Red Propolis Research Articles

Brazilian red propolis: Chemical composition and antibacterial activity determined using bioguided fractionation

The indiscriminate use of antibiotics is causing an increase in bacterial resistance, complicating therapeutic planning. In this context, natural products have emerged as major providers of bioactive compounds. This work performs a bioguided study of Brazilian red propolis to identify compounds with antibacterial potential and to evaluate their cytotoxicity against non-tumour cells. Using bioguided fractionation performed with the hydroalcoholic extract of red propolis from Alagoas, it was possible to obtain subfractions with remarkable bacteriostatic activity compared with the precursor fractions. The SC2 subfraction was highlighted and showed the best results with minimal inhibitory concentrations (MICs) of 56.75, 28.37, 454.00, and 227.00 μg mL−1 against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa, respectively. However, this study also revealed a cytotoxic effect against the non-tumour Vero cell line. Furthermore, through chemical analyses using high resolution mass spectrometry, high performance liquid chromatography with UV detection, and gas chromatography coupled to mass spectrometry, we verified the presence of important marker compounds in the fractions and extracts, including formononetin (m/z 267.0663), biochanin A (m/z 283.0601), and liquiritigenin (m/z 255.0655). The results obtained in this study suggest an important antibacterial potential of red propolis subfractions. In this context, the bioguided fractionation has been a useful process, due to its ability to isolate and concentrate active compounds in a logical and rational way.

Effect of red propolis on hamster cheek pouch angiogenesis in a new sponge implant model.

To evaluate the effects of red propolis on cheek pouch angiogenesis in a hamster new model sponge implant. Forty eight animals divided into eight groups. (Groups I-IV), the animals were treated for 15 days before and 10 days after sponge implantation. (Groups V-VIII), the animals were treated for 10 days after sponge implantation (GI and GV: red propolis 100 mg/kg, GII and GVI: celecoxib 20 mg/kg, GIII and GVII: 1% gum arabic 5 mL/kg, GIV and GVIII: distilled water 5 mL/kg). On the 11th day of implantation, the animals were anesthetized for stereoscopic microscopic imaging and morphometric quantification of angiogenesis (SQAN), followed by histopathological evaluation (H&E). In the SQAN analysis, no significant difference was found between the groups. However, on histology, propolis was found reduce the population of mastocytes in the qualitative analyses (p = 0,013) in the quantitative analyses to reduce the number of blood vessels (p = 0,007), and increase the macrophage count (p = 0,001). Red propolis inhibited inflammatory angiogenesis when administered before andcontinuously after sponge implant, and was shown to have immunomodulating effects on inflammatory cells (mastocytes and macrophages) in a new sponge implant hamster model.

Development and characterization of microencapsules containing spray dried powder obtained from Brazilian brown, green and red propolis

The aim of this study was to obtain dried powders from the brown, green and red varieties of the Brazilian propolis by spray drying, using gum arabic and maltodextrin as encapsulating materials. Propolis microparticles were evaluated for particle size, X-Ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), moisture, water activity, solubility, hygroscopicity, encapsulation efficiency, total phenolic content, flavonoids and antioxidant activity (DPPH, ABTS+, FRAP, ORAC). Bioactive compounds were determined by UPHLC-QqQ-MS/MS. The results showed that the microparticles presented spherical forms, smooth surfaces, amorphous characteristics, low water activity, moisture and hygroscopicity, high solubility, good encapsulation efficiency, good antioxidant activity, although a reduction in total phenolics and flavonoids from the non-encapsulated propolis. The most important compounds in propolis powders were artepillin C, kaempferide, p-coumaric acid, luteolin, chlorogenic acid, kaempferol and caffeic acid. Spray-drying of propolis is a viable option to increase its use in food and pharmaceutical applications.

