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Artificially induced bone tissue losses after the application of ethanol extract of propolis (EEP) showed an accelerated rate of ossification. The osteogenetic process was just about half as long as in the control group.
The use of ethanol extract of propolis (EEP) on injured dental pulp results in the stimulation of regenerative processes. A reduction of disorders of the circulatory system is observed, inflammatory and degenerative processes are also reduced.
A study consisting of an examination of the acute toxicity of ethanol extract of propolis (EEP) in mice, its effect on spontaneous movement in mice and rats, its analgesic properties and its influence on body temperature in mice was conducted. Also examined was the activity of EEP on animals under the influence of narcotics and spontaneous movement under the influence of amphetamine, its effects on blood pressure and respiration in rats. The results of these examinations indicate that EEP injected i.p. has a weak general effect on the experimental animals.
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AIM: The aim of this study was to determine the antioxidative activity of propolis from ecologically clean parts of Dalmatia. METHODS: Phenol concentration in ethanolic propolis extracts was determined by Folin-Ciocalteu reagent using gallic acid as the standard. Flavonoid phenolic compounds were analyzed after precipitation with formaldehyde. The residual non-flavonoid phenolics were also determined by Folin-Ciocalteu method. By determining the change of peroxide number (PN), of tiobarbiture acid reactive species (TBARS), and of DPPH-radical activity, antioxidative efficiency of propolis was tested and compared with well known and widely used synthetic antioxidants. Values of PN and TBARS were determined at 60 degrees C in samples of trigyceride substrate (lard) without and with the addition of antioxidants. Compared was the efficiency of three antioxidants: propolis (alcoholic extract), vitamin E, and (+)-catechin in a concentration of 1%. PN was monitored during 50 days. By the method of Sedlacek, TBARS were measured during 30 days. Antioxidative activity of propolis extract was also measured in terms of hydrogen donating ability using stable radical alpha,alpha-diphenyl-beta-picril hidrazyl (DPPH*) and compared with commercial synthetic antioxidants of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and (+)-cathecin. Inhibition degree of DPPH* was calculated by the formula of Yen and Duh. RESULTS: Total phenol content, expressed as gallic acid, in propolis extracts varied from 75.2 to 90.2 g/kg propolis. The proportion of flavonoids in total phenols ranged from 62% to 65%. Values of TBARS were not increased only in samples with added propolis. The inhibition of DPPH-radical by propolis extracts ranged from 93% to 96%, by catechin 95%, by BHT 49%, and by BHA 64%. Compared to BHT and BHA, propolis extracts showed greater reducing activity against DPPH-radical. DISCUSSION: The chemical composition of propolis, and thus its biological activity depend on the plant from which it has been collected, and on the macro- and microclimatic conditions. Many compounds in propolis exert antioxidative activity. A belief was expressed that the biological activity of propolis is very probably based mostly on its antioxidative efficiency. Dalmatian propolis showed high efficiency in the prevention of oxidative processes. This could be explained by the high proportion of polyphenol constituents, especially flavonoids. A very low and equal degree of increase of PN, as a measure of oxidative processes, was noticed in the samples of triglyceride substrate with the addition of propolis and (+)-catechin. The greatest rise of TBARS was measured in the samples of pure lard. There was no increase of TBARS only in the samples with added propolis. Propolis and (+)-catechin showed great efficiency in the inhibition of DPPH-radical, greater than BHT and BHA, which are widely used in food industry. CONCLUSION: The results indicate that Dalmatian propolis could be an efficient protective agent against oxidative processes in food. The high antioxidative activity of propolis, its natural origin, and present knowledge about its biological properties, make it a very promising nutritional additive for human diet.
