Christian Scifo,* Venera Cardile,† Alessandra Russo,* Rosanna Consoli,* Carlo Vancheri,‡ Francesco Capasso,§ Angelo Vanella,* and Marcella Renis*1

Clydebank Source

*Department of Biological Chemistry, Medicinal Chemistry and Molecular Biology, and †Department of Physiological Sciences, Viale Andrea Doria, 6, and ‡Department of Internal and Specialistic Medicine, Via Passo Gravina 187- University of Catania, 95125 Catania, Italy §Department of Experimental Pharmacology, Via D. Montesano 49, University of Naples Federico II, 80131 Naples, Italy

Shengli Abstract

Vegetables and fruit help the prevention and the therapy of several kinds of cancer because they contain micronutrients, a class of substances that have been shown to exhibit chemopreventive and chemotherapeutic activities. In the present study the effects of resveratrol (100 and 200 μM), a phytoalexin found in grapes, and of the ethanolic extract of propolis (50 and 100 μg/ml), a natural honeybee hive product, were tested in androgen-resistant prostate cancer cells (DU145), a cell line resembling the last stage of prostate carcinoma. A comparison between the activity of these micronutrients and vinorelbine bitartrate (Navelbine), a semi- synthetic drug normally used in the therapy of prostate cancer, was conducted. Several biochemical parame- ters were tested, such as cell viability (MTT assay), cell membrane integrity (lactate dehydrogenase release), cell redox status (nitric oxide formation, reactive oxygen species production, reduced glutathione levels), genomic DNA fragmentation (COMET assay) with special attention on the presence of apoptotic DNA damage (TUNEL test), and possible mitochondrial transmembrane potential alteration (∆Ψ). Our results point out the anticancer activity of resveratrol and propolis extract in human prostate cancer, exerting their cytotoxicity through two different types of cell death: necrosis and apoptosis, respectively. The data obtained suggest the possible use of these micronutrients both in alternative to classic chemotherapy, and in combina- tion with very low dosage of vinorelbine (5 μM).

Hui-Ping Lin1,2, Shih Sheng Jiang2,3, Chih-Pin Chuu1,2,4*

Source

1 Institute of Cellular and System Medicine, National Health Research Institutes, Miaoli, Taiwan, 2 Translational Center for Glandular Malignancies, National Health Research Institutes, Miaoli, Taiwan, 3National Institute of Cancer Research, National Health Research Institutes, Miaoli, Taiwan, 4Graduate Program for Aging, China Medical University, Taichung, Taiwan

Abstract

Caffeic acid phenethyl ester (CAPE) treatment suppressed proliferation, colony formation, and cell cycle progression in PC-3 human prostate cancer cells. CAPE decreased protein expression of cyclin D1, cyclin E, SKP2, c-Myc, Akt1, Akt2, Akt3, total Akt, mTOR, Bcl-2, Rb, as well as phosphorylation of Rb, ERK1/2, Akt, mTOR, GSK3a, GSK3b, PDK1; but increased protein expression of KLF6 and p21Cip1. Microarray analysis indicated that pathways involved in cellular movement, cell death, proliferation, and cell cycle were affected by CAPE. Co-treatment of CAPE with chemotherapeutic drugs vinblastine, paclitaxol, and estramustine indicated synergistic suppression effect. CAPE administration may serve as a potential adjuvant therapy for prostate cancer.

Chih-Pin Chuu1,2,3,4,5, Hui-Ping Lin3, Mark F. Ciaccio1,2, John M. Kokontis1, Ronald J. Hause Jr1,2, Richard A. Hiipakka1, Shutsung Liao1, and Richard Baker Jones1,2

Source

Cancer Prev Res 2012;5:788-797. Published online May 3, 2012.

Abstract

Caffeic acid phenethyl ester (CAPE) is a bioactive component derived from honeybee hive propolis. CAPE has been shown to have antimitogenic, anticarcinogenic, and other beneficial medicinal properties. Many of its effects have been shown to be mediated through its inhibition of NF-kB signaling pathways. We took a systematic approach to uncover the effects of CAPE from hours to days on the signaling networks in human prostate cancer cells. We observed that CAPE dosage dependently suppressed the proliferation of LNCaP, DU-145, and PC-3 human prostate cancer cells. Administration of CAPE by gavage significantly inhibited the tumor growth of LNCaP xenografts in nude mice. Using LNCaP cells as a model system, we examined the effect of CAPE on gene expression, protein signaling, and transcriptional regulatory networks using micro-Western arrays and PCR arrays. We built a model of the impact of CAPE on cell signaling which suggested that it acted through inhibition of Akt-related protein signaling networks. Overexpression of Akt1 or c-Myc, a downstream target of Akt signaling, significantly blocked the antiproliferative effects of CAPE. In summary, our results suggest that CAPE administration may be useful as an adjuvant therapy for prostate and potentially other types of cancers that are driven by the p70S6K and Akt signaling networks. Cancer Prev Res; 5(5); 788–97. !2012 AACR.

