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Pulmonary carcinogenesis induced by ferric nitrilotriacetate in mice and protection from it by Brazilian propolis and artepillin C.

In experiments using the renal carcinogen ferric nitrilotriacetate (Fe-NTA) in male ddY mice, primary pulmonary cancers were also induced in bronchiolar and alveolar tissues. 4-Hydroxy-2-nonenal (4-HNE) and 8-hydroxy-2'-deoxyguanosine (8-OHdG), products of oxidative processes, increased in bronchiolar and alveolar cells after administration of Fe-NTA. These substances disappeared after oral administration of propolis or artepillin C, as shown histochemically, and correlated with an anticancer prophylactic effect of propolis and artepillin C. From our investigation, lipid peroxidation seems to play an important role in pulmonary carcinogenesis. Malignant progression from adenoma of bronchiolar or alveolar origin to malignant tumors has been proposed to involve a stepwise transformation. In our study, adenomas developed into adenocarcinomas and large cell carcinomas after treatment with Fe-NTA. In contrast, after oral administration of propolis or artepillin C, adenomas did not progress to carcinomas. Instead of developing into large cell cancers, as induced by Fe-NTA in control mice, adenomas showed remarkable proliferation of macrophages and local anti-oxidant activity after treatment with either propolis or artepillin C. Propolis and artepillin C therefore appear to inhibit lipid peroxidation and the development of pulmonary cancers.

8-Hydroxy-2'-Deoxyguanosine↗

Scavenging action of propolis extract against oxygen radicals.

The ethanolic extracts of two types of cuban propolis (R and P) showed a similar manner of scavenging action against different species of oxygen radicals which were generated by specific chemical reactions. Chemiluminescence produced by superoxide generated from the xanthine-xanthine oxidase reaction was 50% inhibited by approximately 5 micrograms/ml of propolis R and 9.5 micrograms/ml of propolis P and by catechin (0.15 micrograms/ml) and superoxide dismutase (72 ng/ml). Alkoxy radical scavenging effect was similar to that produced by 0.11 micrograms/ml of alpha-tocopherol: inhibition of chemiluminescence by 50% was caused by approximately 0.6 micrograms/ml of both propolis preparations. The results indicate that the antioxidative properties of both propolis could be attributed to their free radical scavenging activity against alkoxy radicals and to a lesser degree against superoxide.

Catechin↗

Effect of different formulations of propolis on mice infected with Trypanosoma cruzi.

Propolis, a bee product, can be regarded as one of the potential natural sources in folk medicine, displaying strong antimicrobial activity. Previous work showed that propolis extracts exhibited in vitro activity against Trypanosoma cruzi (Higashi and de Castro, 1994). Different formulations of propolis were administered to experimentally Trypanosoma cruzi-infected mice and the parasitemia kinetics and survival rate were monitored. The oral administration of ethanolic extracts up to 1.2 g propolis/kg per day or propolis offered ad libitum in the drinking water (up to 4 g/kg per day) or added to the food (up to 5 g/kg per day) did not interfere with both parameters. The differences between in vitro and in vivo trypanocidal activity of propolis and future perspectives are discussed.

Administration, Oral↗

Effects of Brazilian and Bulgarian propolis on bactericidal activity of macrophages against Salmonella Typhimurium.

Propolis has been used in folk medicine since ancient times due to its many biological properties, such as antimicrobial, antiinflammatory, antioxidant, immunomodulatory activities, among others. Macrophages play an important role in the early phase of Salmonella infection. In this work, macrophages were prestimulated with Brazilian or Bulgarian propolis and subsequently challenged with Salmonella Typhimurium at different macrophage/bacteria ratio. After 60 min of incubation, cells were harvested with Triton-X to lyse the macrophages. To assess the bactericidal activity, the number of colony-forming units (CFU) of S. typhimurium was determined by plating 0.1 mL in Mueller Hinton agar. After 24 h, CFU were counted, and the percentage of bactericidal activity was obtained. Propolis from Brazil and Bulgaria enhanced the bactericidal activity of macrophages, depending on its concentration. Brazilian propolis seemed to be more efficient than that from Bulgaria, because of their different chemical composition. In Bulgaria, bees collect the material mainly from the bud exudate of poplar trees, while in Brazil, Baccharis dracunculifolia DC. was shown to be the main propolis source. Our data also showed that the increased bactericidal activity of macrophages involved the participation of oxygen (H(2)O(2)) and nitrogen (NO) intermediate metabolites.

