2 were quantitatively analyzed using quantitative polymerase chain reaction (qPCR), Western blotting, and an enzyme-linked immunosorbent assay (ELISA). Levels of selected gut microflora were measured using targeted qPCR for Firmicutes, Bacteroidetes, Akkermansia, and Escherichia coli. Results: CCl4 exposure increased collagen deposition, collagen proportionate area (CPA), and serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, indicating fibrotic remodeling and hepatocellular injury. Hesperetin treatment significantly decreased collagen deposition and CPA and partially normalized ALT and AST activities compared with CCl4 treatment alone. Fibrosis biomarkers, including α-SMA and TGF-β1, and inflammatory cytokines, including TNF-α and IL-6, showed significant decreases in expression at both the mRNA and protein levels. Hesperetin reduced CCl4-associated SIRT2 upregulation and restored acetyl-α-tubulin levels, suggesting attenuation of SIRT2-associated deacetylase activity. Conclusions: These exploratory findings suggest that hesperetin attenuates CCl4-induced liver fibrosis by reducing collagen accumulation and biochemical liver injury, suppressing inflammatory and fibrogenic signaling, modulating selected gut microbial populations, and regulating SIRT2-associated acetylation signaling.
Background: Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS) characterized by damage to the myelin sheath and, over time, by progressive neurological disability. Statins have attracted attention in MS research owing to their anti-inflammatory and neuroprotective effects. Objectives: This study assessed whether atorvastatin could ameliorate cuprizone-induced behavioral and histopathological changes in the brains of male C57BL/6 mice. Methods: All groups of mice, except the control group, which received a normal diet, were fed 0.2% cuprizone (CPZ) in their daily diet for 6 weeks to induce demyelination. The treatment groups received atorvastatin (1, 2, or 4 mg/kg/day, i.p.) or nano-atorvastatin (2 mg/kg/day, i.p.) during the final 2 weeks of the study. At the end of the study, behavioral tests were conducted, and immunofluorescence assessment of NF-kB p65 and Nrf2 in the corpus callosum was conducted. Results: CPZ caused progressive weight loss by the end of the study compared with the control group; this effect was reversed by treatment with atorvastatin and nano-atorvastatin. All behavioral tests showed reduced motor coordination in the CPZ group (P < 0.001) compared with the control group. Administration of atorvastatin and nano-atorvastatin during the last 2 weeks reversed these motor deficits. Histopathological examination showed significant demyelination in the CPZ group, which was reversed by atorvastatin injections. Furthermore, CPZ significantly decreased Nrf2 levels (P < 0.001) and increased NF-kB p65 levels (P < 0.001) in the corpus callosum; these changes were stabilized in the atorvastatin and nano-atorvastatin groups. Conclusions: Atorvastatin may attenuate CPZ-induced toxicity by reducing demyelination and modulating the NF-kB p65 and Nrf2 signaling pathways.
Background:Hepatocellular carcinoma (HCC) is a lethal liver malignancy associated with substantial morbidity and mortality and remains a major socioeconomic burden worldwide. Given the high risk associated with HCC, novel therapeutic approaches are urgently needed. Angelica sinensis is a traditional Chinese herbal remedy that contains bioactive phytochemicals with promising anticancer properties, making it a potential candidate for evaluation in HCC. Objectives:This study integrated computational network pharmacology, molecular docking, and in vitro cellular assays to elucidate the mechanisms by which A. sinensis exerts anti-HCC effects in Hep-G2 cells. Methods:Phytochemicals were identified using the Traditional Chinese Medicine Systems Pharmacology database and filtered according to drug-likeness and oral bioavailability criteria. Key compounds (β-sitosterol, alpha-cephalin, sitogluside, and stigmasterol) were further analyzed using the SuperPred Target Prediction tool to predict potential targets. HCC-related targets were curated from GeneCards and refined using the GeneCards Inferred Functionality Score. The overlap between compound-related and disease-specific targets was used to construct a protein-protein interaction network using STRING, which was subsequently visualized in Cytoscape to identify hub genes. Molecular docking between stigmasterol and the top 3 hub genes was evaluated using the CB-Dock2 online server, and all preparations were performed in BIOVIA Discovery Studio. In vitro, Hep-G2 cells