A stability-indicating RP-HPLC method was developed and validated for the simultaneous quantification of five flavonoids, viz., epigallocatechin gallate (EGCG), silibinin (SL), naringenin (NG), genistein (GT), and apigenin (AG), in herbal extracts. Chromatographic separation was achieved on a C18 column using 0.1 M ammonium acetate and methanol at 40:60% v/v. The method demonstrated acceptable resolution, linearity (r 2: 0.9905–0.9969), precision (<2% RSD), recovery (82.09–99.62%), and sensitivity (LOD: 0.068–1.516 μg/mL; LOQ: 0.20–4.59 μg/mL). The method successfully quantified these flavonoids in the aqueous extracts of Phyllanthus emblica L., Brassica oleracea L., Camellia sinensis (L.). Kuntze, Moringa oleifera L., Syzygium cumini (L.) Skeels, and Allium cepa L. Results revealed that EGCG was the major constituent found in all extracts, whilst AG, GT, and SL were found as minor constituents. The method identified six degradation products with a mass balance of >98%. The method demonstrated acceptable green chemistry by the AGREES and GAPI tools.

Sustainable three-dimensional printed devices for thin-film solid-phase microextraction in green and efficient determination of isoflavones in environmental waters.

Rifampicin (RFP) is a well-known first-line antitubercular drug recommended by the World Health Organization. However, RFP-induced hepatotoxicity makes it impractical for use in long-term antitubercular therapy, and its mechanism is still incompletely understood. This study decrypts the mechanism of aberrant cholesterol metabolism in RFP-mediated liver injury and explores the protective effects of genistein (GEN), an isoflavone isolated from the dyer's broom plant (Genista tinctoria), when used in conjunction with RFP. RFP significantly promoted the formation of lipid droplets, increasing cholesterol deposition in liver by reducing cholesterol 25-hydroxylase (CH25H) and its downstream effector LXRα-ABCA to decrease cholesterol efflux. GEN upregulated CH25H and ultimately relieved RFP-induced liver injury; furthermore, replacing endogenous 25-hydroxycholesterol (25HC) both in vivo and in vitro accordingly rescued the hepatotoxicity induced by RFP. Finally, molecular docking and cellular thermal shift assay (CETSA) showed that RFP could bind to CH25H and impair its stability, an interaction that could be reversed by GEN. Thus, RFP restrains cholesterol efflux by reducing the CH25H-LXRα-ABCA axis to trigger steatosis, while GEN restores this pathway by upregulating CH25H, facilitating cholesterol efflux and mitigating RFP-induced liver injury.

Genistein (GEN) exhibits diverse biological activities. This study is conducted to explore the effect of GEN on metabolic dysfunction-associated steatotic liver disease (MASLD) mice. MASLD was induced in mice by a high-fat diet (HFD). The results indicated that GEN significantly reduced HFD-induced body weight and improved lipid homeostasis and glucose tolerance in HFD-fed mice. Furthermore, GEN intervention markedly ameliorated hepatic pathological changes, hepatic dysfunction, and macrophage polarization. Meanwhile, GEN had a beneficial significant impact on colonic pathological changes and the expression of tight junction-related proteins. The results of gut microbiota showed that high-dose GEN intervention was able to down-regulate Firmicutes and Proteobacteria and increase Bacteroidota at the phylum level. At the genus level, GEN intervention increased Lactobacillus, Muribaculum, and Muribaculaceae_unclassified in MASLD mice, and decreased Ruminococcus, Allobaculum, and Helicobacter. In conclusion, GEN attenuated lipid metabolism disorders and hepatic inflammatory reaction via regulating the composition of the gut microbiota and hepatic macrophage polarization in MASLD mice.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-27783-3.