Ikukatsu Suzuki,1 Ikuo Hayashi,1 Takayuki Takaki,1 Debra S. Groveman,2 and Yoshiaki Fujimiya3,4

1Department of Clinical Nutrition, Faculty of Health Sciences, Suzuka University of Medical Science, Suzuka, Mie 510-0293, Japan; 2Veterans Administration Hospital, White River Junction, VT 05009, U.S.A.; 3Department of Basic Medical Sciences, IOND University, Tokyo 165-0027, Japan; and 4R&D Department, Nimura Genetic Solutions, Tokyo 151-051, Japan.

Abstract

Using an ICR mouse model bearing a syngeneic Ehrlich ascitis carcinoma, the present study was under- taken to examine the effects of crude, water-soluble propolis (CWSP) on tumor progression, chemother- apeutic efficacy, and hematopoiesis in the peripheral blood. It was demonstrated that CWSP, adminis- tered subcutaneously, resulted in marked regression of tumor growth in mice, at the early phase after tumor inoculation (CWSP, p , 0.05 vs. saline control). Molecular analysis indicated that the CWSP is composed of 8.4% protein, 4.2% quercetin plus a variety of saccharides with a molecular weight of 29 kDa. Orally administered CWSP did not produce any regression for the observation period (oral CWSP, p . 0.05 vs. saline control). Peritoneal injection of CWSP into neonatal mice resulted in an increased lymphocyte/polymorphonuclear leukocyte ratio activity, indicating the potential activation of lymphoid cell lineages. These observations suggest that subcutaneously injected CWSP could regulate the devel- opment of tumors by possibly stimulating multicellular immunity. In addition, oral administration of CWSP concurrently with 5-fluorouracil (5-FU) or mitomycin C (MMC), significantly increased tumor regression as compared with the respective chemotherapy alone, illustrating the adjuvant effect of orally adminis- tered CWSP for tumor regression when combined with chemotherapeutic agents.
To examine further the potential usefulness of CWSP for chemotherapeutic regimens, which induce pro- found multilineage hematopoietic suppression, mice that received CWSP orally in addition to a 5-FU or MMC were followed for absolute numbers of platelets and white and red blood cells. The oral adminis- tration of CWSP significantly ameliorated the cytopenia induced by 5-FU, resulting in recovery of white as well as red blood cell counts (5-FU plus CWSP, p , 0.05 vs. 5-FU alone or water control; white blood cells on day 15, red blood cells on day 25), but no marked effects on platelet counts was observed (5-FU plus CWSP, p . 0.05 vs. 5-FU alone or water control). On the other hand, CWSP significantly reduced all three MMC-induced cytopenias, especially at the later stage of the chemotherapeutic course (after day 30), suggesting repetitive requirements of oral administration of CWSP.
In summary, subcutaneous administration of an aqueous CWSP resulted in marked regression of trans- planted tumors. Orally administered CWSP combined with chemotherapeutic agents significantly in- creased tumor regression and ameliorated the cytopenia induced by the chemotherapeutic agents alone. These results suggest the benefits of potential clinical trials using CWSP combined with chemotherapeu- tic agents in order to maximize enhanced immunity while potentially minimizing postchemotherapeutic deteriorated reactions.

