Ethanol Extract Of Green Propolis Research Articles

Oral Films Printed with Green Propolis Ethanolic Extract.

Oral film (OF) research has intensified due to the effortless administration and advantages related to absorption in systemic circulation. Chitosan is one of the polymers widely used in the production of OFs; however, studies evaluating the maintenance of the active principles' activity are incipient. Propolis has been widely used as an active compound due to its different actions. Printing techniques to incorporate propolis in OFs prove to be efficient. The objective of the present study is to develop and characterize oral films based on chitosan and propolis using printing techniques and to evaluate the main activities of the extract incorporated into the polymeric matrix. The OFs were characterized in relation to the structure using scanning and atomic force electron microscopy; the mechanical properties, disintegration time, wettability, and stability of antioxidant activity were evaluated. The ethanolic extract of green propolis (GPEE) concentration influenced the properties of the OFs. The stability (phenolic compounds and antioxidant activity) was reduced in the first 20 days, and after this period, it remained constant.

Synergistic Effect of Baccharis dracunculifolia DC and Green Propolis with Antimicrobial Drugs against Staphylococcus aureus

Abstract: Relevance: This study demonstrated the synergism of various natural preparations in combination with antibiotics, which is a matter of high potential therapeutic interest due to the de- velopment of bacterial resistance to these antibiotic drugs. This synergism is very relevant because it demonstrates that these extracts and their association with these antibiotics may be viable thera- peutic options against the pathogens that are responsible for a high incidence of nosocomial and community infections. Aims: The study investigated the antibacterial activity against Staphylococcus aureus isolates (methicilin-sensitive Staphylococcus aureus (MSSA) and methicilin-resistant Staphylo- coccus aureus (MRSA)) of green propolis and Baccharis dracunculifolia DC. ethanolic extracts com- bined with classical antibiotics. Methods and Results: In the test of the antibacterial activity of the combination of classical antibiotics with natural extracts (ten antibiotics plus one association of two antimicrobial disc diffusions), 25 isolates were used. The isolates were inoculated in Müeller–Hinton agar culture medium containing the green propolis and Baccharis dracunculifolia DC. extracts. Bac- charis dracunculifolia DC ethanolic extract (BDEE) showed higher antibacterial synergistic behavior with the antibiotics tested than green propolis ethanolic extract (PVEE). Conclusions: BDEE was the preparation that showed the best synergistic antimicrobial effect in sub-doses with the tested anti- biotics. These activities may be related to Artepillin C, the major compound in PVEE, and nerolidol, the major compound in BDEE, in synergism with other minority compounds and related antibiotics.

Hydroxypropyl methylcellulose orally disintegration films produced by tape casting with the incorporation of green propolis ethanolic extract using the printing technique

Green propolis ethanol extract (GPEE) is one of the most used compounds in folk medicine. Alternative dosage forms, such as orally disintegrating films (ODFs) and forms of incorporation of active principles, such as the printing technique, are arousing interest due to the possibility of producing personalized dosages and possible reduction of the degradation of compounds. The aim of this study was to evaluate the effect of green propolis ethanol extract (GPEE) incorporation on hydroxypropyl methylcellulose (HPMC) using the printing technique (1–4 print layers). GPEE was characterized by apparent viscosity, density, and surface tension. The ODFs with different numbers of printed layers were characterized in relation to color, moisture content, mechanical properties, structure, disintegration time, surface pH, stability, cytotoxicity, and anti-inflammatory activity tests. GPEE showed properties similar to those observed for commercial inks. The number of printed layers affected film properties. ODFs with GPEE showed anti-inflammatory activity, where a reduction of about 40% NO 2 − concentration against macrophage cells was found after adding one GPEE layer. As such, printing on hydroxypropyl methylcellulose oral films as a vehicle for green propolis ethanolic extract represents an interesting alternative to produce materials with controlled anti-inflammatory activity. • Viscosity, density and surface tension values of GPEE are similar to commercial inks. • Oral films of HPMC-based printed are a promising way to deliver active compounds. • HPMS-based printed films maintain the stability of phenolic compounds. • Films printed with green propolis ethanol extract have anti-inflammatory activity.