were treated with varying concentrations (0, 25, 50, 100, and 200 µg/mL) of A. sinensis extract. Cell viability, clonogenic potential, apoptosis induction, and migratory capacity were evaluated using the MTT assay, clonogenic assay, Annexin V/PI staining, and Transwell migration assay, respectively. Western blotting was used to assess the expression of key hub proteins in Hep-G2 cells. Results:In silico analysis identified an initial pool of 126 phytochemicals, which was refined to 4 key compounds yielding 139 unique targets. Intersection with HCC-related targets produced 123 common targets, generating a network comprising 119 nodes and 520 edges. STAT3, NFKB1, and TLR4 were identified as pivotal hub genes. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses of the hub genes demonstrated significant enrichment in HCC-related pathways and biological processes. Molecular docking indicated a strong binding affinity of stigmasterol to STAT3, NFKB1, and TLR4, with binding energies of -7.8, -6.9, and -6.9 kcal/mol, respectively. In vitro assays showed a dose-dependent reduction in Hep-G2 cell viability and colony formation, significant induction of apoptosis, and marked inhibition of migration. Western blotting confirmed significant downregulation of STAT3, NFKB1, and TLR4 expression at higher extract concentrations. Conclusions:This study concluded that A. sinensis herbal extract was predicted to exert potent antiproliferative effects against HCC through the possible modulation of key signaling pathways and the targeting of hub genes, supporting its potential for further therapeutic development.
Background:Metabolic dysfunction-associated steatotic liver disease (MASLD) is a prevalent metabolic condition marked by abnormal lipid buildup within hepatocytes, leading to inflammation and liver injury. Hepatic lipid accumulation can be driven by multiple factors, including high glucose, free fatty acids, and lipotoxic stress. Autophagy, which may be influenced by metabolic regulators such as cyclic adenosine monophosphate (cAMP), AMP-activated protein kinase (AMPK), and sirtuin 1 (SIRT1), is suggested to contribute to the maintenance of hepatic lipid homeostasis. Tehranolide, a sesquiterpene lactone derived from Artemisia diffusa and structurally related to artemisinin, is believed to have hepatoprotective effects similar to artemisinin. This work is the first to assess the impact of tehranolide on lipid accumulation with emphasis on autophagy/AMPK/SIRT1 signaling in steatotic human hepatoma-derived cells (HepG2). Objectives:This investigation was undertaken to evaluate the potential of tehranolide to reduce lipid accumulation in a high-glucose-induced steatotic hepatocyte model, potentially involving autophagy-related signaling pathways such as cAMP, AMPK, and SIRT1. Methods:A high-glucose-induced steatotic model was established in HepG2 cells. After determining the effective concentration of tehranolide by means of the MTT assay, lipid-loaded cells received treatment with tehranolide. The content of intracellular triglycerides (TGs) was determined via Oil Red O staining and commercial kits. The expression of lipid metabolism-related genes [fatty acid synthase (FASN), sterol regulatory element-binding protein 1c (SREBP-1c), and SIRT1] and autophagy markers [light chain 3 (LC3), beclin-1] was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR), while protein levels of LC3-I, LC3-II, AMPK, and phosphorylated AMP-activated protein kinase (p-AMPK) were evaluated by Western blotting. Intracellular cAMP levels, lactate dehydrogenase (LDH) release, and inflammatory cytokines were also quantified using commercial kits. Results:Tehranolide significantly decreased intracellular TG levels, downregulated lipogenic genes (FASN, SREBP-1c), and upregulated the lipolytic gene SIRT1. It increased the expression of autophagy-related markers (beclin-1 and LC3-II). Furthermore, tehranolide increased intracellular cAMP and AMPK phosphorylation, while inhibition of SIRT1 or blockade of autophagy attenuated these effects. In addition, tehranolide reduced glucose-induced cytotoxicity and suppressed pro-inflammatory cytokine production in HepG2 cells. Conclusions:Tehranolide attenuates lipid accumulation and inflammatory responses in high-glucose-induced steatotic HepG2 cells, potentially involving autophagy-related processes, which may be linked to cAMP, AMPK, and SIRT1. These findings suggest that tehranolide may represent a potential modulator of hepatocellular lipid metabolism in glucose-induced steatosis, warranting further validation in more comprehensive in vitro and in vivo models.