Animals↗

Effects of Turkish pollen and propolis extracts on respiratory burst for K-562 cell lines.

Bee-collected pollen and propolis are apicultural products which are composed of nutritionally valuable substances and contain considerable amounts of polyphenol substances which may act as potent antioxidants. We wanted to show if respiratory burst within a cancer cell lines could be influenced when incubated with pollen and propolis extracts or not. Pollen and propolis extracts at concentrations of 50, 25, 12.5 and 0 mg/ml were prepared by dimethyl sulfoxide (DMSO). K-562 cell cultures and mononuclear cell (MNC) cultures prepared from a peripheral blood sample to serve as control cells were incubated with extracts for 24 h. Determination of respiratory burst was carried out by intracellular dichlorofluorescein (DCFH) test by using flow-cytometric fluorescence analysis. While about 90% and 66% fluorescence was detected at zero concentrations for both K-562 and MNC cultures, fluorescence positivity decreased (between 3.8% and 11.8%) as concentrations of both propolis and pollen extracts increased for K-562 cell culture, but unchanged (between 20% and 83%) for MNC culture. It was concluded that pollen and propolis extracts inhibit respiratory burst within cancer cell lines probably by their antioxidant potentials.

Cells, Cultured↗

Analysis of propolis from Baccharis dracunculifolia DC. (Compositae) and its effects on mouse fibroblasts.

This paper confirms Baccharis dracunculifolia DC. (Compositae) as the main botanical source of the propolis from southeastern Brazil (state of São Paulo) investigated to ascertain specific biological activity in relation to mouse NIH-3T3 fibroblasts, skin cells directly involved in the cicatrization processes. Flavonoid and total phenolic compounds were determined by spectrophotometry, and chemical composition by HPLC; the chromatographic profile, characterized largely by flavonoids and aromatic acids, was found to be qualitatively similar to that of Baccharis dracunculifolia DC. The adsorption of phenolic compounds in the propolis to skin powder was also investigated, and 68% of these compounds adsorbed to the skin powder. At concentrations from 0.12 to 7.81 microg/ml, the propolis revealed no statistical significant differences from its control solutions; however, at concentrations of 31.25 microg/ml or more, the propolis was toxic to NIH-3T3 cells. Thus, the propolis from Baccharis dracunculifolia DC. (Compositae) presents an in vitro concentration-dependent toxicity on mouse NIH-3T3 fibroblasts.

Animals↗

Therapeutic effect of paclitaxel and propolis on lipid peroxidation and antioxidant system in 7,12 dimethyl benz(a)anthracene-induced breast cancer in female Sprague Dawley rats.

Breast cancer is one of the most common cancers in women of developed and developing countries. The optimum management of which requires a multidisciplinary approach including the use of certain biochemical and molecular markers. The effect of propolis along with paclitaxel on 7,12 dimethyl benz(a)anthracene (DMBA) induced experimental breast cancer was investigated in female Sprague Dawley rats. Female Sprague Dawley rats were divided into five groups of six animals each. Group I served as normal control animal. Group II animals received DMBA (20 mg in 0.5 ml sunflower oil and 0.5 ml of saline) i.p. to develop mammary tumor by the end of 90 days. Group III were breast cancer animals treated with 33 mg paclitaxel/kg body weight (bw) weekly once for 4 weeks. Group IV were breast cancer-bearing animals treated with 50 mg propolis/kg bw for 30 days. Group V were breast cancer-bearing animals treated with both paclitaxel and propolis as mentioned above. Administration of paclitaxel and propolis effectively suppressed breast cancer, which is revealed by the decrease in the extent of lipid peroxidation (LPO) with concomitant increase in the activities of enzymic antioxidants (superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)) and non-enzymic antioxidants (reduced glutathione (GSH), Vitamin C and Vitamin E) levels when compared to breast cancer-bearing animals treated with either paclitaxel or propolis alone. From our results, we conclude that propolis is a potent antioxidant and, when given in combination with paclitaxel, offers maximum protection against DMBA induced mammary carcinogenesis.