Taiwanese green propolis ethanol extract promotes adipocyte differentiation and alleviates TNF-α-mediated downregulation of adiponectin expression

Abstract The objective was to examine the effects of Taiwanese green propolis (TGP) ethanol extract on adipocyte differentiation and lipolysis. TGP ethanol extract significantly enhanced the differentiation of murine mesenchymal stem cells into adipocytes, and this was accompanied by an increase in intracellular triglyceride content and adiponectin levels. TGP ethanol extract significantly increased the expression of adipogenic transcription factor-associated genes in murine mesenchymal stem cells after hormonal induction of adipogenesis. TGP ethanol extract was able to alleviate norepinephrine-induced lipolysis in differentiated adipocytes, which was accompanied by a decrease in lipolytic gene expression. Besides, TGP ethanol extract could reduce the tumor necrosis factor-α (TNF-α)-mediated inhibition of adiponectin gene expression and secretion during adipogenic differentiation of mesenchymal stem cells and in differentiated adipocytes. These results demonstrate that TGP ethanol extract enhances adipocyte differentiation and is capable of reversing the inhibitory effects of TNF-α on adipocyte differentiation and adiponectin expression in differentiated adipocytes.

Susceptibility of Malassezia pachydermatis Clinical Isolates to Allopathic Antifungals and Brazilian Red, Green, and Brown Propolis Extracts.

Clinical mycoses treatment is associated with issues such as negative side effects, high cost, prolonged treatment, and resistant strain selection. Malassezia pachydermatis is the most frequently isolated yeast in cases of canine otitis and dermatitis. The number of fungal strains exhibiting primary resistance to several drugs in vitro is increasing. Propolis has a diverse chemical composition and well-known therapeutic properties against mycoses. An alternative method for producing propolis extracts using supercritical fluid has higher selectivity, yielding extracts with fewer pollutant residues. This study therefore aimed to evaluate the in vitro susceptibility profile of M. pachydermatis clinical isolates to precharacterized supercritical and ethanolic extracts. Three types of Brazilian propolis extracts (green, red, and brown) and commercial allopathic antifungals were used in this investigation. We used the microdilution broth technique to evaluate the susceptibility profile of the yeasts. The minimum inhibitory concentration (MIC) of the brown propolis ethanolic extract was ≥16 μg/mL for all isolates. The MICs of fluconazole, ketoconazole, itraconazole, and amphotericin B ranged from 8 to >64 μg/mL, 0.032–4 μg/mL, 0.0313–16 μg/mL, and 1–2 μg/mL, respectively. The MICs of ethanolic red propolis extracts were lower than those of supercritical red propolis extracts. However, the green propolis ethanolic extract had more pronounced fungicidal activity. Isolates with lower susceptibility to commercial fungicides were inhibited by red and green propolis extracts. These results indicate that propolis can potentially be used in in vivo experiments as a promising therapeutic agent against M. pachydermatis infections.

Effect of Brazilian green propolis on microorganism contaminants of surface of Gorgonzola-type cheese.

Blue cheeses are susceptible to yeast and bacterial growth on their surface, which causes spoilage during ripening process and the formation of slime. The dairy industry frequently control the proliferation of undesirable microorganisms with natamycin and high salt concentration. The green propolis is a complex of substances that presents antimicrobial properties with great potential as preservative in the food industry. The aims of the present study were to identify the mesophilic aerobic microorganisms present on the surface of Gorgonzola-type cheese, evaluate the antifungal and antibacterial effects of the ethanol extract of green propolis (EEP) on the development of those microorganisms and verify the effects of EEP on the sensory quality of cheese. Ten yeast species belonging to genera Yarrowia, Candida, Debaryomyces and Saccharomyces were identified, as well as seven species of bacteria belonging to genera Staphylococcus, Bacillus, Enterococcus, Corynebacterium and Proteus. The EEP showed minimum biocide concentration (MBC), between 0.3% (weight/weight) and 5% for Bacillus cereus and Proteus vulgaris, respectively. Saccharomyces cerevisiae was the most sensitive species (MBC of 0.63%) and Candida parapsilosis the most resistant one (MBC of 5%). In the sensory analysis, the cheeses involved with EEP at 5% concentration did not differ from the control, while at 10%, there was a slight decrease in acceptance. The EEP has potential and feasibility to be used in Gorgonzola-type cheese, inhibiting the main bacteria and yeasts without affecting largely the sensory characteristics of the product.