Background: Tropomyosin receptor tyrosine kinase A (TrkA) is essential for cancer cell migration and for stabilizing focal adhesions required for attachment to the extracellular matrix. However, resistance to inhibitors targeting the ATP-binding pocket of the TrkA kinase domain complicates treatment. These inhibitors block ATP binding, which is required for TrkA activation, thereby disrupting downstream signaling pathways involved in cell proliferation and survival in cancers characterized by TrkA overexpression or mutation. Objectives: This study aimed to identify natural compounds as potential TrkA inhibitors for anticancer drug development. The identified compounds exhibited higher binding affinity and greater stability than the control compounds. Methods: Virtual screening, molecular dynamics simulations, MM-GBSA free energy calculations, and principal component analysis were performed using the Schrödinger Desmond software to screen a comprehensive library from the NPACT and PhytoHub databases for potential novel TrkA inhibitors. Results: Two promising inhibitors derived from natural compounds, PHUB000399 and NPACT01417, were identified. Toxicity assessments indicated that both compounds had lower toxicity levels (class 5) than Entrectinib, the reference inhibitor (class 4); the molecular weights of PHUB000399, NPACT01417, and Entrectinib were 275.25, 316.35, and 560.64 g/mol, respectively. Docking studies showed that PHUB000399 and NPACT01417 had superior binding affinities, with grid scores of -13.239 and -13.103, respectively, compared with the co-crystal ligand (-12.567) and Entrectinib (-10.996). Molecular dynamics simulations indicated that the TrkA-PHUB000399 and TrkA-NPACT01417 complexes were more stable than the TrkA-Entrectinib complex. The calculated binding free energies for TrkA-PHUB000399 and TrkA-NPACT01417 were -87.74 and -86.09 kcal/mol, respectively, exceeding those of the co-crystal ligand (-84.14 kcal/mol) and Entrectinib (-73.03 kcal/mol). Conclusions: These findings suggest that NPACT01417 and PHUB000399 are promising candidates for targeting cancer cell migration, with binding affinities comparable to those of established inhibitors. This study provides valuable insights for the development of effective anticancer therapies.
Background: This study evaluated the prophylactic efficacy of nanocurcumin in mitigating chemotherapy-induced peripheral neuropathy (CIPN) among patients with non-metastatic breast cancer receiving paclitaxel. Methods: In this randomized, double-blind, placebo-controlled clinical trial, 80 patients were enrolled after completing anthracycline- and paclitaxel-based chemotherapy regimens. Participants were randomly allocated to receive either nanocurcumin 40 mg twice daily or a matched placebo. The severity of CIPN was evaluated using standardized instruments, including the Neuropathy Disability Score (NDS), Neuropathy Symptom Score (NSS), and Michigan Neuropathy Screening Instrument (MNSI). Large-fiber neuropathy was confirmed by nerve conduction studies. Assessments were performed during the fourth and eighth weeks of the study. Results: Sixty patients completed the trial, including 32 in the placebo group and 28 in the nanocurcumin group. Peripheral neuropathy indices, including NDS, NSS, and MNSI, improved significantly in the nanocurcumin group compared with the placebo group; however, no significant between-group difference was observed in the incidence of large-fiber neuropathy. In addition, nanocurcumin reduced symptoms such as fatigue and muscle cramps, based on NSS and MNSI findings. No adverse events leading to discontinuation were reported in either group. Conclusions: These results suggest that nanocurcumin may be a safe and potentially effective intervention for preventing paclitaxel-induced peripheral neuropathy in patients with breast cancer; however, further research is required to corroborate these findings.
Background: Acute kidney injury (AKI) is a critical clinical condition closely associated with mitochondrial dysfunction. Tanshinone IIA has been shown to exert protective effects against AKI; however, the underlying mechanisms remain unclear. Objectives: This study aimed to investigate whether Tanshinone IIA mitigates lipopolysaccharide (LPS)-induced AKI through SIRTI/PINKi-mediated mitophagy. Methods: A mouse model of AKI was established by intraperitoneal injection of LPS. Renal function was assessed by measuring serum creatinine (Scr) and blood urea nitrogen (BUN) levels. Renal histopathological changes were evaluated using hematoxylin and eosin (HE) staining. Mitochondrial function was assessed by measuring adenosine triphosphate (ATP) content, mitochondrial reactive oxygen species (ROS), and mitochondrial membrane potential in renal tissue. The expression of mitophagy-related markers was analyzed by Western blotting. Results: Lipopolysaccharide treatment significantly increased Scr and BUN levels and induced marked renal pathological injury in mice compared with those in the control group. Tanshinone IIA intervention reduced Scr and BUN levels and alleviated renal tissue damage. Mitochondrial function analysis showed that Tanshinone IIA restored the LPS-induced reduction in ATP content, suppressed excessive mitochondrial ROS generation, and stabilized the mitochondrial membrane potential. Mechanistic analyses further demonstrated that Tanshinone IIA upregulated the LPS-induced expression of SIRT1 and PINK1, increased the LC3-II/LC3-I ratio, and promoted p62 degradation. Conclusions: Tanshinone IIA protects against LPS-induced AKI by improving mitochondrial function through the activation of SIRTI/PINKi-mediated mitophagy.