Antitrypanosomal, antileishmanial and cytotoxic activities of Brazilian red propolis and plant resin of Dalbergia ecastaphyllum (L) Taub

The treatment for leishmaniasis and Chagas disease can be hard and painful, such that many patients give up on the treatment. In order to find an alternative path for the treatment of these diseases, researchers are using natural products to fight these parasites. The aim of this study was to evaluate the antiprotozoan and cytotoxic activities of red propolis samples collected from different Brazilian states and seasons whilst searching for possible activity differences. We also compared the red propolis results with the ones obtained for the plant resin extract collected from Dalbergia ecastaphyllum trees. The hydroethanolic red propolis extracts from Pernambuco and Alagoas, and the D. ecastaphyllum resin were evaluated regarding their antileishmanial, antitrypanosomal and cytotoxic activity. All extracts showed antiprotozoan and cytotoxic activity. RP-PER showed to be more cytotoxic against protozoan parasites and fibroblast cells. All propolis extracts showed a higher cytotoxic activity when compared to resin extracts. The propolis sample collected in Pernambuco during the rainy season killed the parasites with lower concentrations than the sample collected in the dry season. The IC50 observed against the parasites could be used without high fibroblast cell damage.

Polymeric nanoparticle systems loaded with red propolis extract: a comparative study of the encapsulating systems, PCL-Pluronic versus Eudragit®E100-Pluronic

The aim of the present study was to compare two nanoparticle composition loaded with Brazilian red propolis extract regarding its physicochemical characteristics and its antioxidant and antileishmanial activities. The red propolis nanoparticles in an aqueous medium and in solid-state presented particle size in a nanometric scale with an apparent size of 100–288 nm for the NEPE and 175–380 nm for the NPPE. ATR-FTIR and thermal analysis revealed an encapsulation of flavonoids from the red propolis extract in polymeric matrices for the multidrug delivery system. UPLC-DAD identified red propolis markers (flavonoids) in EPE, NEPE and NPPE. The efficiency of encapsulation (28.0–55.0% for NEPE and 61.2–81.0% for NPPE) were determined and calculated using UPLC-DAD. DPPH method showed the antioxidant activity of both EPE and nanoparticle compositions of red propolis. These polymeric matrices systems were able to encapsulate flavonoids from red propolis extract with specific characteristics of solubility and polarity. EPE and nanoparticles loaded with red propolis extract in the multi-constituent co-delivery system presented leishmanicidal activity and a good correlation was established between IC50 and efficiency of encapsulation. Red propolis nanoparticles exhibited leishmanicidal activity but NEPE presented a lower leishmanicidal effect in relation to NPPE, which showed similar activity compared to EPE. The nanopolymeric matrices choice should be established in propolis nanoparticle compositions to avoid lack of efficacy of bioproducts. Red propolis nanoparticles were shown to be a potential final bulk product for the preparation of various pharmaceutical and cosmetics compositions in therapy against diseases such as leishmaniasis.

Phytochemical screening, antioxidant and antibacterial activities of some commercial extract of propolis

This study investigated the chemical composition, flavonoids, phenolic compounds, antioxidant activity and antibacterial activity of commercial propolis extracts produced in the Sergipe and Alagoas States of Brazil as potential bioproducts for the food and pharmaceutical industries. Four samples were analyzed, three brown propolis extracts and one red propolis extract, and were characterized through phytochemical screening, chemical, chromatographic profile and antibacterial activity. Phytochemical analysis detected the presence of triterpenoids and phenolic compound in propolis extracts. Propolis extracts showed total phenolic content between 9 and 15% and total flavonoids >2%. Propolis extracts showed excellent antioxidant activity with inhibition of the Free radical DPPH˙ between 97 and 60%, which confirm the results obtained in total phenolics, total flavonoids content and antibacterial activity. The chromatographic profile showed differences for brown propolis samples and quite different from the red propolis extract, which present flavonoids as isoflavonoids, pterocarpans, chalcones and guttiferones. Commercial propolis extract (propolis extract C and propolis extract D) showed excellent activity for Staphylococcus aureus ATCC 25923 and moderate activity for Pseudomonas aeruginosa ATCC 27853. The chemical characterization of propolis extracts is fundamental in the process of standardization and monitoring of the chemical composition susceptible to geographic and seasonal variation. These results point to new possibilities of use as bio-preservative of processed foods as well as in the development of pharmaceuticals and nutraceuticals products from propolis extract C and D in its formulation actuate on the inhibition of some pathogenic microorganisms strains.