Taiwanese Green Propolis Ethanol Extract Delays the Progression of Type 2 Diabetes Mellitus in Rats Treated with Streptozotocin/High-Fat Diet.

Taiwanese green propolis ethanol extract (TGPE) is produced only in Taiwan and has a different composition from other types of propolis. TGPE is known for its anti-inflammation, anti-oxidation, and anti-microbial properties, but the effects and mechanisms of TGPE in the modulation of diabetes are unclear. In this study, we investigated the effects of TGPE on type 2 diabetes mellitus (T2DM) in a streptozotocin/high-fat-diet (STZ/HFD)-induced T2DM rat model. The results revealed that TGPE delayed the development and progression of T2DM and reduced the severity of β-cell failure. TGPE also attenuated inflammation and reactive oxygen species ROS in the rats. Moreover, there were higher levels of oxidant cytokines, leptin, and adiponectin in the serum of the TGPE-treated group. Unlike Brazilian propolis, TGPE promoted hepatic genes PPAR-α and CYP7A1, which were related to lipid catabolism and removal. TGPE may thus delay the progression of T2DM through anti-inflammation effects, anti-oxidation effects, and balancing lipid metabolism. It is suggested that TGPE can be a potential alternative medicine for T2DM.

Artepillin C and phenolic compounds responsible for antimicrobial and antioxidant activity of green propolis and Baccharis dracunculifolia DC.

This study investigates the antimicrobial activity in Staphylococcus aureus isolates (methicillin-sensitive S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA)) and antioxidant activity of green propolis, Baccharis dracunculifolia DC extracts and Artepillin C™. The amount of Artepillin C in different extracts was determined by high performance liquid chromatography analysis. Minimum inhibitory concentration 90 (MIC90) was determined using 40 isolates of S. aureus inoculated in Müeller-Hinton agar culture medium containing the green propolis and B. dracunculifolia DC extracts. PVEE (green propolis ethanolic extract) and BDEH (B. dracunculifolia hexanic extract) showed the greatest antimicrobial activity with MIC90 values of 246·3 and 295·5μgml-1 respectively. Green propolis ethanolic and hexanic extracts (PVEE and PVEH respectively) showed the greatest antioxidant activity assessed by DPPH (1,1-diphenyl-2-picryl hydrazyl radical) with IC50 values of 13·09 and 95·86μgml-1 respectively. Green propolis ethanolic displays better antimicrobial and antioxidant activities compared to other extracts. These activities may be related to the presence of Artepillin C in synergism with the other constituents of the extracts. In this study, the antimicrobial activity of the extracts of green propolis and B. dracunculifolia DC demonstrated in MRSA and MSSA clinical isolates indicated that they can be important tools to treat infections caused by these bacteria.

Brazilian propolis extract increases leptin expression in mouse adipocytes.

We investigated the anti-obesity effects of Brazilian green propolis ethanol extract using a mouse model of obesity. Repeated intraperitoneal injection of propolis (100 mg/kg twice a week) caused feeding suppression in C57BL/6 mice, whereas this treatment had negligible effects on C57BL/6 ob/ob mice. Since C57BL/6 ob/ob mice have a missense mutation in the Lep gene, leptin is likely to contribute to the propolis-induced feeding suppression. We found that propolis treatment indeed clearly increased leptin mRNA production in the visceral adipose tissues. Moreover, propolis extract directly elevated leptin expression in differentiated 3T3-L1 adipocytes. Artepillin C, an important organic compound found in Brazilian green propolis, failed to induce leptin mRNA in 3T3-L1 cells. Compounds other than artepillin C in Brazilian propolis must thus cause leptin induction in adipocytes, possibly resulting in the suppression of feeding and obesity.