Lung cancer remains a leading cause of cancer-related mortality worldwide, with therapeutic resistance and toxicity limiting the effectiveness of conventional treatments. Natural compounds have attracted significant interest as adjuncts to modern oncology owing to their ability to modulate multiple oncogenic pathways with comparatively low toxicity. Among these, genistein (GEN), piperine (PIP), and resveratrol (RES) represent particularly promising candidates. GEN, a phytoestrogen, exerts antiproliferative and pro-apoptotic effects through phosphoinositide 3-kinase/protein kinase B (PI3K/Akt), mitogen-activated protein kinase (MAPK), and NF-κB (Nuclear Factor kappa B) signaling. PIP, an alkaloid, not only demonstrates cytotoxicity and cell-cycle arrest but also improves the bioavailability of co-administered therapeutics. RES, a polyphenolic compound, regulates AMP-activated protein kinase (AMPK), PI3K/Akt, and MAPK pathways, inhibits angiogenesis and metastasis, and sensitizes cancer cells to chemotherapy. Synergistic combinations of these phytochemicals have shown enhanced apoptotic responses and significantly reduced IC50 values in cancer, underscoring their potential as multitargeted therapeutic agents. Despite encouraging preclinical and early clinical findings, challenges such as poor solubility and bioavailability hinder clinical translation. This review critically explores their synergistic effects while highlighting the challenges in translating these findings into clinical applications for lung cancer. By elucidating the underlying molecular mechanisms and therapeutic synergy, these natural compounds emerge as promising adjuncts for lung cancer therapy, warranting further mechanistic and clinical investigation.

Random-pattern skin flaps are essential in reconstructive surgery but are frequently compromised by ischemic necrosis. Genistein (GST), a soy-derived isoflavone, possesses antioxidant and anti-inflammatory properties and has demonstrated protective effects in various ischemic disorders. However, its role and mechanism in improving flap survival remain unclear. A murine random-pattern skin flap model and bone marrow-derived macrophages (BMDMs) were used. Invivo, flaps were treated with different doses of genistein to determine the optimal concentration and to assess its effects on survival, angiogenesis, oxidative stress, and apoptosis. Invitro, BMDMs were stimulated with LPS and treated with genistein, with or without AMPK (Compound C) or SIRT1 (EX-527) inhibitors, to investigate macrophage polarization and the underlying AMPK/SIRT1 signaling pathway. Genistein administration significantly improved flap survival area, enhanced blood perfusion, promoted angiogenesis, and reduced oxidative stress and apoptosis. Mechanistically, genistein induced a phenotypic shift in macrophages from the pro-inflammatory M1 to the anti-inflammatory M2 type. This effect was mediated by the activation of the AMPK/SIRT1 signaling pathway. Critically, the beneficial effects of genistein on both macrophage polarization and flap survival were abolished upon pharmacological inhibition of AMPK or SIRT1. Genistein enhances the survival of random-pattern skin flaps by reprogramming macrophage polarization from M1 to M2 via the AMPK/SIRT1 signaling pathway. This study reveals a novel molecular mechanism for genistein's protective effect and highlights its potential as a therapeutic strategy to improve outcomes in reconstructive surgery.

Arsenic (As) is a widespread environmental pollutant that poses serious risks to aquatic organisms, particularly affecting neurological health through oxidative damage, endoplasmic reticulum (ER) stress, and apoptosis. Natural bioactive compounds such as genistein (GNT), a soy-derived isoflavone, have shown potential in counteracting heavy metal-induced toxicity due to their antioxidant and anti-apoptotic activities. This study evaluated the neuroprotective effects of dietary GNT (500mg/kg) in Nile tilapia (Oreochromis niloticus) exposed to 10µg/L As for 60days. Arsenic exposure significantly impaired neurobehavioral performance, including reduced ingestive, swimming behaviors, and aggression. Dietary supplementation with GNT effectively ameliorated these behavioral disturbances. At the biochemical level, As exposure decreased the activities of key brain antioxidants, superoxide dismutase, catalase, and reduced glutathione, while increasing malondialdehyde(MDA). Genistein reversed these oxidative imbalances, restoring antioxidant enzyme activities and reducing MDA levels. Furthermore, GNT significantly upregulated brain acetylcholinesterase content and modulated the expression of genes associated with endoplasmic reticulum (ER) stress and apoptosis. Specifically, GNT reversed As-induced dysregulation in the expression of jnk, chop, eif-2a, xbp-1, ire-1a, atf-6, bip, perk, caspase-3, bax, and bcl-2, indicating its role in mitigating ER stress and apoptotic signaling. Histopathological examination confirmed the protective role of GNT against As-induced brain tissue damage. In conclusion, GNT supplementation offers promising neuroprotection against As-induced toxicity via the modulation of oxidative stress, ER stress, and apoptosis in Nile tilapia.

