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Bee bread has numerous nutritional benefits and bioactive compounds. Other bee byproducts have been used as extender additives to improve semen cryopreservation. Here, we examined the effects of supplementing egg yolk extender (EYE) or soybean lecithin extender (SBLE) with bee bread extract (BBE) on the quality of cryopreserved ram semen. Semen was collected from five adult Rahmani rams once a week for 7 weeks. EYE and SBLE were supplemented with BBE. Antioxidant capacity and total phenolic compound, total flavonoid compound, and total soluble carbohydrate levels of BBE were measured. Sperm characteristics, including progressive motility, viability, abnormalities, membrane integrity, and acrosome integrity, were analyzed after equilibration, thawing, and thawing followed by a 2-h incubation. The total antioxidant capacity and malondialdehyde, hydrogen peroxide, aspartate transaminase, alanine transaminase, alkaline phosphatase, and total acid phosphatase levels in extenders were determined after thawing. Sperm apoptosis was analyzed using annexin V assays. SBLE was more effective than EYE for cryopreserving ram semen. Extender supplementation with BBE improved ram semen quality during freezing in a concentration-dependent pattern. Motility, vitality, and membrane integrity were particularly enhanced in BBE-treated semen. Additionally, BBE promoted antioxidant and enzymatic activities and reduced apoptosis in semen. Thus, extender supplementation with BBE improved sperm cryopreservation.

Bee pollen (BP) is a beehive product known for its therapeutic properties. In this study, we aimed to evaluate 38 BP samples harvested from January 2022 to September 2022 at apiaries located in 6 provinces of Iran. The botanical origin of BP samples was determined using Scanning Electron Microscopy (SEM). The total phenolic content (TPC) of BP samples was evaluated using the Folin_Ciocalteu method. The total aerobic mesophiles were observed using Plate Count Agar (PCA). Based on the predominant grains, most samples were classified as monofloral, identifying 23 plant families. The results indicated a direct relationship between the increased abundance of plant families Asteraceae, Brassicaceae, Rosaceae, and Caryophyllaceae and the increased amount of TPC in BP samples, with the most significant positive influence of the Brassicaceae plant family. The mean value of TPC of methanol extracts of BP samples was 18.48 ± 1.97 mg of gallic acid equivalents per g (GAE/g). The mean value of the total aerobic mesophiles was 23,668.12 colony-forming units per g (CFU/g). The results were based on hygiene standards for human consumption. The current study is considered the first step toward standardizing Iranian BP.

Bee pollen, a natural product rich in polyphenols, exhibits remarkable antioxidant, anti-inflammatory, and hepatoprotective properties. This study was aimed at evaluating the hepatoprotective effects of solid lipid nanoparticles (SLNs) loaded with bee pollen. SLNs were formulated and optimized by varying surfactant ratios and lipid contents at two different temperatures. The optimized bee pollen SLNs demonstrated a particle size of 118.6 nm, a PdI of 0.35, a zeta potential of -22.6 mV, and an entrapment efficiency of 92.7%. The in vitro release study showed minimal release during the initial 120 min, followed by a continuous increase up to 48 h, indicating a sustained and prolonged release profile. Both bee pollen and bee pollen ethanolic extract-loaded SLNs exhibited significant cytoprotective effects against lead-induced cytotoxicity in HepG2 cells. In vivo studies revealed that treatment with bee pollen and especially bee pollen SLNs substantially ameliorated lead-induced hepatic injury. Treatment notably reduced serum levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, lactate dehydrogenase, malondialdehyde, and nitric oxide. Additionally, it enhanced the activity of glutathione peroxidase, catalase, superoxide dismutase, and total antioxidant capacity, as well as levels of total thiol, reduced glutathione, and liver tissue proteins. Histopathological analysis further confirmed that treatment, particularly with bee pollen SLNs, significantly improved lead-induced hepatic structural damage. These findings confirm that bee pollen, and more prominently its SLN formulation, possesses strong hepatoprotective potential.

Bee pollen possesses favorable anticancer activities. As a medicinal plant source, bee pollen (SCBP) possesses potential pharmacological properties, such as reducing cisplatin-induced liver injury, but its anti-liver cancer effect is still rarely reported. This paper aims to investigate the effect and mechanism of SCBP extract (SCBPE) on hepatocellular carcinoma HepG2 cells. The effect of SCBPE on cell proliferation and migration of HepG2 cells was evaluated based on MTT assay, morphology observation, or scratching assay. Furthermore, tandem mass tag-based quantitative proteomics was used to study the effect mechanisms. The mRNA expression levels of identified proteins were verified by RT-qPCR. Tandem mass tag-based quantitative proteomics showed that 61 differentially expressed proteins were obtained in the SCBPE group compared with the negative-control group: 18 significantly downregulated and 43 significantly upregulated proteins. Bioinformatic analysis showed the significantly enriched KEGG pathways were predominantly ferroptosis-, Wnt-, and hepatocellular carcinoma-signaling ones. Protein-protein interaction network analysis and RT-qPCR validation revealed SCBPE also downregulated the focal adhesion-signaling pathway, which is abrogated by PF-562271, a well-known inhibitor of FAK. This study confirmed SCBPE suppressed the cell proliferation and migration of hepatocellular carcinoma HepG2 cells, mainly through modulation of ferroptosis-, Wnt-, hepatocellular carcinoma-, and focal adhesion-signaling pathways, providing scientific data supporting adjuvant treatment of hepatocellular carcinoma using SCBP.