Nada Oršolic ́ *, Ivan Bašic

Department of Animal Physiology, Faculty of Science, University of Zagreb, Rooseveltov trg 6, 10 000 Zagreb, Croatia

Abstract

Several studies suggest that dietary supplementation with antioxidant can influence the response to chemotherapy as well as the develop- ment of adverse side effects caused by treatment with chemotherapeutic agents. Using CBA mouse model, we investigated a clinically potential use of a water-soluble derivative of propolis (WSDP) in the treatment of various cytopenias induced by radiation and/or chemo- therapy. Also, the antimetastatic efficiency of WSDP given intraperitoneally alone or in combination with chemotherapeutic agents and their effects on the blood leukocytes count as well as on hematopoiesis were studied. Tumor was a transplantable mammary carcinoma (MCa) of CBA mouse. Metastases in the lung were generated by injecting viable tumor cells intravenously (iv). WSDP (50 or 150 mg/kg) exerted a significant antimetastatic effect (P < 0.001) when given either before or after tumor cell inoculation. In combined treatment WSDP and Epirubicin profoundly inhibited metastasis formation; this synergistic effect is maximal when Epirubicin and WSDP were administrated after tumor cell inoculation. Positive outcome of combined treatment with WSDP and Epirubicin was also found regarding the number of red and white blood cells in peripheral blood while in mice treated with Epirubicin alone the significant drop in all hematological parameters was noticed on day 13 after tumor cell inoculation. Furthermore, when WSDP (50 mg/kg) was given perorally (po) for 20 consecutive days an increased number of exogenous CFUs was found in treated mice. WSDP given either for 20 or 40 days increased cellularity of hematopoietic tissue and the number of leucocytes in peripheral blood; prolonged treatment with WSDP also elevated myeloid and megakaryocytic types of CFUs.
To conclude, these findings indicate that the combination of WSDP with chemotherapeutics could increase the antimetastatic potential of chemotherapeutic agents; these findings suggest the benefits of potential clinical trials using WSDP combined with chemotherapeutic agents in order to maximize their antitumor activity and minimize postchemotherapeutic or radiotherapeutic deteriorated reactions. © 2005 Elsevier SAS. All rights reserved.

Sartori G, Pesarico AP, Pinton S, Dobrachinski F, Roman SS, Pauletto F, Junior LC, Prigol M.

Laboratório de Síntese, Reatividade e Avaliação Farmacológica e Toxicológica de Organocalcogênios, Centro de Ciências Naturais e Exatas, Universidade Federal de Santa Maria, Santa Maria, RS, Brazil.

Abstract

Propolis has been highlighted for its antioxidant, anti-inflammatory and antiviral properties. The purpose of this study was to investigate if brown Brazilian hydroalcoholic propolis extract (HPE) protects against vaginal lesions caused by herpes simplex virus type 2 (HSV-2) in female BALB/c mice. The treatment was divided in 5 days of pre-treatment with HPE [50 mg·kg(-1) , once a day, intragastric (i.g.)], HSV-2 infection [10 µl of a solution 1 × 10(2) plaque-forming unit (PFU·ml(-1) HSV-2), intravaginal inoculation at day 6] and post-treatment with HPE (50 mg·kg(-1) ) for 5 days more. At day 11, the animals were killed, and the in vivo analysis (score of lesions) and ex vivo analysis [haematological and histological evaluation; superoxide dismutase (SOD), catalase (CAT) and myeloperoxidase (MPO) activities; reactive species (RS), tyrosine nitration levels, non-protein thiols (NPSH) and ascorbic acid (AA) levels] were carried out. HPE treatment reduced extravaginal lesions and the histological damage caused by HSV-2 infection in vaginal tissues of animals. HPE was able to decrease RS, tyrosine nitration, AA levels and MPO activity. Also, it protected against the inhibition of CAT activity in vaginal tissues of mice. HPE promoted protective effect on HSV-2 infected animals by acting on inflammatory and oxidative processes, and this effect probably is caused by its antioxidant and anti-inflammatory properties. Copyright © 2011 John Wiley & Sons, Ltd.

Ito J, Chang FR, Wang HK, Park YK, Ikegaki M, Kilgore N, Lee KH.
Source

Natural Products Laboratory, School of Pharmacy, University of North Carolina, NC 27599, USA.
Abstract

A new triterpenoid named melliferone (1), three known triterpenoids, moronic acid (2), anwuweizonic acid (3), and betulonic acid (4), and four known aromatic compounds (5-8) were isolated from Brazilian propolis and tested for anti-HIV activity in H9 lymphocytes. Moronic acid (2) showed significant anti-HIV activity (EC(50) <0.1 microg/mL, TI >186) and was modified to develop more potent anti-AIDS agents.