In this study, we focused on immune stimulation by Propolis, and examined changes in the effect of irradiation after Propolis administration. We also examined the radioprotective effect of Propolis by observing its effect on the immune system. The effect of immune activation by Propolis was investigated by measuring the total immunoglobulin (Ig) G and IgM. The radioprotective effect of immune activation by Propolis was investigated by measuring the T-lymphocyte subsets in the peripheral blood of mice following whole body irradiation. Compared with the control group, the IgG was significantly reduced in the Propolis group, indicating that Propolis suppressed IgG production. ELISA revealed that the amount of IgM in mouse serum was significantly higher in the Propolis group as compared with the control group, indicating that Propolis increased IgM production. The number of CD4-positive cells was increased only in the Propolis group. Likewise, the number of CD4-positive cells increased by 81% in the Propolis with irradiation group compared with the irradiation group alone. Compared with the control group, the Propolis group increased CD8-positive cells. Compared with the irradiation alone group, CD8-positive cells were decreased by Propolis with irradiation group. Propolis activated macrophages to stimulate interferon (IFN)-gamma production in association with the secondary activation of T-lymphocytes, resulting in a decrease in IgG and IgM production. Cytokines released from macrophages in mouse peripheral blood after Propolis administration activated helper T-cells to proliferate. In addition, activated macrophages in association with the secondary T-lymphocyte activation increased IFN-gamma production and stimulated proliferation of cytotoxic T-cells and suppressor T-cells, indicating the activation of cell-mediated immune responses.
Propolis exhibits antimicrobial, anti-inflammatory and other biological effects. The aim of this study was to evaluate the activity of 30 % ethanolic extract of Bulgarian propolis against 94 Helicobacter pylori strains by three methods. By the agar-well diffusion method, only 13.8 % of the strains exhibited no inhibition by 30 microl propolis extract (containing 9 mg propolis) and all isolates were inhibited to some extent by 90 microl of the extract (27 mg propolis) per well. The mean diameters of growth inhibition by 30, 60 or 90 microl propolis extract or 30 microl 96 % ethanol per well were 16.8, 19.2, 27.5 and 8.3 mm, respectively. The propolis extract was more active than the ethanol (P < 0.001). With 90 microl propolis extract per well, 69.4 % of the strains exhibited large diameters of growth inhibition (> or =20 mm) versus 26.6 % with 30 mul per well (P < 0.001). With moist propolis discs, inhibition was detected in more strains (92.1 %) than with dried discs (78.2 %, P < 0.05), with mean inhibitory diameters of 18.7 and 13.8 mm, respectively. By the agar dilution method, 100 and 300 microg propolis ml(-1) inhibited the growth of 57.1 % and 76.2 %, respectively, of the 21 strains tested. In conclusion, Bulgarian propolis had a strong and dose-dependent activity against most of the H. pylori strains tested. Although the effect of propolis on H. pylori in vitro is promising, further microbiological, pharmacological and clinical trials are required.
PURPOSE: The aim of this study was to investigate the antimicrobial and anti-inflammatory properties of an ethanolic extract of propolis, a natural resin produced by honeybees, and to determine synergistic activity between ciprofloxacin and propolis in the treatment of experimental Staphylococcus aureus keratitis. METHODS: Sixteen young New Zealand white rabbits were given intrastromal injections of S. aureus strains. Twenty-four hours later, the rabbits were randomly divided into 4 groups: group 1 was treated with topical 0.3% ciprofloxacin drops along with the ethanolic extract of propolis drops; group 2 received topical 0.3% ciprofloxacin drops; group 3 was administered the ethanolic extract of propolis drops, and group 4, the control group, was treated with phosphate-buffered saline (PBS) drops. Drugs were instilled 8 times/day for 72 h. Twenty-four and 96 h after inoculation of bacteria, the eyes were examined by slit lamp to assess corneal opacity. Corneas were removed to count bacteria. RESULTS: Slit lamp examination showed that the corneal opacity scores were significantly lower in eyes that received propolis plus ciprofloxacin than in those treated with ciprofloxacin (p = 0.041) or propolis (p = 0.006) or control eyes treated with PBS (p = 0.0001). There was no significant difference in eyes treated with ciprofloxacin and propolis (p = 1.00). There were significantly fewer bacteria in eyes that received propolis plus ciprofloxacin than in those treated with ciprofloxacin (p = 0.0001) or propolis (p = 0.0001) or control eyes treated with PBS (p = 0.0001). There was no significant difference in eyes treated with ciprofloxacin and propolis (p = 0.38). CONCLUSIONS: Taking these findings into consideration, we suggest that the ethanolic extract of propolis has antimicrobial and anti-inflammatory properties for S. aureus keratitis. The combination of ciprofloxacin and propolis had better therapeutic effects than either agent alone.