Mexican Propolis: A Source of Antioxidants and Anti-Inflammatory Compounds, and Isolation of a Novel Chalcone and ε-Caprolactone Derivative.

The propolis produced by bees are used in alternative medicine for treating inflammation, and infections, presumably due to its antioxidant properties. In this context, five propolis from México were investigated to determine their inhibitory lipid peroxidation properties. The ethyl acetate extract from a red propolis from Chiapas State (4-EAEP) was the most potent (IC50 = 1.42 ± 0.07 μg/mL) in the TBARS assay, and selected for further studies. This extract afforded two new compounds, epoxypinocembrin chalcone (6), and an ε-caprolactone derivative (10), as well as pinostrobin (1), izalpinin (2), cinnamic acid (3), pinocembrin (4), kaempherol (5), 3,3-dimethylallyl caffeate in mixture with isopent-3-enyl caffeate (7a + 7b), 3,4-dimethoxycinnamic acid (8), rhamnetin (9) and caffeic acid (11). The HPLC profile, anti-mycobacterial, and antioxidant properties of this extract was also determined. Most of the isolated compounds were also tested by inhibition of reactive oxygen species (ROS) in challenged mouse bone marrow-derived mast cells (BMMCs), and DPPH. Their anti-inflammatory activity was evaluated by TPA, and MPO (myeloperoxidase) activity by ear edema test in mice. The most potent compounds were 7a + 7b in the TBARS assay (IC50 = 0.49 ± 0.06 μM), and 2 which restored the ROS baseline (3.5 μM). Our results indicate that 4-EAEP has anti-oxidant, and anti-inflammatory properties due to its active compounds, suggesting it has anti-allergy and anti-asthma potential.

Chemical composition, immunostimulatory, cytotoxic and antiparasitic activities of the essential oil from Brazilian red propolis.

Most studies of Brazilian red propolis have explored the composition and biological properties of its ethanolic extracts. In this work, we chemically extracted and characterized the essential oil of Brazilian red propolis (EOP) and assessed its adjuvant, antiparasitic and cytotoxic activities. The chemical composition of EOP was analyzed using gas chromatography with mass spectrometry (GC-MS). EOP was tested for in vitro activity against Trichomonas vaginalis (ATCC 30236 isolate); trophozoites were treated with different concentrations of EOP (ranging from 25 to 500 μg/mL) in order to establish the MIC and IC50 values. A cytotoxicity assay was performed in CHO-K1 cells submitted to different EOP concentrations. BALB/c mice were used to test the adjuvant effect of EOP. The animals were divided in 3 groups and inoculated as follows: 0.4 ng/kg BW EOP (G1); 50 μg of rCP40 protein (G2); or a combination of 0.4 ng/kg BW EOP and 50 μg of rCP40 (G3). Total IgG, IgG1 and IgG2a levels were assessed by ELISA. The major constituent compounds of EOP were methyl eugenol (13.1%), (E)-β-farnesene (2.50%), and δ-amorphene (2.3%). Exposure to EOP inhibited the growth of T. vaginalis, with an IC50 value of 100 μg/mL of EOP. An EOP concentration of 500 μg/mL was able to kill 100% of the T. vaginalis trophozoites. The EOP kinetic growth curve showed a 36% decrease in trophozoite growth after a 12 h exposure to 500 μg/mL of EOP, while complete parasite death was induced at 24 h. With regard to CHO-K1 cells, the CC50 was 266 μg/mL, and 92% cytotoxicity was observed after exposure to 500 μg/mL of EOP. Otherwise, a concentration of 200 μg/mL of EOP was able to reduce parasite proliferation by 70% and was not cytotoxic to CHO-K1 cells. As an adjuvant, a synergistic effect was observed when EOP was combined with the rCP40 protein (G3) in comparison to the administration of each component alone (G1 and G2), resulting in higher concentrations of IgG, IgG1 and IgG2a. EOP is constituted by biologically active components with promising antiparasitic and immunostimulatory activities and can be investigated for the formulation of new vaccines or trichomonacidal drugs.