Genistein-loaded nanostructured lipid carriers for intranasal brain delivery: Preparation, optimization, in-vitro evaluation, and amelioration of bioavailability.

Genistein maintains intestinal homeostasis in colitis mice via activating GPR30-Nrf2 signaling pathway.

Polycystic ovary syndrome (PCOS) is a common endocrine disorder marked by hormonal and clinical imbalances. Genistein (GEN), a soy isoflavone with antioxidant properties, has shown promise in PCOS treatment, although its mechanisms remain unclear. This study aimed to investigate the effects of genistein on ovarian dysfunction in a letrozole-induced PCOS mouse model, focusing on steroidogenesis, apoptosis, and PARP-1 signaling. PCOS was induced by oral administration of letrozole (37.5mg/kg.day) for 21days. Mice were then divided into the following three groups (n=10 each) for another 21-day treatment: control (corn oil), PCOS (continued letrozole), and PCOS+GEN (letrozole+genistein, 50mg/kg.day, i.p.). Genistein restored estrous cyclicity in 80% of treated mice versus 0% in the PCOS group (P<0.05). Histologically, it improved follicular morphology, increased granulosa cell thickness and density, and promoted corpora lutea formation. Genistein significantly reduced serum T and P4 concentrations (P<0.05) and modulated expression of steroidogenic proteins (CYP11A1, CYP19A1, STAR). It also decreased cleaved Caspase-3 and cleaved PARP-1 expression (P<0.05), and suppressed abnormal PARylation without affecting total PARP-1 expression levels. Genistein alleviates ovarian dysfunction in PCOS mice by restoring estrous cyclicity, enhancing follicular development, and normalizing hormone concentrations, through regulation of steroidogenic proteins, inhibition of apoptosis, and modulation of PARP-1 activity. These findings support genistein as a potential therapeutic agent for PCOS, targeting the PARP-1-pADPr axis and apoptosis. Further studies are needed to explore upstream mechanisms and evaluate its long-term effects on reproductive health.

Genistein (GT), a soy-derived isoflavone, have received attention due to their possible anticancer effects. The present research is designed to explore the anticancer potential of GT in the therapeutic management of breast cancer (BC) with molecular mechanisms. For this, data have been collected from plausible different online databases, including PubMed, Web of Science, Google Scholar, PubChem, ScienceDirect, Scopus, Springer Link, and Wiley Online. The findings highlight GT’s potential to reduce oxidative damage, induce apoptosis, and modulate the cell lifecycle in BC cells. It inhibits cancer cell proliferation by affecting the human epidermal growth factor receptor 2 (HER2), phosphoinositide 3-kinase/ protein kinase B (PI3K/AKT), and hedgehog regulatory pathways. Moreover, GT exhibits promising pharmacokinetic properties, including high intestinal absorption (95.5%) and strong Caco-2 permeability, with metabolism primarily via CYP1A2 and CYP2C19. However, its poor blood-brain barrier (BBB) permeability and low water solubility present difficulties to the systemic bioavailability. Toxicological investigations demonstrate a dose-dependent effect of GT, with large dosages causing toxicity and low concentrations showing therapeutic potential. Clinical studies emphasize its complex involvement in BC, with differing findings on benefits and risks, indicating the need for additional safety and efficacy study.

Blend of polymeric nanoparticles-in-microneedle arrays: A potential transepidermal route for genistein for anti-melanoma activity.