T. F. Bachiega, C. L. Orsatti, A. C. Pagliarone and J. M. Sforcin*

Department of Microbiology and Immunology, Biosciences Institute, UNESP, Botucatu, SP, 18618-970, Brazil

Abstract

Since propolis and phenolic compounds, such as cinnamic and coumaric acids, have several biological properties, their immunomodulatory effect on cytokine production (IL-1b, IL-6 and IL-10) was investigated. Peritoneal macrophages from BALB/c mice were incubated with propolis, coumaric and cinnamic acids in different concentrations and the concentrations that inhibited cytokine production were tested before or after macrophage challenge with LPS, to evaluate a possible immunomodulatory action. Propolis and the acids stimulated IL-1b production, while IL-6 production was signi␣cantly inhibited after incubation with propolis (5, 50 and 100mg/well), coumaric and cinnamic acids (50 and 100mg/well). In LPS-challenge protocols, inhibitory concentrations of cinnamic and coumaric acids after LPS incubation prevented ef␣ciently its effects on IL-6 production, whereas propolis inhibited LPS effects both before and after its addition. Propolis, coumaric and cinnamic acids (50 and 100 mg/well) inhibited IL-10 production as well. Both acids showed a similar inhibitory activity on IL-10 production when added after LPS challenge, while propolis counteracted LPS action when added before and after LPS incubation. Propolis modulated the immune/in␣ammatory response, depending on the concentration. Its ef␣ciency may occur due to the synergistic effect of its compounds, and cinnamic and coumaric acids may be involved in the action of propolis on cytokine production. Copyright © 2012 John Wiley & Sons, Ltd.

Shimizu T, Hino A, Tsutsumi A, Park YK, Watanabe W, Kurokawa M.
Source

Department of Biochemistry, School of Pharmaceutical Sciences, Kyushu University of Health and Welfare, Nobeoka, Miyazaki, Japan.
Abstract

BACKGROUND:

Propolis has been used worldwide as a dietary supplement to maintain and improve human health. We examined whether ethanol extracts of Brazilian propolis exhibit antiviral activity against influenza virus in vitro and in vivo.
METHODS:

Among 13 ethanol extracts screened in a plaque reduction assay, four showed anti-influenza virus activity. The anti-influenza efficacy of the four extracts was further examined in a murine influenza virus infection model. The mice were infected intranasally with influenza virus, and the four extracts were orally administered at 10 mg/kg three times daily for seven successive days after infection.
RESULTS:

In this infection model, only one extract, AF-08, was significantly effective at 10 mg/kg in reducing the body weight loss of infected mice. The doses of 2 and 10 mg/kg were also effective in prolonging the survival times of infected mice significantly, but 0.4 mg/kg was not. The anti-influenza efficacy of AF-08 at 10 mg/kg was confirmed in a dose-dependent manner in mice. AF-08 at 10 mg/kg significantly reduced virus yields in the bronchoalveolar lavage fluids of lungs in infected mice as compared with the control. The reduction of virus yields by AF-08 at 10 mg/kg significantly corresponded to those induced by oseltamivir at 1 mg/kg twice daily from day 1 to day 4 after infection.

CONCLUSION:

The Brazilian propolis AF-08 was indicated to possess anti-influenza virus activity and to ameliorate influenza symptoms in mice. AF-08 may be a possible candidate for an anti-influenza dietary supplement for humans.

Basnet P, Matsushige K, Hase K, Kadota S, Namba T.
Source

Research Institute for Wakan-Yaku (Traditional Sino-Japanese Medicines), Toyama Medical and Pharmaceutical University, Japan.

Abstract

The water extract of propolis (PWE) showed a strong hepatoprotective activity against CCl4-toxicity in rats and D-galactosamine (GalN)/lipopolysaccharide (LPS)-induced liver injury in mice. The PWE also showed a significant hepatoprotective activity against CCl4-induced liver cell injury in cultured rat hepatocytes. The in vitro hepatoprotective activity guided fractionation and chemical analysis led to the isolation of four dicaffeoyl quinic acid derivatives from the PWE. The structure of these isolates was determined to be methyl 3,4-di-O-caffeoyl quinate (1), 3,4-di-O-caffeoyl quinic acid (2), methyl 4,5-di-O-caffeoyl quinate (3), and 3,5-di-O-caffeoyl quinic acid (4) by spectroscopic methods. These compounds were more potent hepatoprotective agents than glycyrrhizin at a concentration of 10 micrograms/ml and 1 was the most potent among the four compounds in the cultured hepatocytes. Quinic acid (5) alone did not show hepatoprotective effects in cultured rat hepatocytes against CCl4-toxicity. On the other hand, chlorogenic acid (6) or caffeic acid alone was found to be less potent than the dicaffeoyl quinic acid derivatives.