The photoprotective and anti-inflammatory activity of red propolis extract in rats

The negative effects triggered by ultraviolet radiation, such as premature aging and carcinogenesis, have motivated several studies on photoprotection. Recent strategies for photoprotection have included the incorporation of natural antioxidant and anti-inflammatory compounds, such as flavonoids, into sunscreens and the oral administration of natural antioxidant extracts. Brazilian Red propolis extract contains isoflavonoids with antioxidant and anti-inflammatory activities. Here, we investigate the photoprotective effects of orally- or topically-administered formulations containing hydroalcoholic extract of red propolis (HERP) in a rodent model. HERP showed markers identified as: daidzein (4.68 μg/mL), formononetin (31.81 μg/mL) and biochanin A (9.58 μg/mL). A fourth peak was found in the chromatogram but was not identified. The antioxidant activity of HERP was calculated to be 3.07 mmol Trolox/g and 2.13 mmol Trolox/g, respectively. Topical HERP exerted a protective action against UVB radiation, which was similar to that exerted by oxybenzone filter. Oral HERP as an adjuvant treatment did not increase sunburn protection. However, the oral administration of HERP presented chemoprotective and anti-inflammatory activity (p < 0.05) similar or better than Polypodium leucotomos oral treatment (positive control). In conclusion, topical administration of HERP has photoprotective activity in a murine model and the mechanisms of protection can be related to the antioxidant and anti-inflammatory characteristics of HERP compounds.

Evaluation of the antimicrobial activity and cytotoxic effect of hydroxyapatite containing Brazilian propolis

The aim of this work was to produce hydroxyapatite powder (HA) containing the dry extract of green and red propolis, and to evaluate the possible bactericidal activity of these materials over a short period of time through a fast release system. The ethanolic extracts of green and red propolis (EEP) were incorporated into the material by spray drying. After release tests, powders containing dry EEP were characterized regarding the content of total phenolics and flavonoids. Material characterization was undertaken by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). The antimicrobial activity was evaluated by plate colony counting, minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) against Staphylococcus aureus (S. aureus). The cytotoxicity of the materials was determined by the neutral red incorporation method. The materials showed apparently spherical morphology, indicating a decrease in the degree of agglomeration with the addition of propolis. Characteristic HA and propolis functional groups were observed in the FTIR. The materials showed a higher release of phenolics and lower amounts of flavonoids when compared to the EEP, with the higher amounts of flavonoids observed for HA with red propolis. A bactericidal effect was observed for all materials within the interval of 0.5 and 1 h, showing lower inhibitory activity (MIC) and higher bactericidal activity (MBC) when compared to the EEP, with the best results attributed to HA with red propolis. The IC50 values (which is the concentration needed to inhibit cell growth by 50%) obtained from the cytotoxicity assay for HA with the green and red propolis lay between MIC and MCB. Considering these results, it is suggested that HA and propolis may be used as a possible antimicrobial agent, inhibiting the growth of S. aureus, although further in vivo biocompatibility should be investigated before using this material as a medical device with bactericidal potential.

Isoflavonoids from Brazilian red propolis down-regulate the expression of cancer-related target proteins: A pharmacogenomic analysis.

Vestitol and neovestitol are bioactive isoflavonoids isolated from Brazilian red propolis, a unique Apis melifera type of propolis botanically originated from Dalbergia ecastophyllum. Although these molecules have relevant biological effects, including anticancer and immunomodulatory activities, their mechanism(s) of action and the affected pathways remain largely unknown. Here, we carried out a pharmacogenomic analysis to investigate the effects of vestitol and neovestitol on the whole-genome expression in human tumor cells, particularly cancer-related target proteins. HeLa cells were exposed to the compounds at IC20 and genomic information of treated cells was analyzed using the Illumina transcriptome system and GeneGo MetaCore software. Our results showed that vestitol (IC20 =214.7μM) reduced the expression of genes enrolled with the alpha tubulin (fold -3.7), tubulin in microtubules (fold -3.7), and histone h3 (fold=-3.03), and that treatment with neovestitol (IC20 =102.91μM) downregulated prostaglandin E synthase gene (fold=-3.12), which are considered ideal targets for anticancer therapy. These data open avenues for the study of vestitol and neovestitol as potential promising candidates for anticancer therapy. Toxicological, non-clinical, and clinical validation of the findings presented herein is needed.