9,10-Dimethyl-1,2-benzanthracene↗

Effect of propolis on endotoxin-induced uveitis in rabbits.

PURPOSE: To test the anti-inflammatory effect of propolis, a natural bee-produced compound, and compare it with corticosteroids for the treatment of endotoxin-induced uveitis (EIU). METHODS: EIU was produced in all rabbits by unilateral intravitreal injection of 2,000 ng Salmonella typhimurium endotoxin. The animals were then divided randomly into three groups as follows: group A received no treatment (control); group B received methylprednisolone (5 mg/0.1 mL) (positive control); and group C received propolis (5 mg/0.16 mL) by anterior sub-Tenon injection at the time of uveitis induction and at 4 and 8 hours after induction. Inflammation was evaluated by clinical manifestations and by measuring the protein concentration and inflammatory cell content of the aqueous humor. RESULTS: The clinical grade, cell count, and protein levels in the aqueous humor were: control group (6.0 +/- 0.8, 2,519 +/- 470 cells/microL, 32.9 +/- 2.4 mg/mL); methylprednisolone group (1.8 +/- 0.7, 572 +/- 137 cells/microL, 15.2 +/- 1.8 mg/mL); and propolis group (2.3 +/- 0.5, 503 +/- 124 cells/microL, 13.8 +/- 1.5 mg/mL). Statistically significant differences were recorded in the treatment groups when compared to the control group (P < .001). The effects of methylprednisolone and propolis on EIU were similar (P > .05). CONCLUSIONS: Propolis showed significant anti-inflammatory effects on EIU in rabbits. The mechanism of its action warrants further investigation.

Animals↗

Gelatin microparticles containing propolis obtained by spray-drying technique: preparation and characterization.

Gelatin microparticles containing propolis extractive solution (PES) were prepared by spray-drying technique. The optimization of the spray-drying operating conditions and the proportions of gelatin and mannitol were investigated. Regular particle morphology was obtained when mannitol was used, whereas mannitol absence produced a substantial number of coalesced and agglomerated microparticles. Microparticles had a mean diameter of 2.70 microm without mannitol and 2.50 microm with mannitol. The entrapment efficiency for propolis of the microparticles was upto 41% without mannitol and 39% with mannitol. The microencapsulation by spray-drying technique maintained the activity of propolis against Staphylococcus aureus. These gelatin microparticles containing propolis would be useful for developing intermediary or eventual propolis dosage form without the PES' strong and unpleasant taste, aromatic odour, and presence of ethanol.

Chromatography, High Pressure Liquid↗

Antioxidant activity of Argentine propolis extracts.

Propolis is used in Argentine folk medicine. We have examined its possible protective action against oxidative modification of lipid in unfractionated serum. The kinetics of copper-induced oxidation was continuously monitored by measuring the formation of conjugated dienes, as the increase in the absorbance at 234 nm. According to the kinetics of oxidation, the propolis were classified in three different groups. Group I (CE, CO, BO, MO, BE) inhibited lipid oxidation during the initiation and propagation phases even at low concentrations. Group II (SP, CA, AM) increased the lag-phase for conjugated diene formation. All propolis in groups I and II diminished the maximal rate of diene production and the maximal amount of dienes produced. Group III (PA, RA, FE, VR, TV) had no effect on the lipid oxidation. The extent of lipoprotein oxidation was measured by the thiobarbituric acid reactive substance assay. Generation of malondialdehyde-like substances was inhibited and delayed by the presence of propolis extracts from group I and II. Our results justify the use of propolis (groups I and II) as a source of natural antioxidants.