Abstract Molecular modeling techniques is applied to investigate the mechanisms of α‐Syn aggregation, analyzing the stability of oligomers, as well as evaluating the potential of polyphenols in destabilizing these aggregates. The results indicate that the stability of the aggregates increases with the number of peptide chains, due to the formation of secondary structures in α‐Syn, particularly β‐sheets. Among the models studied, the tetramer is identified as the smallest stable protofibril, making it a target for ligand testing. Regarding the interaction analysis between polyphenols and α‐Syn aggregates, molecular dynamics simulations suggest that quercetin (QC) and hydroxytyrosol (HT), and genistein (GT) interact in the NAC region of the oligomer. Furthermore, the analysis of the total contacts fraction revealed that the presence of the ligand reduces the number of residue‐residue interactions within the oligomer, suggesting a global destabilizing effect on the aggregate structure. In terms of energetic parameters, QC showed the greatest destabilizing effect, followed by GT. HT, in turn, exhibited a localized destabilizing effect. Thus, the present study provides insights into the destabilization of the α‐Syn tetramer by polyphenols, which may contribute to the development of new therapies for Parkinson's disease.

Development and validation of a HPLC-MS/MS methodthe determination of genistein and equol in serum, urine and follicular fluid.

The immune system of fish is severely impacted by heavy metal pollutants that enter the aquatic system, reducing their survival and resistance to diseases. Arsenic is a widespread heavy metal that contaminates aquatic bodies, impacts fish health, and hinders the sustainability of aquaculture. In the current investigation, we studied the arsenic-hazard consequences on the hematology, serum immunological responses, antioxidant activity, autophagy-related genes, splenic architecture, and protection against Aeromonas sobria in Nile tilapia (Oreochromis niloticus). Additionally, we proposed the ameliorative potential of genistein as a feed additive against arsenic exposure. For this purpose, 160 fish were equally set into four groups, each with four replicates for sixty days. The C (control) and GN (genistein) groups were fed basal diets fortified with 0 and 500mg genistein/kg diet and were not exposed to arsenic. However, the AS (arsenic) and AS + GN (arsenic + genistein) groups were reared under arsenic conditions (10µg/L of As2O3) and fed on identical diets as the C and GN groups. The outcomes revealed that arsenic exposure induced hematological disorders and anemic conditions in Nile tilapia. The serum immunological and splenic antioxidant enzymes (phagocytic activity, nitric oxide, lysozymes, complement 3, superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase, glutathione S-reductase, and reduced glutathione) were substantially lowered, and the malondialdehyde level was increased in the arsenic-exposed fish. Arsenic exposure down-regulated the splenic expression of antioxidant enzymes while upregulated the autophagy-related genes (Beclin-1 and AKT-1) and endoplasmic reticulum (ER) stress-related genes (ATF-6, XBP-1, JNK, and BIP). The arsenic-exposed fish showed the lowest survivability (66.7%) when challenged with A. sobria relative to the control (80%). The dietary intervention of genistein in the Nile tilapia diet mitigated the arsenic-induced hematotoxic, immunotoxic, oxidative stress, autophagy, and ER stress consequences. The histological picture of the spleen and the survivability of the arsenic-exposed fish were restored by feeding on the genistein-fortified diet. Genistein is a promising feed additive for counteracting the harmful effects of arsenic in Nile tilapia, which helps in sustainable aquaculture development.

Daidzein (DEIN) and genistein (GEIN) are aromatic amino acid (AAAs) derivatives that have various physiological, nutritional, and medical activities. The low abundance in natural sources and structural complexity limit their acquisition through plant extraction and chemical synthesis. The versatile red yeast Rhodotorula toruloides, endowed with critical enzymes for producing AAAs and derivatives, has been engineered to produce DEIN and GEIN. First, the stability of DEIN and GEIN and the selectivity of chalcone synthase were investigated before constructing the biosynthetic pathway for DEIN and GEIN. Second, ATP-citrate lyase was inactivated to downregulate the lipid biosynthesis and redirect the metabolic flux toward DEIN and GEIN production. Then, the biosynthesis of DEIN and GEIN was enhanced by enzyme fusion. Finally, the production of DEIN and GEIN reached 6.92 and 3.55 mg/L, respectively. Herein, the biosynthesis of DEIN and GEIN was achieved for the first time with R. toruloides, which should provide insights into the bioproduction of AAAs-derived compounds.