Background: Ziziphus jujuba is a promising medicinal plant with ethnomedicinal claims and constitutes one of the important herbs in Traditional Chinese Medicine, yet it has not been explored for its anticancer effects against breast cancer (BC). Objectives: The current study aimed to use an integrated approach involving network pharmacology, in-silico molecular docking and dynamics simulations, as well as in-vitro experimental validation, to study the anticancer effects of Z. jujuba in BC. Methods: A total of 133 phytochemicals from Z. jujuba were screened using the TCMSP database, followed by toxicity screening (Protox 3.0) and ADMET analysis (ADMET AI). Soxhlet extraction was performed with a 70:30 ethanol:dichloromethane solvent mixture. Compound-target networks were constructed using SuperPred, SwissTargetPrediction, and Cytoscape (CytoHubba plugin, Degree method). Breast cancer targets (GeneCards, Gifts score >= 60%) were intersected with phytochemical targets (Venny 2.0), and a PPI network was generated (STRING, confidence >= 0.700). GO and KEGG analyses used ShinyGO 0.80. Expression analyses of hub genes were conducted using GEPIA2 (BRCA dataset). Molecular docking was performed with CB-Dock2, and MD simulations used iMODS (100 modes, 300 K) and Desmond. Bioassays included MTT, clonogenic, Annexin V/PI, and Western blot analyses on MDA-MB-231 and MCF-10A cells. Results: Five phytochemicals (Spiradine A, Jujubogenin, Malkangunin, Ceanothic Acid, Moupinamide) showed no toxicity risks. The compound-target network identified 305 targets, with 160 intersecting BC targets. The PPI network (160 nodes, 568 edges, P < 1.0e-16) revealed EGFR, HSP90AA1, and STAT3 as hubs. GO/KEGG analyses linked targets to cancer pathways. EGFR, HSP90AA1, and STAT3 showed higher expression in tumors (P < 0.05), with EGFR/STAT3 linked to poorer survival (P < 0.05). Jujubogenin docking yielded binding affinities of -8.7 (EGFR), -7.5 (HSP90AA1), and -8.2 kcal/mol (STAT3). MD simulations confirmed stable EGFR/STAT3 complexes. The extract exhibited selective cytotoxicity, reduced colony formation, induced apoptosis (P < 0.05), and downregulated EGFR (70%), HSP90AA1 (60%), and STAT3 (90%) at 100 & micro;g/mL (P < 0.01). Conclusions: Network pharmacology revealed that Z. jujuba phytochemicals are predicted to show significant effects against BC. Out of these, Jujubogenin exhibited strong and stable interactions with network-predicted hubs. The extract showed selective cytotoxicity, induced apoptosis, inhibited colony formation, and significantly downregulated these hub proteins, validating its promising anticancer potential.