Antioxidants↗

Estrogenic effects of ethanol and ether extracts of propolis.

Propolis obtained from honeybee hives has been used in Oriental folk medicine as an anti-inflammatory, anti-carcinogenic, or immunomodulatory agent. The potential estrogenic activity of propolis was investigated in vitro using the MCF-7 human breast cancer cell proliferation, human estrogen receptor (hER) binding and yeast-based steroid receptor transcription, and in vivo using the immature rat uterotrophic effect. Treatments with ethanol extract of propolis (EEP) and ether extract of propolis (REP) enhanced MCF-7 cell proliferation in concentrations ranging from 0.8 to 4 microg/ml. Both EEP and REP competed for binding of [3H]17beta-estradiol to the hER with IC(50) values of 9.14 and 9.72 microg/ml, respectively. In yeast estrogen receptor transcription assay, both EEP and REP were found to be estrogenic with EC(50) values of 9.48, and 8.55 microg/ml, respectively. Animals treated with EEP or REP for 4 days (500-1000 mg/kg per day, s.c.) exhibited significant dose-dependent increases in uterine wet weight. However, in the yeast androgen and progesterone receptor transcription assays, either EEP or REP was found not to be active. The results suggest that propolis produces estrogenic effects through activation of estrogen receptors.

Animals↗

Propolis: lymphocyte proliferation and IFN-gamma production.

We evaluated propolis influence on polyclonal activation of lymphocytes by concanavalin A (Con A). The in vitro experiments showed that propolis decreases splenocyte proliferation both in the absence or presence of Con A. The suppression in mitogen-induced splenocyte proliferation also occurred when mice were treated intraperitoneally with propolis for 3 days. An increased of IFN-gamma production in the culture supernatants of the same cells was observed. A dual action of propolis on lymphocyte activation was proposed: it decreases splenocyte proliferation in the presence or absence of Con A and stimulates IFN-gamma production by spleen cells. These results are important to understand the immunomodulatory action of propolis on the host's specific and non-specific immunity.

Animals↗

Antimicrobial action of propolis and some of its components: the effects on growth, membrane potential and motility of bacteria.

The effect of the natural bee product propolis on the physiology of microorganisms was investigated using B. subtilis, E. coli and R. sphaeroides. An ethanolic extract of propolis had a bactericidal effect caused by the presence of very active, but labile, ingredients. The exact bactericidal effect of propolis was species dependent: it was effective against gram-positive and some gram-negative bacteria. Propolis and some of its cinnamic and flavonoid components were found to uncouple the energy transducing cytoplasmic membrane and to inhibit bacterial motility. These effects on the bioenergetic status of the membrane may contribute to the antimicrobial action of propolis and its observed synergism with selected antibiotics.

Anti-Bacterial Agents↗

Studies of the constituents of Uruguayan propolis.

Eighteen flavonoids including two new compounds, four aromatic carboxylic acids, and eleven phenolic acid esters including one new compound were isolated and identified from the ethyl acetate soluble fraction of the 70% ethanol extract of Uruguayan propolis. The new compounds were elucidated as pinobanksin 3-(2-methyl)butyrate (1; recently reported in Usia, T.; Banskota, A. H.; Tezuka, Y.; Midorikawa, K.; Matsushige, K.; Kadota, S. J. Nat. Prod. 2002, 65, 673-676) pinobanksin 3-isobutyrate (2), and 2-methyl-2-butenyl ferulate (24). The constituents isolated from Uruguayan propolis in this study were similar to those of propolis of European and Chinese origin. Thus, it is suggested that the Uruguayan propolis has a plant origin similar to those of propolis from Europe and China.

Acetates↗

Suppressive effects of ethanolic extracts from propolis and its main botanical origin on dioxin toxicity.