BackgroundBreast cancer represents a significant global health concern and is influenced by a range of environmental factors. Increased fat intake and physical inactivity contribute to elevated body fat levels and are strongly linked to breast cancer incidence. Genistein (GEN), isoflavone in soy-derived foods, demonstrates anti-estrogenic properties and anti-cancer effects by regulating various mechanisms such as apoptosis. Regular physical activity prevents the progression and development of cancer by releasing various myokine signaling molecules from the muscles. This study aimed to explore the potential anti-cancer effects of combining GEN supplementation with regular moderate-intensity exercise on breast cancer.MethodsFemale BALB/c mice aged 5 weeks were divided into five groups and received GEN, moderate-intensity exercise, or a combination of both throughout the experiment. After 8 weeks of treatment, mammary tumor cells were inoculated into mammary fat pads. Anti-cancer effects of these treatments on apoptosis, macrophage polarization, and adipose tissue wasting mechanisms in breast tumors were analyzed. In addition, U937, a human monocytic leukemia cell line, was treated with phorbol-12-myristate-13-acetate and interleukin (IL)-4 to induce an M2 macrophage phenotype and analyzed markers for M2 polarization.ResultsModerate-intensity exercise alone or in conjunction with GEN proved effective in retarding tumor initiation and growth, leading to reduced tumor volume compared to GEN supplementation alone. The combined regimen enhanced the expression of apoptosis markers and augmented the proportion of M1 macrophages while diminishing M2 macrophages. In vitro, treatment with GEN and myokines suppressed markers of M2 macrophage polarization and expression of the JAK1/STAT6 signaling pathway. Furthermore, the study suggested that the combined intervention of GEN supplementation and moderate-intensity exercise prevented adipose tissue wasting by regulating adipogenesis, lipolysis, and systemic inflammation in subcutaneous fat.ConclusionsThe potential anti-cancer effects of GEN supplementation and regular moderate-intensity exercise on breast cancer are mediated through the induction of apoptosis and inhibition of macrophage polarization. They also exert a protective effect on adipose tissue wasting.

Breast cancer (BC) is the most prevalent form of cancer among women worldwide, accounting for approximately 36% of cancer cases. Due to its inimitable pathological expression and restricted success of accessible therapeutic modalities, fanatical research in this area is essential. Our group has developed a nanovesicular lipid carrier system consisting of Exemestane (EXM) and Genistein (GNS), which have been successfully incorporated into both uncoated and chitosan-coated liposomes. This combination aims to enhance anticancer efficacy. EXM is known to cause bone loss, while GNS, a natural isoflavone, has been shown in research to possess bone-protective effects. Therefore, we combined these two compounds to mitigate the side effects of EXM. Our previous publication details the formulation development of uncoated EXM-GNS liposomes (EXM-GNS-LPS) and chitosan-coated EXM-GNS liposomes (CH-EXM-GNS-LPS), where we addressed the pharmacotechnical challenges of combining a synthetic drug with herbal drug. Both uncoated and coated liposomes were tested for their budding effects on bone loss induced by hormonal therapy. Pharmacokinetic and pharmacodynamic studies were conducted on rat models with breast cancer, treated with different formulations. Biochemical investigations revealed significant changes in biomarker levels, indicating effects on bone development and resorption. Improvements in bone health and anticancer efficacy were observed to be statistically significant (p < 0.05). Micro-CT analysis of bone samples showed that the chitosan-coated EXM-GNS liposome treatment group yielded the best results when evaluate against other treatment groups. Additionally, histological examination of the bone treated with CH-EXM-GNS-LPS demonstrated a marked restoration of trabecular bone architecture, characterized by a well-connected bone matrix and narrower inter-trabecular spaces compared to the toxic control group. The synergistic effect of EXM and GNS, encapsulated in liposomes, offers an innovative solution to the challenges of breast cancer treatment. The chitosan coating not only improved the stability and controlled release of the drugs but also provided additional benefits in terms of biocompatibility and targeting potential. Overall, the results of this study indicate that the CH-EXM-GNS-LPS formulation holds significant promise as a therapeutic and preventive strategy for bone loss associated with hormonal therapy in breast cancer patients. This work lays the foundation for future clinical applications, highlighting the potential for combining synthetic and natural compounds in advanced drug delivery systems to address complex, multifactorial health issues.