Propolis, a honeybee product, has gained popularity as a food and alternative medicine. Its constituents have been shown to exert pharmacological effects, such as anti-microbial, anti-inflammatory and anticancer. Shoot apices of Baccharis dracunculifolia (alecrim plant, Asteraceae) have been pointed out as sources of resin for green propolis. The present work aimed (i) to observe the collecting behavior of bees, (ii) to test the efficacy of histological analysis in studies of propolis botanical origin and (iii) to compare the chemistries of alecrim apices, resin masses and green propolis. Bee behavior was observed, and resin and propolis were microscopically analyzed by inclusion in methacrylate. Ethanol extracts of shoot apices, resin and propolis were analyzed by gas chromatography/mass spectroscopy. Bees cut small fragments from alecrim apices, manipulate and place the resulting mass in the corbiculae. Fragments were detected in propolis and identified as alecrim vestiges by detection of alecrim structures. Prenylated and non-prenylated phenylpropanoids, terpenoids and compounds from other classes were identified. Compounds so far unreported for propolis were identified, including anthracene derivatives. Some compounds were found in propolis and resin mass, but not in shoot apices. Differences were detected between male and female apices and, among apices, resin and propolis. Alecrim apices are resin sources for green propolis. Chemical composition of alecrim apices seems to vary independently of season and phenology. Probably, green propolis composition is more complex and unpredictable than previously assumed.
In this paper, the qualitative and quantitative separation and determination of the polyphenolic component of propolis preparations in the form of ethanolic extract, usually used for commercial pharmaceutical preparations, has been investigated by means of on-line HPLC-ESI/MS technique. Propolis of different origin have been evaluated for their components and a specific fingerprint has been determined potentially useful for the quality control of extracts in pharmaceutical preparations. The ethanolic extracts of propolis from Argentina, Italy and Spain shows approximately the same total ion chromatogram (TIC) profile due to the presence of the same molecular species, identified by the negative ESI-MS. On the contrary, the samples from Azerbaijan, China, Ethiopia and Kenya show a very peculiar TIC profiles. By using many purified flavonoids and calibration curves over a wide concentration range, from 0.05 (5 microg/ml) to 5 microg (500 microg/ml), an accurate assessment of the contents of several bioactive compounds in extract samples was performed. The propolis from Argentina, Italy and Spain show a great amount of pinocembrin (approximately 49%, 48% and 39% of the total identified flavonoids, respectively) and variable but similar percentages of the other species. On the contrary, the propolis from China, Azerbaijan and Ethiopia have a great amount of pinocembrin (approximately 63%, 46% and 62%, respectively) but no presence of genistein, kaempferol, apigenin and chrysin for the sample from China, genistein, kaempferol, acacetin and chrysin for the propolis from Azerbaijan, and no kaempferol and acacetin for the sample from Ethiopia. The ethanolic extract from propolis of Kenya has no identified flavonoid species but just a peak possessing a m/z of 253.0. Finally, an evaluation of the presence of total flavonoids for the various propolis samples was performed, with extracts from Argentina, Italy and Spain more rich in polyphenols than those from Azerbaijan, China, Ethiopia and Kenya. The HPLC-ESI/MS under the experimental conditions illustrated represents a valuable method for the qualitative and quantitative assay of the most relevant components of propolis. On-line HPLC-ESI/MS analysis constitutes an alternative to obtain typical fingerprints of propolis and a reliable identification of a large number of propolis polyphenolic components.