Influence of the Toothpaste with Brazilian Ethanol Extract Propolis on the Oral Cavity Health

Propolis-based therapeutic agents represent this potential for the development of new drugs in dental care. The aim of a clinical-cohort study was to determine the influence of application of toothpaste enriched with Brazilian extract of propolis (EEP) on health status of oral cavity. Laboratory analysis was conducted in order to assess the chemical composition of EEP including total phenolic compounds, the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity, ABTS radical cation scavenging activity, and FRAP assay. Clinical research involved two groups of subjects comprising 32 adult patients, with assessment based on the preliminary evaluation of the state of their marginal periodontium. The investigation of oral health indices API, OHI, and SBI and microbiological examination of oral microflora were also carried out. Results obtained indicated time-dependent microbial action of EEP at 50 mg/L concentration, with antimicrobial activity against Gram-positive bacteria. The total decrease of API, OHI, and SBI mean values was observed. Hygienic preparations with 3% content of Brazilian ethanol extract of green propolis (EEP) efficiently support removal of dental plaque and improve the state of marginal periodontium.

Propolis: a natural product as an alternative for disinfection of embryonated eggs for incubation

During the cooling process of embryonated eggs, there is a natural air flux from the surface to the inner part of the eggs, carrying contaminants such as bacteria and fungi through the shell's pores, infecting embryos and resulting in the inability to hatch or poor chick quality. Formaldehyde, a toxic product, is still the most used disinfectant for embryonated eggs in the aviculture industry. In order to evaluate the antimicrobial activity of the green propolis ethanolic extract as an alternative to formaldehyde, 140 hatching eggs from laying hens were collected and submitted to disinfection with five different treatments: T1 - without disinfection; T2 - formaldehyde fumigated eggs; T3, T4 and T5 disinfection by immersion in propolis solution in the concentrations of 2,400 µg, 240 µg and 24 µg, respectively. The contamination levels by total mesophiles and fungi of the egg shells (Aspergillus sp. and other moulds) after disinfection with propolis were lower than when compared to the control without disinfection. In comparison with formaldehyde, the 240 µg and 24 µg propolis concentrations did not differ regarding antibacterial activity, but for antifungal activity the 2,400 µg and 240 µg concentrations were more efficient. The 2,400 µg and 240 µg propolis treatments presented a hatching rate of 94.1%, compared to only 84.6% for the formaldehyde treatment. The green propolis ethanolic extract presented antibacterial and antifungal activities in embryonated eggs showing that it can be a new natural disinfectant product substituting formaldehyde.

Protective Effect of Brazilian Propolis Against Hepatic Oxidative Damage in Rats with Water‐immersion Restraint Stress

In the present study we examined the protective effect of Brazilian propolis against hepatic oxidative damage in rats with water-immersion restraint stress (WIRS) in comparison with that of vitamin E (VE). Fasted rats orally received Brazilian green propolis ethanol extract (BPEE; 10, 50 or 100 mg/kg), VE (250 mg/kg) or vehicle at 30 min before the onset of WIRS. Exposure of vehicle-treated rats to 6 h of WIRS caused liver cell damage, judging from the levels of serum alanine aminotransferase and aspartate aminotransferease, increased hepatic lipid peroxide, NO(x) contents and myeloperoxidase activity, and decreased hepatic non-protein SH, ascorbic acid contents and superoxide dismutase activity. Preadministration of BPEE (50 or 100 mg/kg) or VE to the stressed rats protected against the hepatic damage and attenuated the increased hepatic lipid peroxide and NO(x) contents and myeloperoxidase activity and the decreased hepatic non-protein SH and ascorbic acid contents and superoxide dismutase activity. These protective effects of BPEE (50 mg/kg) were greater than those of BPEE (100 mg/kg) and were almost equal to those of VE. These results indicate that BPEE protects against hepatic oxidative damage in rats exposed to WIRS possibly through its antioxidant and antiinflammatory properties such as VE.