This study investigates the phenolic and sugar composition of an aqueous Syrian Citrus Bee Pollen (SCBP) extract and its application in the green synthesis of gold (AuNPs) and silver nanoparticles (AgNPs). High-performance liquid chromatography (HPLC) identified key phenolic compounds, including caffeic acid, cinnamic acid, kaempferol, apigenin, quercetin, and chrysin. The extract exhibited a high total sugar content (45.8 g/100 g), with total phenolic and flavonoid contents of 240.3 ± 0.04 mg GAE/g and 55.6 ± 0.03 mg QE/g, respectively. Experimental conditions were specifically set as follows: for AuNPs, SCBP extract 1%, HAuCl4·3H2O 2mM, pH 9.5, at room temperature; for AgNPs, SCBP extract 1%, AgNO3 2 mM, pH 10.5, at room temperature. AuNPs Formation was indicated by a purple color and a surface plasmon resonance (SPR) peak at 539 nm. Dynamic light scattering (DLS) analysis revealed a hydrodynamic size of 54.28 nm, a polydispersity index (PDI) of 0.144, and a zeta-potential (ζ) of − 26.9 mV. Field emission scanning electron microscopy (FESEM) showed spherical AuNPs with an average size of 36 nm. AgNPs showed a dark brown color with an SPR peak at 409 nm. DLS measurements indicated an average size of 54.33 nm, a PDI of 0.241, and a ζ-potential of − 25.5 mV. FESEM confirmed nearly spherical AgNPs averaging 49.9 nm with noticeable aggregation. Both nanoparticles showed promising antibacterial activity against S. aureus and S. epidermidis, with inhibition zones measuring 20 mm and 16 mm for AuNPs, and 13 mm and 11 mm for AgNPs, respectively. These findings demonstrate the potential of SCBP-mediated metal nanoparticles as eco-friendly, biocompatible antibacterial agents suitable for biomedical applications such as wound dressing and surface coating.

Bee pollen possesses an anti-cardiomyocyte injury effect by reducing oxidative stress levels and inhibiting inflammatory response and apoptosis, but the possible effect mechanism has rarely been reported. This paper explores the effect of the extract of lotus bee pollen (LBPE) on cardiomyocyte hypertrophy (CH) and its mechanism. The main components of LBPE were identified via UPLC-QTOF MS. An isoproterenol-induced rat H9c2 CH model was subsequently used to evaluate the protection of LBPE on cells. LBPE (100, 250 and 500 μg∙mL−1) reduced the surface area, total protein content and MDA content, and increased SOD activity and GSH content in CH model in a dose-dependent manner. Meanwhile, quantitative real-time PCR trials confirmed that LBPE reduced the gene expression levels of CH markers, pro-inflammatory cytokines and pro-apoptosis factors, and increased the Bcl-2 mRNA expression and Bcl-2/Bax ratio in a dose-dependent manner. Furthermore, target fishing, bioinformatics analysis and molecular docking suggested JAK2 could be a pivotal target protein for the main active ingredients in the LBPE against CH. Ultimately, Western blot (WB) trials confirmed that LBPE can dose-dependently inhibit the phosphorylation of JAK2 and STAT3. The results show that LBPE can protect against ISO-induced CH, possibly via targeting the JAK2/STAT3 pathway, also suggesting that LBPE may be a promising candidate against CH.

Bee pollen is a natural product with multifunctional properties, containing abundant bioactive compounds, especially phenolic acids and flavonoids, which are largely responsible for its antioxidant and antimicrobial activities. In this study, the bioactive composition, antioxidant capacity, encapsulation efficiency, antimicrobial activity, and gastrointestinal stability of bee pollen extract (PE) were investigated. The pollen extract exhibited high total phenolic (2817 mg GAE/100 g) and flavonoid contents (5255 mg QE/100 g), along with strong antioxidant activity (DPPH: 4305 mg TE/100 g; CUPRAC: 3685 mg TE/100 g). To improve the stability and bioaccessibility of phenolic compounds, PE was encapsulated using β-cyclodextrin (BCD) at different weight ratios. Among the formulations, the PE:BCD ratio of 1:2 showed the highest encapsulation efficiency (64%) and favorable physicochemical properties, including higher particle size and more negative zeta potential values, indicating good colloidal stability. Antimicrobial activity was evaluated for PE, BCD-only, and the selected PE-loaded formulation (1:2, w:w). Encapsulation led to a modest reduction in antimicrobial activity compared to free PE (6.25–50 mg/mL); however, the encapsulated formulation still exhibited considerable antibacterial effects against both Gram-positive and Gram-negative strains (25–50 mg/mL). Furthermore, in vitro gastrointestinal digestion indicated that BCD encapsulation substantially enhanced the bioaccessibility of total phenolics (81%) and antioxidant capacity (DPPH: 48%; CUPRAC: 76%), particularly during the intestinal stage. Phenolic profiling showed that chlorogenic acid and quercetin derivatives remained relatively stable throughout digestion. Overall, encapsulation with BCD effectively safeguarded pollen phenolics, improved their gastrointestinal stability, and increased bioaccessibility, highlighting the potential of encapsulated bee pollen as a functional food ingredient or nutraceutical.