Background: Cannabis sativa has been used since antiquity for medicinal, ceremonial, and agricultural purposes. Today, cannabis, commonly known as marijuana, remains the most widely used recreational drug worldwide. As its use increases, particularly among young adults, reports of adverse effects on vital organs have also increased. This trend underscores the need for a comprehensive assessment of marijuana’s effects on the cardiovascular system. Objectives: This study aimed to investigate the effects of C. sativa extract on the viability, proliferation, and expression of cardiac markers in human cardiomyocytes. Methods: Human cardiomyocytes (HCMs) were cultured and subsequently exposed to varying concentrations of cannabis extract. Cell viability was assessed using the MTT assay, and morphological changes were examined on days 3 and 6. Growth curves and population doubling times were determined over a 6-day period using the trypan blue exclusion method. In addition, the expression levels of cardiac-specific markers, including GATA4, troponin, and creatine kinase, were quantified using real-time polymerase chain reaction on days 1, 3, and 6. Results: No significant changes in cell morphology were observed; however, cell density was consistently higher in the treated group, particularly at early time points. Growth-curve analysis confirmed these findings, demonstrating higher cell numbers in treated cultures throughout the study period, consistent with the population doubling time data (74.79 h in the treated group vs. 80.45 h in the control group, P = 0.005). Furthermore, the treated group exhibited increased expression of cardiac-specific markers compared with the controls, most notably between day 1 and day 3 (GATA4: 7.97 ± 1.06 vs. 1.48 ± 0.64, P < 0.05; troponin: 3.52 ± 0.85 vs. 1.13 ± 0.92, P = 0.05; creatine kinase: 0.001 ± 0.0008 vs. 1.22 ± 0.90, P = 0.05). Conclusions: Cannabis sativa extract increased proliferation and upregulated the expression of cardiac genes in HCMs. These changes may impair normal cardiomyocyte function, which is vital for cardiovascular health. This issue is particularly important given the increasing global consumption of cannabis-derived substances, driven in part by expanding legalization.
Background: In the biopharmaceutical sector, characterized by rapid technological change and intense market competition, external knowledge search (EKS) has become a strategic necessity to overcome the limitations of closed innovation. However, while EKS is widely studied in developed economies, its underlying mechanisms remain underexplored in emerging biopharmaceutical sectors. Objectives: This cross-sectional study examines how EKS affects innovation performance (IP) in Iranian biopharmaceutical firms, testing the mediating role of knowledge integration capability (KIC) and the moderating effect of R&D intensity, while controlling for firm size and age. Methods: Data were collected using a validated questionnaire completed by 92 senior executives (CEOs, R&D, and Business Development managers) from 44 Iranian biopharmaceutical companies. The model was analyzed using partial least squares structural equation modeling (PLS-SEM) in SmartPLS. Results: EKS significantly enhances both IP (β = 0.462, P < 0.001) and KIC (β = 0.239, P < 0.003). KIC positively influences IP (β = 0.437, P < 0.001) and partially mediates the EKS-IP relationship (indirect effect β = 0.104, P = 0.02). R&D intensity showed no significant moderating effect (β = 0.198, P = 0.10). The model explains 40.5% of IP variance (R² = 0.405). Firm size (β = 0.009, P = 0.924) and age (β = 0.030, P = 0.792) had no significant effects. Conclusions: The impact of EKS on IP depends more on a firm’s KIC than on its R&D intensity. This study advances open innovation theory by clarifying how external knowledge is effectively translated into innovation outcomes within an emerging biopharmaceutical context.
Background: Cancer remains one of the most challenging threats to human health and has prompted intensive research in anticancer drug discovery and synthesis. Isocoumarins are natural lactones with several pharmacological activities, including cytotoxic and anticancer effects. Objectives: This study aimed to synthesize novel ethyl 2-aminopyrano[3,2-c]isochromene-3-carboxylate derivatives and evaluate their cytotoxicity against the MCF-7 breast cancer and A549 lung cancer cell lines, in comparison with a normal cell line (MCF-10A human breast cells). Methods: First, 4-hydroxyisocoumarin was prepared. A series of ethyl 2-aminopyrano[3,2-c]isochromene-3-carboxylate derivatives was then synthesized via a one-pot, three-component reaction of 4-hydroxyisocoumarin, ethyl cyanoacetate, and aromatic aldehydes in ethanol under reflux in the presence of triethylamine. The resulting compounds were characterized using standard spectroscopic techniques, including IR, 1H NMR, and 13C NMR, as well as elemental analyses. Finally, the cytotoxicity of the synthesized compounds was evaluated in MCF-7, A549, and MCF-10A cell lines using a colorimetric MTT assay. Results: The compounds were successfully synthesized, and the cytotoxicity assay demonstrated dose-dependent cytotoxic effects in the tested cell lines. Most compounds exhibited moderate or low toxicity, whereas some were non-toxic in these cells. The most cytotoxic compounds were 4g, 4n, and 4m, with IC50 values of 120.77 ± 7.64, 141.43 ± 13.81, and 168.62 ± 3.59 μg/mL against MCF-7 cells, respectively, and 4a, with an IC50 value of approximately 131.12 ± 11.00 μg/mL against A549 cells. Compared with MCF-10A non-cancerous cells, these compounds showed selectivity indices (SIs) of 4.14, 1.64, 2.47, and 3.42, respectively. Conclusions: The compounds were synthesized in high yields and exhibited moderate-to-mild toxicity toward MCF-7, A549, and MCF-10A cells. Notably, halogen substitution at the ortho position of the phenyl ring increased toxicity, particularly in the MCF-7 cell line. However, the lack of selectivity observed for most compounds indicates that further structural refinement is required before this scaffold can be considered a viable anticancer lead.