Basnet P, Matsushige K, Hase K, Kadota S, Namba T.
Source

Research Institute for Wakan-Yaku (Traditional Sion-Japenese Medicines), Toyama Medicine and Pharmaceutical University, Japan.
Abstract

Hepatoprotective activity guided chemical analyses led to the isolation of two dicaffeoyl quinic acid derivatives, methyl 3,4-di-O-caffeoyl quinate (1) and 3,4-di-O-caffeoyl quinic acid (2) from water extract of propolis, and their structures were determined by the use of 2D NMR. These compounds were stronger antihepatotoxic agents than glycyrrhizin.

Banskota AH, Tezuka Y, Adnyana IK, Midorikawa K, Matsushige K, Message D, Huertas AA, Kadota S.
Source

Department of Natural Products Chemistry, Institute of Natural Medicine, Toyama Medical and Pharmaceutical University, 2630-Sugitani, Toyama 930-0194, Japan.

Abstract

Propolis is a resinous hive product collected by honeybees from various plant sources. The composition of the propolis depends upon the time, vegetation and the area of collection. Thus, quality evaluation of the propolis is important, before use in food and beverages. For this propose three different biological activities were carried out, i.e. 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, cytotoxicity and hepatoprotective activity, of MeOH and water extracts of nine different propolis from Brazil, Peru, the Netherlands and China. The results showed that water extracts of six Brazilian and a Chinese propolis possessed stronger DPPH free radical scavenging activity than the corresponding MeOH extract, whereas in the case of Netherlands and Peruvian propolis MeOH extract exhibited stronger DPPH free radical scavenging activity. The MeOH extracts of all propolis possessed stronger cytotoxicity than the corresponding water extract towards murine colon 26-L5 carcinoma and human HT-1080 fibrosarcoma cells. The result of hepatoprotective activity of Brazilian propolis on D-galactosamine (D-GalN)/tumor necrosis factor-alpha (TNF-alpha)-induced cell death in primary cultured mouse hepatocytes were found in accordance with the grade set up by beekeepers in Brazil.

Park YK, Koo MH, Abreu JA, Ikegaki M, Cury JA, Rosalen PL.
Source

College of Food Engineering, State University of Campinas (UNICAMP), 13081-970, Caixa Postal 6177, Campinas, SP, Brazil.

Abstract

Formation of dental caries is caused by the colonization and accumulation of oral microorganisms and extracellular polysaccharides that are synthesized from sucrose by glucosyltransferase of Streptococcus mutans. The production of glucosyltransferase from oral microorganisms was attempted, and it was found that Streptococcus mutans produced highest activity of the enzyme. Ethanolic extracts of propolis (EEP) were examined whether EEP inhibit the enzyme activity and growth of the bacteria or not. All EEP from various regions in Brazil inhibited both glucosyltransferase activity and growth of S. mutans, but one of the propolis from Rio Grande do Sul (RS2) demonstrated the highest inhibition of the enzyme activity and growth of the bacteria. It was also found that propolis (RS2) contained the highest concentrations of pinocembrin and galangin.

Castro ML, do Nascimento AM, Ikegaki M, Costa-Neto CM, Alencar SM, Rosalen PL.
Source

Department of Physiological Sciences, Piracicaba Dental School, University of Campinas (UNICAMP), C.P. 52, 13414-903, Piracicaba, SP, Brazil.
Abstract

A prenylated benzophenone, hyperibone A, was isolated from the hexane fraction of Brazilian propolis type 6. Its structure was determined by spectral analysis including 2D NMR. This compound exhibited cytotoxic activity against HeLa tumor cells (IC(50)=0.1756microM), strong antimicrobial activity (MIC range-0.73-6.6microg/mL; MBC range-2.92-106microg/mL) against Streptococcus mutans, Streptococcus sobrinus, Streptococcus oralis, Staphylococcus aureus, and Actinomyces naeslundii, and the results of its cytotoxic and antimicrobial activities were considered good.