Chemical characterization and pharmacological action of Brazilian red propolis

Among the natural products, Brazilian propolis stands out because it is considered a source for the discovery of new bioactive compounds. In the last decade, a new Brazilian propolis has been described, known as red propolis due to its reddish pigment. Since then, numerous studies revealing its major components and their pharmacological activity have been described. Thes, this research had the objective of performing an integrative review of the literature on the biological activity of Brazilian red propolis. Scientific studies in the PubMed, SciElo and LILACS database were reviewed and analyzed using an inductive approach and indirect documentation technique. A total 418 publications were found, and onclusion and exclusion criteria were subsequently applied, selecting a total of 18 articles. The Brazilian propolis presents numerous pharmacological properties, especially the antibacterial, antfugal, antoxidant and anti-inflammatory effects. In addition, the chemical composition of this substance is different from other types of propolis already reported in the literature, presenting flavonoids never before reported. In this way, its chemical characteristics suggest probable therapeutic use for the human organism.

Polyprenylated benzophenone-enriched extracts obtained using SC–CO2 from the dry ethanolic extract of Brazilian red propolis

ABSTRACTRed propolis is a well-known potent antimicrobial source because of its various bioactive compounds. This work aims to submit a sample of Brazilian red propolis to supercritical CO2 extraction, with subsequent chemical characterisation by HPLC and UHPLC–MS, and to evaluate the antifungal activity against three strains of Candida glabrata. The method proved to be selective for the extraction of benzophenones. The results demonstrated a correlation between the presence of benzophenones and antifungal activity. The supercritical extract that seems to be the richest in benzophenones was the one obtained with a pressure of 300 bar and was the most active against the C. glabrata strains.Abbreviations: SFE, Supercritical Fluid Extraction; MIC, Minimal Inhibitory Concentration; SC, Supercritical; BZP, benzophenones

Brazilian Red Propolis: Legitimate Name of the Plant Resin Source

Red propolis ranks as the second most produced Brazilian propolis type Its main chemical constituents are phenolic substances typical of the subfamily Faboideae of the Fabaceae Leguminosae family such as neoflavanoids chalcones flavanones and isoflavonoids among them isoflavones e g biochanin a daidzein and pterocarpans e g medicarpin The plant source of Brazilian red propolis has been referred to as Dalbergia ecastophyllum throughout the propolis literature Such scientific binomial however is a synonym of the legitimate name Dalbergia ecastaphyllum

Chemical Study and Biological Evaluation of the Red Propolis from Alagoas

A investigacao quimica do extrato etanolico de propolis vermelha permitiu o isolamento da 2’-hidroxi-4’,7-dimetoxiisoflavana ( 1 ), da 2’,7-dihidroxi-4’-metoxiisoflavana ( 2 ), da 2’,4’-dihidroxi-7-metoxiisoflavana ( 3 ), da 4’,7-dihidroxi-2’-metoxiisoflavana ( 4 ), da 2’,4’,4-trihidroxichalcona ( 5 ) e do lup-20(29)-em-3-ol ( 6 ). As estruturas das substâncias foram caracterizadas com base nos seus dados espectrais e por comparacao com a literatura. Foi realizado o teste de atividade antioxidante do extrato da propolis, e suas fracoes hexânica, diclorometano, acetato de etila e metanolica, obtendo-se resultados satisfatorios e superiores ao padrao Vitamina C. Outra atividade testada foi a anticolinesterasica, tanto do extrato quanto das fracoes hexânica, diclorometano, acetato de etila e metanolica, obtendo-se resultados positivos em todas as amostras em relacao ao padrao sal de Eserina, exceto para FHEEPV.