Background: Building on our previous investigation of the total semipolar extract (SPE) of Artemisia kopetdaghensis in Plasmodium berghei-infected mice, the present study focuses specifically on the methoxylated flavonoid fraction (MFF). Flavonoids are increasingly recognized for their ability to suppress parasite growth and modulate host immunity. To clarify their role, we isolated and characterized the major flavonoid constituents of this plant and evaluated their antimalarial potential, both alone and in combination with chloroquine, in a mouse model. Methods: Aerial parts of A. kopetdaghensis were extracted using a chloroform:acetone (2:1) solvent mixture. The extract was then fractionated by column chromatography. Based primarily on ^1H-NMR spectra, the MFF was selected and further purified by preparative HPLC. Isolated compounds were identified by 1D and 2D NMR and mass spectrometry. In the in vivo antimalarial study, thirty-six infected female Balb/c mice were treated with MFF and evaluated for key parameters. Docking and molecular interaction studies were conducted using AutoDock v4.2.6 software to examine the interactions of constituents with cytokine protein targets: 1D9C [interferon-gamma (IFN-γ)], 1B6C (TGF-β), 1BBN [(interleukin-4 (IL-4)], and 4HR9 (IL-17), separately. Results: Phytochemical analysis of the MFF by HPLC revealed three flavones: 6-methoxytricin (20%), cirsilineol (10%), and cirsimaritin (70%), identified for the first time in A. kopetdaghensis, with cirsimaritin as the dominant constituent. In vivo, MFF treatment significantly reduced parasitemia and enhanced parasite suppression in P. berghei-infected mice. Cytokine profiling demonstrated suppression of TGF-β and IL-4 followed by their recovery, together with marked elevations of IFN-γ and IL-17, indicating balanced modulation between pro-inflammatory and regulatory responses. These immunological findings were corroborated by molecular docking analyses, which confirmed binding interactions of cirsimaritin with cytokine receptor targets, providing mechanistic support for its immunomodulatory activity. Conclusions: This work extends our previous study on the SPE by moving from extract-level observations to constituent-specific insights. By combining phytochemical isolation with in silico receptor interaction analysis, we demonstrate that methoxylated flavonoids, particularly cirsimaritin, are key modulators of host immunity and promising candidates for further development as adjuncts or leads in antimalarial therapy.
Context: Depression and anxiety are debilitating disorders with complex pathophysiologies associated with neurotransmitter deficits, neuroinflammation, and oxidative stress. Apigenin, a dietary flavonoid, has therapeutic potential because of its neuroprotective properties. Evidence Acquisition: We assessed the antidepressant and anxiolytic effects of apigenin in rodent models, focusing on behavioral outcomes, mechanisms involving neuroinflammation, oxidative stress, and neurotransmitter balance, and relevant molecular pathways. A systematic search of PubMed, Scopus, and Embase was conducted through July 2025. In vivo rodent studies evaluating the effects of apigenin on depression or anxiety were included. Exclusion criteria comprised studies addressing other disorders, apigenin derivatives, or non-original research. Data on study design, model, treatment, outcomes, and mechanisms were extracted. Results: Of 953 screened records, 21 studies met the inclusion criteria. Preclinical evidence consistently indicated that apigenin ameliorated depressive- and anxiety-like behaviors across diverse rodent models, frequently demonstrating effects comparable to or greater than those of conventional pharmacological treatments. Mechanistically, apigenin mediated its neuropsychological effects through multiple pathways, including reducing oxidative and nitrosative stress, inhibiting neuroinflammation, modulating monoaminergic neurotransmission, upregulating neurotrophic factors such as brain-derived neurotrophic factor (BDNF) and cAMP response element-binding protein (CREB), and regulating energy metabolism and neurogenesis. Apigenin exhibited a favorable safety profile across all included studies, even at high doses. Conclusions: In rodent models, apigenin exhibits substantial antidepressant and anxiolytic effects through complex mechanisms involving redox modulation, anti-inflammatory activity, neurotransmitter regulation, and neurotrophic support. These findings indicate that apigenin warrants further translational investigation and potential therapeutic development for treating depression and anxiety disorders.