The current study evaluated the effects of genistein (GEN) supplementation to alleviate the arsenic (As)-induced hepatotoxicity in Oreochromis niloticus. This was conducted in two steps: a computational prediction study (in silico) and an experimental investigation (in vivo). The prediction step involved molecular docking analysis to assess the interactions between GEN and key stress-related mRNAs in Nile tilapia. In the experimental phase, 160 Nile tilapia fingerlings were randomly assigned to four treatment groups (in four replicates/group) for 60 days: (1) a control group fed a basal diet, (2) a GEN group receiving a GEN-supplemented diet (500mg/kg), (3) an As group exposed to 10µg/L As, and (4) an As + GEN group, in which fish was exposed to As and fed the GEN-supplemented diet. The computational assessment of GEN's binding ability revealed strong interactions with key mRNAs associated with inflammation and misfolded protein responses. The in vivo results revealed that GEN significantly alleviated As-induced hepatic oxidative stress and hepatocellular damage by restoring liver enzyme levels, lipid profiles, and bilirubin content and restoring the serum proteins to near-normal values. Additionally, GEN downregulated the expression of endoplasmic reticulum (ER) stress- and inflammation-related genes in the liver tissue of the As + GEN group, compared to the As-exposed fish fed on a basal diet. Additionally, the histopathological analysis further confirmed that GEN supplementation mitigated hepatic tissue damage, reducing necrosis, congestion, and inflammatory cell infiltration. In conclusion, GEN supplementation effectively counteracted As-induced hepatotoxicity in Nile tilapia by modulating oxidative stress, ER stress, and inflammation while preserving liver structure and function. Also, the molecular docking results suggest that GEN interacts with the mRNAs of inflammatory and misfolded protein targets, which are increased due to exposure to As-contaminated water. All our findings highlight GEN as a promising natural dietary additive for improving hepatic health in fish inhabiting As-contaminated environments.

Abstract Caviar, a delicacy prepared from sturgeon ovaries, is notoriously expensive. Recent advances have enabled the production of caviar from sturgeons. This study aimed to enhance the production efficiency of female monosex cultures using genistein, an isoflavone with estrogenic effects that is abundant in soybeans. Twenty-five sterlets (Acipenser ruthenus) aged 56 days post-hatch (DPH) were administered one of three feeds containing genistein (GS) for 180 days: GS10 (10 μg GS/g diet), GS100 (100 μg GS/g diet), or GS1000 (1000 μg GS/g diet). A positive control group was fed a diet containing 10 μg 17β-estradiol (E2)/g diet. Following treatment, the fish were reared on commercial feed, and seven to nine sterlets from each group were sampled at 311–312 DPH. A negative control group of ten sterlets raised on a standard diet was sampled. Gonadal morphology was histologically analyzed, the long axis of oocytes calculated, and genetic sex was determined using PCR methodology to assess the effects of genistein administration. The gonads of all genetic females from all of the groups developed into ovarian tissues. In the negative control, GS10, and GS100 groups, genetic male gonads developed into testes. In contrast, genotypic sterlets in the E2 and GS1000 groups developed ovarian tissue, indicating successful feminization via oral genistein administration.