Green propolis extract as additive in the diet for lambs in feedlot

The objective of this paper is to assess the effects of the inclusion of different levels of green propolis extract in the diet of lambs in feedlot on ingestive behavior, nutrients digestibility, physiological parameters and performance. Eight lambs were distributed in double Latin Square with four treatments, corresponding to the inclusion levels (4, 8, 12, 16 mL) of green propolis ethanolic extract (30 g of ground crude propolis was infused in a 100-mL hydroalcoholic solution, 700 mL/L). The diets were composed of Brachiaria brizantha cv. MG5 hay and a commercial concentrate (roughage:concentrate ratio was 50:50) in a dry matter basis. No effect was observed on dry matter (31.2 g/kg of BW), crude protein, ether extract, neutral detergent fiber, non-fibrous carbohydrates and total digestible nutrients content (TDN) intakes. No significant effect was seen on the digestibility coefficients, presenting an average of 65.94% of TDN. The green propolis extract levels do not have a significant effect on behavior or physiologic parameters. Seeking to maximize feeding efficiency, the inclusion of 7.60 mL/day (2.1189 mg of dry matter and 0.1123 mg of flavonoids) of green propolis extract in the diet of lambs in feedlot is recommended.

3,4-Dicaffeoylquinic Acid, a Major Constituent of Brazilian Propolis, Increases TRAIL Expression and Extends the Lifetimes of Mice Infected with the Influenza A Virus.

Brazilian green propolis water extract (PWE) and its chemical components, caffeoylquinic acids, such as 3,4-dicaffeoylquinic acid (3,4-diCQA), act against the influenza A virus (IAV) without influencing the viral components. Here, we evaluated the anti-IAV activities of these compounds in vivo. PWE or PEE (Brazilian green propolis ethanol extract) at a dose of 200 mg/kg was orally administered to Balb/c mice that had been inoculated with IAV strain A/WSN/33. The lifetimes of the PWE-treated mice were significantly extended compared to the untreated mice. Moreover, oral administration of 3,4-diCQA, a constituent of PWE, at a dose of 50 mg/kg had a stronger effect than PWE itself. We found that the amount of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) mRNA in the mice that were administered 3,4-diCQA was significantly increased compared to the control group, while H1N1 hemagglutinin (HA) mRNA was slightly decreased. These data indicate that PWE, PEE or 3,4-diCQA possesses a novel and unique mechanism of anti-influenza viral activity, that is, enhancing viral clearance by increasing TRAIL.

Brazilian Green Propolis: Effects In Vitro and In Vivo on Trypanosoma cruzi

The composition of a Brazilian green propolis ethanolic extract (Et-Bra) and its effect on Trypanosoma cruzi trypomastigotes and other pathogenic microorganisms have already been reported. Here, we further investigated Et-Bra targets in T. cruzi and its effect on experimental infection of mice. The IC50/4 days for inhibition of amastigote proliferation was 8.5 ± 1.8 μg mL−1, with no damage to the host cells. In epimastigotes Et-Bra induced alterations in reservosomes, Golgi complex and mitochondrion. These effects were confirmed by flow cytometry analysis. In trypomastigotes, Et-Bra led to the loss of plasma membrane integrity. The in vitro studies indicate that Et-Bra interferes in the functionality of the plasma membrane in trypomastigotes and of reservosomes and mitochondrion in epimastigotes. Acutely infected mice were treated orally with Et-Bra and the parasitemia, mortality and GPT, GOT, CK and urea levels were monitored. The extract (25–300 mg kg−1 body weight/day for 10 days) reduced the parasitemia, although not at significant levels; increased the survival of the animals and did not induce any hepatic, muscular lesion or renal toxicity. Since Et-Bra was not toxic to the animals, it could be assayed in combination with other drugs. Et-Bra could be a potential metacyclogenesis blocker, considering its effect on reservosomes, which are an important energy source during parasite differentiation.