Banskota AH, Tezuka Y, Midorikawa K, Matsushige K, Kadota S.
Source

Institute of Natural Medicine, Toyama Medical and Pharmaceutical University, 2630-Sugitani, Toyama 930-0194, Japan.
Abstract

Two novel benzofuran derivatives, propolis-benzofurans A (1) and B (2), were isolated from the MeOH extract of Brazilian propolis, together with two known isoprenylated compounds (E)-3-[2, 3-dihydro-2-(1-methylethenyl)-7-prenyl-5-benzofuranyl]-2-propen oic acid and (E)-3-¿4-hydroxy-3-[(E)-4-(2, 3-dihydrocinnamoyloxy)-3-methyl-2-butenyl]-5-prenylphenyl¿-2 -propenoi c acid. The structures of these compounds were elucidated on the basis of spectral analysis. Both the new compounds exhibited mild cytotoxicity toward highly liver-metastatic murine colon 26-L5 carcinoma and human HT-1080 fibrosarcoma cells.

Recent progress in pharmacological research of propolis.
Banskota AH, Tezuka Y, Kadota S.
Source

Department of Natural Products Chemistry, Institute of Natural Medicine, Toyama Medical and Pharmaceutical University, 2630-Sugitani, Toyama 930-0194, Japan.

Abstract

Propolis is a resinous hive product collected by honeybees from various plant sources. It is a popular folk medicine possessing a broad spectrum of biological activities. It has also been used as a health drink in various Asian, European and American countries. Several groups of researchers have focused their attention on the biological activity of propolis and its active principles. Many scientific articles are published every year in different international journals related to the pharmacological properties of propolis. This review article compiles recent findings (since 1995) on the pharmacological properties of propolis focusing on its antihepatotoxic, antitumour, antioxidative, antimicrobial and antiinflammatory properties. The possible mechanism of action of propolis as well as the active compounds are discussed.

Akao Y, Maruyama H, Matsumoto K, Ohguchi K, Nishizawa K, Sakamoto T, Araki Y, Mishima S, Nozawa Y.
Source

Gifu International Institute of Biotechnology, Kakamigahara, Japan. yakao@giib.or.jp
Abstract

A cell growth inhibitory effect of drupanin and baccharin, ingredients of propolis, was found in human cancer cell lines. These compounds induced apoptosis in the cells characterized by morphological and nucleosomal DNA fragmentation analysis. Their effects were less potent compared with that of artepillin C, which is a known anticancer compound from propolis. Importantly, HL60 cells were more sensitive to drupanin than were Con A-stimulated peripheral blood lymphocytes, whereas the potency of artepillin C was the opposite of that of drupanin.

Mishima S, Ono Y, Araki Y, Akao Y, Nozawa Y.
Source

API Co. Ltd., R&D, Gifu, Japan.
Abstract

Honey bee propolis is rich in cinnamic acid derivatives. Baccharin and drupanin from Brazilian honey bee propolis are cinnamic acid derivatives that contain prenyl moieties. We previously isolated these two compounds and demonstrated that they induce an apoptotic event in several tumor cell lines. In this study, we examined the tumoricidal activity of baccharin and drupanin in mice allografted with sarcoma S-180 and also studied the genotoxic effects on normal splenocytes using the alkaline single cell gel (comet) assay. We found that both baccharin and drupanin effectively suppressed growth of the tumor. Furthermore, these compounds induced a significant genotoxic effect on the tumor cells in comparison with normal splenocytes. Thus, baccharin and drupanin are potent tumor suppressive components of honeybee propolis.

Mishima S, Inoh Y, Narita Y, Ohta S, Sakamoto T, Araki Y, Suzuki KM, Akao Y, Nozawa Y.
Source

Nagaragawa Research Center, API Co., Ltd., 692-3 Nagara, Gifu 502-0071, Japan.
Abstract

We have previously reported that Brazilian propolis extracts inhibited growth of HL-60 human myeloid leukemia cells, which is partly attributed to the induction of apoptosis associated with granulocytic differentiation. In this study, we isolated three compounds which induce granulocytic differentiation evaluated by nitroblue tetrazolium (NBT)-reducing assays from the water extract of propolis and identified as 4,5-di-O-caffeoylquinic, 3,5-di-O-caffeoylquinic, and 3,4-di-O-caffeoylquinic acids by NMR analysis. Cell growth inhibitory activity of these caffeoylquinic acids was found in HL-60 cell, which was mainly attributed to the induction of apoptosis. Furthermore, the potency of caffeoylquinic acid derivatives to induce granulocytic differentiation was examined in HL-60 cells. Caffeic, quinic, and chlorogenic acids had no effects on the NBT-reducing activity, while 3,4,5-tri-O-caffeoylquinic acid induced more than 30% of NBT-positive cells. These results suggest that the number of the caffeoyl groups bound to quinic acid plays an important role in the potency of the caffeoylquinic acid derivatives to induce granulocytic differentiation. This is the first report demonstrating that the caffeoylquinic acid derivatives induce granulocytic differentiation of HL-60 cells.