Suppressive effects of ethanolic extracts prepared from propolis group 12 and its main botanical origin (leaf bud of Baccharis dracunculifolia) on transformation of the aryl hydrocarbon receptor (AhR), the initial action of dioxin toxicity, were investigated. It was found that suppressive effects of propolis on AhR transformation were relatively higher than those of resins of its botanical origin in cell-free system and in Hepa-1c1c7 cells. When the composition of chemical ingredients was measured, propolis contained slightly higher amounts of flavonoid aglycones as compared with its botanical origin with the same characteristics. Moreover, antiradical activity, one of the typical biological activities of flavonoids, in propolis was also slightly higher than that in its botanical origin. These results indicate that not only propolis but also its botanical origin contains high amounts of flavonoid aglycones and that both of them are useful dietary sources for flavonoids with a potency to prevent dioxin toxicity.

Animals↗

Renal carcinogenesis induced by ferric nitrilotriacetate in mice, and protection from it by Brazilian propolis and artepillin C.

The protective effect of Brazilian propolis and its extract Artepillin C against ferric nitrilotriacetate (Fe-NTA)-induced renal lipid peroxidation and carcinogenesis was studied in male ddY mice. Fe-NTA-induced renal lipid peroxidation leads to a high incidence of renal cell carcinoma (RCC) in mice. Administration of propolis by gastric intubation 2 h before or Artepillin C at either the same time, 2 h, or 5 h before the intraperitoneal injection of Fe-NTA (7 mg Fe/kg) effectively inhibited renal lipid peroxidation. This was evaluated from the measurement of renal thiobarbituric acid-reactive substances (TBARS) or histochemical findings of 4-hydroxy-2-nonenal (4-HNE)-modified proteins and 8-hydroxy-2'-deoxyguanosine (8-OHdG). Repeated injection of Fe-NTA (10 mg Fe/kg per day, twice a week for a total of 16 times in 8 weeks) caused subacute nephrotoxicity as revealed by necrosis and pleomorphic large nuclear cells in the renal proximal tubules, and gave rise to RCC 12 months later. A protective effect from carcinogenicity was observed in mice given propolis or Artepillin C. Furthermore, the mice given Fe-NTA only developed multiple cysts composed of precancerous lesions with multilayered and proliferating large atypical cells. Mice treated with propolis and Artepillin C also had cysts, but these were dilated and composed of flat cells. These results suggest that propolis and Artepillin C prevent oxidative renal damage and the carcinogenesis induced by Fe-NTA in mice.

8-Hydroxy-2'-Deoxyguanosine↗

Sensitization to propolis in 1255 children undergoing patch testing.

To investigate the frequency and the features of contact sensitization to propolis in children, we patch tested 1255 consecutive children suspected of allergic contact dermatitis with propolis 20% petrolatum over an 8-year period (1995-2002). Positive responses were observed in 5.9% of patch-tested subjects, with a significantly higher frequency in boys than in girls. The dermatitis was localized mainly on the face, hands and limbs. A significant linear increase in the annual frequency of propolis allergy was noted. Confirming literature data, a significant association with balsam of Peru sensitivity was present in children reacting to propolis, linked to the presence of common constituents in the 2 substances. In conclusion, based on our data, we suggest that propolis should not be used in topical products for children because of its high sensitization rate at the paediatric age.

Allergens↗

Anti-hypertensive effects of Brazilian propolis in spontaneously hypertensive rats.

1. Changes in the cardiovascular parameters of Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) were examined following a 4-week diet of either Brazilian propolis or Eucommia uloides OLIVER (tochu). 2. A 4-week diet of propolis or tochu resulted in significant reductions in systolic blood pressure in SHR but had no effect on WKY. Experiments using aorta isolated from animals fed a diet of propolis or tochu revealed increased acetylcholine-induced relaxation in SHR and no change in acetylcholine-induced relaxation in WKY. Sodium nitroprusside-induced relaxation was unaffected by propolis or tochu in both animal groups. 3. These results suggest that propolis and tochu produce an antihypertensive effect that may be mediated by potentiation of acetylcholine-induced vasodilatation.

Animals↗