Propolis, or bee glue, which contains a complex mixture of secondary metabolites, has long been used in many countries for the management of several diseases. The purpose of this study was to evaluate, by means of several pharmacological models, the anti-hyperalgesic effect of propolis collected in the south of Brazil. The abdominal constrictions induced in mice by intraperitoneal injection of acetic acid (0.6%), kaolin (50 mg kg-1) or zymosan (40 mg kg-1) were inhibited to different extents by an extract of propolis (1-60 mg kg-1) administered intraperitoneally 30 min earlier; mean ID50 (concentrations resulting in 50% inhibition) values were 2.7, 10.8 and 10.7 mg kg-1, respectively, and maximum inhibition was 58 +/- 5, 57 +/- 10 and 51 +/- 5%, respectively. Given orally (25-200 mg kg-1, 1h previously) propolis also inhibited the abdominal constrictions induced by acetic acid (maximum inhibition 43 +/- 5%). When injected intraperitoneally (3-60 mg kg-1, 30 min previously), propolis attenuated both the neurogenic (first phase) and inflammatory (second phase) pain responses and paw oedema caused by intraplantar injection of formalin (2.5%); maximum inhibition was 32 +/- 5, 43 +/- 6 and 19 +/- 2%, respectively. Oral administration of propolis (25-200 mg kg-1, 1 h previously) inhibited both phases and reduced the oedema formation associated with the second phase of the formalin test (maximum inhibition 22 +/- 5, 33 +/- 6 and 26 +/- 3%) and extract of propolis (3-30 mg kg-1 i.p. or 25-100 mg kg-1 p.o., respectively 30 min and 1 h previously) significantly inhibited capsaicin-induced pain with maximum inhibition of 39 +/- 8 and 41 +/- 8%, respectively. When assessed in the Randall-Sellito test of pain, the extract of propolis (3-30 mg kg-1, i.p., 30 min previously) significantly reversed the hyperalgesia induced by intraplantar injection of bradykinin (3 nmol per paw) in rats (P < 0.01). In contrast with morphine the extract of propolis (< or = 100 mg kg-1, 30 min previously) was ineffective when assessed in the tail-flick and hot-plate thermal assays. Naloxone (5 mg kg-1 i.p.) reversed (P < 0.01) the effect of morphine (5 mg kg-1 s.c.) by 70 and 94% respectively in the first and second phases of the formalin test, but did not interfere with the analgesic effect of propolis (10 mg kg-1 i.p., 30 min previously). These results show that ethanolic extract of propolis, given systemically, has significant anti-hyperalgesic action when assessed in chemical, but not thermal, models of nociception in mice and rats. Its analgesic action seems to be unrelated to release or activation of the opioid system.
This study examines the mechanisms by which the standardised ethanolic extract of propolis induces relaxation of the guinea-pig trachea in-vitro. In guinea-pig trachea with or without epithelium and contracted by histamine, the propolis extract caused reproducible and graded relaxation, with a mean EC50 value of 3.8 or 10.5 microg mL(-1) and Emax of 100%, respectively. The propolis extract-induced relaxation was markedly reduced (26+/-9 and 96+/-3%) when guinea-pig tracheas were exposed to Krebs solution containing elevated K+ in the medium (40 or 80 mM). Pre-incubation of guinea-pig tracheas with tetraethylamonium (100 mM) or with 4-aminopyridine (10mM) reduced the propolis extract-induced relaxation by 31+/-10% and 28+/-2%. Likewise, apamin (0.1 microM), charybdotoxin (0.1 microM) or iberiotoxin (0.1 microM) caused marked inhibition of propolis extract-mediated relaxation in guinea-pig trachea (percentage of inhibition: 65+/-3%, 60+/-5% and 65+/-9%, respectively). Also, glibenclamide (1 microM) inhibited the relaxant response caused by the propolis extract by 57+/-4%. Omega-conotoxin GIVA (0.1 microM) or capsaicin (1 microM) produced small but significant inhibition (30+/-5% or 47+/-7%, respectively) of the propolis extract-induced relaxation. The vasoactive intestinal peptide (VIP) antagonist D-p-Cl-Phe6,Leu17[VIP] porcine (0.1 microM) inhibited relaxation by 55+/-5%, while propranolol (1 microM) induced a parallel rightward displacement (about 20 fold) of the propolis extract concentration-response curve. Finally, the propolis extract-induced relaxation was inhibited by the nitric oxide synthase inhibitor L-N(G)-nitroarginine (L-NOArg, 100 microM) (48+/-6%), and by the soluble guanylatecyclase inhibitormethylene blue (10 microM) (37+/-6%), whilethe moreselectivesoluble guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolol[4,3-alquinoxalin-1-one (ODQ, 1 microM) produced only a parallel (about 3 fold) rightward displacement of the propolis extract concentration-response curve. Collectively, these results support the notion that the propolis extract-mediated relaxation in the guinea-pig trachea involves the release of nitric oxide, probably from sensory neurons, besides the activation of soluble guanylate cyclase and activation of Ca2+- and ATP-sensitive K+ channels. Furthermore, the stimulation of beta2-adrenergic and VIP receptors also seems to account for its relaxant action.