Background:Alpinia purpurata has shown promising effects in alleviating inflammatory conditions; however, its underlying mechanisms require further investigation. Objectives:This study aimed to elucidate the molecular mechanisms underlying the anti-inflammatory activity of A. purpurata rhizome extract (EEAP) and its active metabolites through the TLR4/MyD88 pathway. Methods:The EEAP was analyzed for proximate composition, vitamin C content, total phenolic content (TPC), total flavonoid content (TFC), total monomeric anthocyanin content (TMAC), and metabolite profile, followed by a network pharmacology analysis. Cytotoxicity was evaluated in HEK-293 cells. An in vivo study of carrageenan-induced paw edema in rats was conducted to validate the findings. Results:EEAP contained 42.12% ash, 34.66% moisture, 6.03% protein, 16.60% fat, and 0.59% carbohydrate. The vitamin C content was 941.55 mg/100 g extract, the TPC was 452.9 mg GAE/100 g extract, the TFC was 416.1 mg QUE/100 g extract, and the TMAC was 2770 mg/100 g extract. Functional enrichment analysis identified the TLR4/MyD88 signaling pathway as contributing to IL-23 production. Eight metabolites with anti-inflammatory properties were verified based on their Pa values: glycidyl oleate, 1-stearoylglycerol, alpha-linolenic acid, methyl palmitate, shogaol, 4-methoxybenzaldehyde, ginkgoneolic acid, and curcumene. EEAP was not toxic to HEK-293 cells (IC50= 741.1 μg/mL) compared with quercetin (IC50= 96.73 μg/mL) and cisplatin (IC50= 7.44 μg/mL). EEAP reduced edema volume but did not alter TLR4 or MyD88 expression under the present experimental conditions. Conclusions:EEAP is not toxic to normal cells and exerts an anti-inflammatory effect by reducing edema volume in carrageenan-induced rats; however, it does not alter the TLR4/MyD88 pathway.
Background:Two major challenges frequently encountered during wound healing are excessive inflammatory responses and bacterial infection. Dendrobium officinale polysaccharide (DOP), a major bioactive constituent of Dendrobium officinale, has shown potential for promoting infected wound healing and tissue repair. Objectives:This study evaluated the therapeutic efficacy of DOP in an infected murine wound model, with a specific focus on the role of the SIRT1/HMGB1/NF-κB signaling axis. Methods:Mouse full-thickness skin defect models were established, and animals were divided into four cohorts: control, infected model, DOP-treated, and DOP plus the SIRT1-specific inhibitor EX527. All groups except the control group were inoculated with Staphylococcus aureus. The primary endpoint was wound closure, quantified as the remaining wound surface area on day 14. Secondary outcomes included body weight changes, histopathological injury scores, inflammatory cytokine protein secretion and mRNA transcript levels, SIRT1 expression, HMGB1 subcellular localization, IκBα degradation, and NF-κB p65 phosphorylation. Results:For the primary endpoint, animals receiving DOP had markedly smaller unclosed wound areas on postoperative day 14 than untreated infected animals. Secondary analyses showed that DOP improved body weight recovery and reduced inflammatory cytokine secretion and corresponding mRNA transcript levels. Mechanistically, DOP increased SIRT1 protein levels, promoted HMGB1 deacetylation at the K29 residue with consequent nuclear sequestration, and suppressed IκBα proteolysis and NF-κB p65 phosphorylation. These protective effects were reversed by the SIRT1 inhibitor EX527. Conclusions:By activating SIRT1, DOP promotes K29 deacetylation of HMGB1 and retains this alarmin in the nucleus. The resulting suppression of NF-κB reduces inflammatory injury and counteracts the adverse effects of bacterial infection on wound repair. These findings provide an experimental basis for the potential application of DOP in the management of infected wounds.