Zinc phosphate with red propolis antimicrobial effect and tensile strength

Zinc phosphate with red propolis antimicrobial effect and tensile strength

Antibacterial action of red and green propolis extract in infected root canal

OBJECTIVE: This study investigated the antibacterial action of 30% red propolis, 40% green propolis and 2.5% sodium hypochlorite with irrigation protocols in infected root canals.METHODS: During 60 days, twenty-four root canals were inoculated with E. faecalis. In all experimental groups were performed root canal preparation and two irrigation protocols - passive ultrasonic irrigation and conventional irrigation. In the groups 1, 3, 5 and 7 it were made root canal preparation associated to conventional irrigation with 30% propolis, 40% propolis, 2.5% sodium hypochlorite and distilled water, respectively. In groups 2, 4, 6 and 8 it was made root canal preparation associated with passive ultrasonic irrigation with the solutions described above. The groups 9 and 10 were the controls (negative and positive). Samples of the root canals were collected and immersed in 7 mL of BHI, for 48 hours, at 37ºC. Bacterial growth was analyzed by turbidity of the culture medium.RESULTS: Antibacterial action was observed of 30% red propolis and 2.5% sodium hypochlorite when used passive ultrasonic irrigation and conventional irrigation after 20 minutes, but in 72 hours it was not effective.CONCLUSION: The irrigating agents and protocols were not effective to eliminate the microorganisms of the infected root canals. Clinical Significance: Propolis presents potential for clinical application due to its antimicrobial, anti inflammatory, antioxidant and low toxicity properties.

Evaluation of bioactive compounds potential and antioxidant activity of brown, green and red propolis from Brazilian northeast region

The aim of the present study was to determine the contents of bioactive compounds present in brown, green and red species of propolis cultivated in the Brazilian northeast states of Alagoas and Sergipe. The contents of phenolic compounds, flavonoids and antioxidant activity (DPPH, ABTS+, FRAP, ORAC) were determined. Identification and quantification of phenolic and flavonoid compounds were performed by using UHPLC-QqQ-MS/MS system. The results revealed high contents of total phenolics and flavonoids. Among the three species, the antioxidant potential had higher capacity in the red propolis. The presence of some of bioactive compounds viz. acacetin, artepellin C, eriodictyol, gallic acid, isorhamnetin, protocatechuic acid, vanillin and vanillic acid in Brazilian red propolis is reported for the first time in this work. Positive correlation between total phenolics versus the FRAP and ORAC methods was established which led to conclusion that antioxidant activity of propolis is mainly due to its phenolic compounds.

Mechanisms underlying the cytotoxic effect of propolis on human laryngeal epidermoid carcinoma cells

Propolis has been used as a traditional remedy for centuries because of its beneficial effects, including anticancer properties. The aim of this study was to compare the cytotoxic mechanism of Cuban red propolis (CP) and Brazilian green propolis (BP) on human laryngeal carcinoma (HEp-2) cells. Cell viability, leakage of lactate dehydrogenase, fluorescence staining, mitochondrial membrane potential (ΔΨm) and the expression of pro/anti-apoptotic genes were assessed. Cell viability and cytotoxic assays suggested a dose-dependent effect of CP and BP extracts with a possible association of intracellular reactive oxygen species production and decreased ΔΨm. Both samples induced apoptosis via activation of TP53, CASP3, BAX, P21 signalling, and downregulation of BCL2 and BCL-XL. CP exerted a higher cytotoxic effect than BP extract. Our findings suggest further investigation of the main components of each propolis sample, what may lead to the development of strategies for the treatment of laryngeal cancer.

Anti-inflammatory mechanisms of neovestitol from Brazilian red propolis in LPS-activated macrophages

Neovestitol is considered one of the main bioactive components of Brazilian red propolis. Neovestitol’s antimicrobial and antioxidant effects have already been demonstrated. Here, neovestitol immune modulatory effects were investigated on LPS activated macrophages. RAW264.7 murine macrophages activated with LPS were treated with neovestitol and NO production, cell viability and cytokines profile were determined. Activation of inflammatory signaling pathways and macrophage polarization were determined by RT-qPCR and Western blot. Neovestitol at 0.22µM inhibited NO production by 60% without affecting cell viability and reduced GM-CSF, IFN-γ, IL-1β, IL-4, TNF-α and IL-6 levels, whereas increased IL-10 production. These cytokines profile changes were associated with the downregulation of transcription of genes involved in nitric oxide production, NF-κB, IL-1β, and TNF-α signaling pathways. NF-κB and MAPK signaling pathways inhibition and decreased levels of TIRAP were further confirmed by Western blot. Neovestitol, as a nutraceutical, is a potential candidate to modulate chronic inflammation in humans.