Inhibitory activity of Brazilian green propolis components and their derivatives on the release of cys-leukotrienes

The effects of Brazilian green propolis ethanol extract on Cry j1-induced cys-leukotrienes and histamine release from peripheral leukocytes of patients with allergic rhinitis were investigated. One of the key mechanisms for the anti-allergic properties of the extract was revealed to be the suppression of cys-LTs release. Furthermore, a series of propolis components and their phenethyl esters were synthesized and evaluated as inhibitors of cys-LTs release. Artepillin C, baccharin, and kaempferide were the major active components of the ethanol extract. The inhibitory activity of artepillin C phenethyl ester was comparable to that of existing LT synthesis inhibitors.

Um marcador químico de fácil detecção para a própolis de Alecrim-do-Campo (Baccharis dracunculifolia)

In the present work a volatile chemical marker (CM) for the Baccharis dracunculifolia (Bd) propolis is proposed, which is easily detectable by gas chromatography. It is the most abundant volatile compound in dichloromethane extracts of green propolis from this plant, but it appears also, in different concentrations, in dichloromethane extracts of brown, dark and red propolis from regions where Bd grows. The CM is present in significative concentration in the bud extract of Bd, in contrast to the leaf extract where its concentration is low. Propolis from regions without Bd does not contain the CM. This compound was recently isolated; it is the allyl 3-prenylcinnamate. Commercial samples of green propolis ethanol extract were analyzed and the first quality one (exportation standard) presented the highest concentration on CM. This finding makes easier the quality control of green propolis extracts sold at the market.

Immunomodulation produced by a green propolis extract on humoral and cellular responses of mice immunized with SuHV-1

Despite recent technological advances in vaccine production, most the vaccines depend on the association with adjuvant substances. This work evaluated the adjuvant capacity of an ethanol extract of green propolis associated to inactivated Suid herpesvirus type 1 (SuHV-1) vaccine preparations. Mice inoculated with SuHV-1 vaccine plus aluminum hydroxide and 5 mg/dose of propolis extract presented higher levels of antibodies when compared to animals that received the same vaccine without propolis. The use of SuHV-1 vaccine with propolis extract alone did not induce significant levels of antibodies, however it was able to increase the cellular immune response, evidenced by the increase in the expression of mRNA to IFN-γ. Besides, propolis increased the percentage of protected animals against challenge with a lethal dose of SuHV-1. The effect of green propolis extract on the humoral and cellular immune responses may be exploited for the development of effective vaccines.

Effect of Brazilian green propolis on the production of reactive oxygen species by stimulated neutrophils

The activity of a crude ethanol extract of green propolis and its fractions obtained by partition with hexane, chloroform and n-butanol was assessed on luminol- and lucigenin- enhanced chemiluminescence (CL) produced by rabbit neutrophils (PMNs) stimulated with particles of serum-opsonized zymosan (OZ). The total production of reactive oxygen species (ROS) by PMNs was measured by the luminol-enhanced CL (LumCL) assay and the production of the superoxide anion (O 2 −) by the lucigenin-enhanced CL (LucCL) assay. All evaluated propolis samples had inhibitory effect on the LumCL and LucCL, which was concentration dependent. The n-butanol and chloroform fractions displayed the highest inhibitory effect on the LumCL produced by PMNs stimulated with OZ, in comparison with both the ethanol extract and the hexane fraction. Besides, the hexane fraction was the one which presented the highest effect for the LucCL assay. Some isolated compounds from both n-butanol and chloroform fractions were also assessed, including kaempferide, isosakuranetin, aromadendrine-4′-methyl-ether and 3-prenyl- p-coumaric acid. Kaempferide presented the highest inhibitory effect on the LumCL in comparison with the other compounds. Moreover, under the conditions assessed, the studied green propolis samples and isolated compounds were not toxic to the rabbit PMNs.