The composition, bacteriostatic and ROO*-scavenging potential activities of fifteen propolis samples from various botanic and geographic origins were determined to obtain objective information related to propolis quality. Variance analysis showed significant differences (p < or = 0.05) in the contents of polyphenols, flavonoids and active components between fresh and aged propolis. The state of the product (fresh or aged) could be differentiate by using flavonoid pattern and biological activities. A minimum propolis concentration of 80 microg/ml was required inhibit Bacillus subtilis and Staphylococcus aureus while 800 microg/ml was required to inhibit Escherichia coli using fresh propolis. Aged propolis inhibit B. subtilis and S. aureus at concentration of 100 microg/ml and E. coli at 1000 microg/ml. A minimum flavonoids percentage of 18 g/100 g and a maximum ROO*-scavenging potential activity of 4.3 microg/ml were determined in fresh propolis. Flavonoids levels in aged propolis were approximately 20% lower than in fresh propolis. A maximum flavonoids percentage of 19.8 g/100 g and a ROO*-scavenging potential activity between 5.7 to 6.4 microg/ml in aged propolis were quantified. Another objective was to assess the use of ROO*-scavenging potential activity in propolis quality.
Propolis is a resinous hive product collected by honeybees from plants. The propolis produced in Southeastern of Brazil is known as green propolis because of its color. Modern herbalists recommend its use because it displays antibacterial, antifungal, antiviral, hepatoprotective, anti-inflammatory, immunomodulatory and anti-ulcer properties. The anti-ulcer activity of green propolis hydroalcoholic crude extract was evaluated by using models of acute gastric lesions induced by ethanol, indomethacin and stress in rats. Moreover, the effects of extract on gastric content volume, pH and total acidity, using pylorus ligated model were evaluated. Animals pretreated with propolis hydroalcoholic crude extract (50, 250 and 500 mg/kg) showed a significant reduction in lesion index, total affected area and percentage of lesion in comparison with control group (p<0.05) in the ethanol-induced ulcer model. Green propolis extract, at a higher dose (500 mg/kg), displayed a significant protection by reducing (p<0.05) the evaluated parameters in the gastric ulceration induced by indomethacin. In the stress-induced ulcer model it was observed a significant reduction (p<0.05) in those parameters in animals treated with green propolis extract (250 and 500 mg/kg). Regarding the pylorus ligated model it was observed that green propolis extract (250 and 500 mg/kg) displayed an anti-secretory activity, which lead to a reduction in the gastric juice volume, total acidity and pH. These findings indicate that Brazilian green propolis displays good anti-ulcer activity, corroborating the folk use of propolis preparations, and contributing for its pharmacological validation.
AIMS: The antibacterial activity against Staphylococcus aureus of honey and propolis produced by Apis mellifera and Tetragonisca angustula was evaluated. Secondary aims included the study of the chemical composition of propolis and honey samples and its relationship with antibacterial activity against S. aureus. METHODS AND RESULTS: The antibacterial activity of honey and propolis was determined by the method of macrodilution. The minimum inhibitory concentration (MICs) of A. mellifera honey ranged from 126.23 to 185.70 mg ml(-1) and of T. angustula from 142.87 to 214.33 mg ml(-1). For propolis, the MIC ranged from 0.36 to 3.65 mg ml(-1) (A. mellifera) and from 0.44 to 2.01 mg ml(-1) (T. angustula). Honey and propolis were evaluated by high-performance liquid chromatography. Some typical compounds of Brazilian propolis were also identified in honey samples. Principal component analysis revealed that the chemical composition of honey and propolis samples was distinct based on the geographical location of the samples. CONCLUSIONS: Propolis samples had higher antibacterial activity against S. aureus when compared with honey. However, both propolis and honey samples had antibacterial against S. aureus. SIGNIFICANCE AND IMPACT OF THE STUDY: These antimicrobial properties would warrant further studies on the clinical applications of propolis and honey against S. aureus.