Koo H, Rosalen PL, Cury JA, Ambrosano GM, Murata RM, Yatsuda R, Ikegaki M, Alencar SM, Park YK.
Source

Department of Physiological Sciences, Faculty of Dentistry of Piracicaba, State University of Campinas, Caixa Postal 52, Piracicaba, 13414-900, SP, Brazil. Hyun-Koo@urmc.rochester.edu
Abstract

The effects of a new variety of propolis, from Northeastern Brazil (BA), on growth of mutans streptococci, cell adherence, and water-insoluble glucan (WIG) synthesis were evaluated. Propolis from Southeastern (MG) and Southern (RS) Brazil were also tested as an extension of our previous work. Ethanolic extracts of propolis (EEP) were prepared and analyzed by reversed-phase HPLC. For the antibacterial activity assays, minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of EEPs against Streptococcus mutans, S. sobrinus, and S. cricetus were determined. Cell adherence of S. mutans and S. sobrinus to a glass surface was measured spectrophotometrically at 550 nm. WIG synthesized from sucrose by glucosyltransferase (Gtf) was extracted and quantified by the phenol-sulfuric method. The HPLC profile of the new variety of propolis was entirely different from Southeastern and Southern propolis. Neither flavonoid aglycones nor p-coumaric acid were detected in EEP BA. All EEPs demonstrated biological activities against mutans streptococci; EEP BA showed the highest potency in all in vitro parameters evaluated in this study. The ranges of MIC values were 50 (EEP BA)-400 microg/ml (MG), for S. mutans; and 25 (BA)-400 microg/ml (MG), for S. sobrinus and S. cricetus. The bactericidal concentration of EEPs was four to eight times the MIC values. The adherence of S. mutans and S. sobrinus cells and WIG synthesis were markedly inhibited by EEPs, demonstrating significant inhibition at all concentrations compared with the control (80% ethanol) (p<0.05). EEP BA showed 80% inhibition of cell adherence and WIG synthesis at concentrations as low as 12.5 and 7.8 microg/ml, respectively. The results show that the new variety of propolis was exceptionally effective in all in vitro parameters tested against mutans streptococci; biological effects of propolis are likely not to be due solely to flavonoids and (hydroxy)cinnamic acid derivatives.

Park YK, Koo MH, Abreu JA, Ikegaki M, Cury JA, Rosalen PL.
Source

College of Food Engineering, State University of Campinas (UNICAMP), 13081-970, Caixa Postal 6177, Campinas, SP, Brazil.
Abstract

Formation of dental caries is caused by the colonization and accumulation of oral microorganisms and extracellular polysaccharides that are synthesized from sucrose by glucosyltransferase of Streptococcus mutans. The production of glucosyltransferase from oral microorganisms was attempted, and it was found that Streptococcus mutans produced highest activity of the enzyme. Ethanolic extracts of propolis (EEP) were examined whether EEP inhibit the enzyme activity and growth of the bacteria or not. All EEP from various regions in Brazil inhibited both glucosyltransferase activity and growth of S. mutans, but one of the propolis from Rio Grande do Sul (RS2) demonstrated the highest inhibition of the enzyme activity and growth of the bacteria. It was also found that propolis (RS2) contained the highest concentrations of pinocembrin and galangin.

Hayacibara MF, Koo H, Rosalen PL, Duarte S, Franco EM, Bowen WH, Ikegaki M, Cury JA.
Source

Department of Physiological Sciences, Dentistry Faculty of Piracicaba, State University of Campinas, Av. Limeira, 901 PIRACICABA-SP, CEP: 13414-903, Brazil. mhayacibara@wnet.com.br
Abstract

Recently, two chemically different types of Brazilian propolis (type-3 and -12) were shown to have cariostatic properties. This study aimed to evaluate the influence of their isolated fractions on mutans streptococci viability, glucosyltransferases (GTFs) activity and caries development in rats. The ethanolic extracts of propolis (EEPs) were serially fractionated into hexane (H-fr), chloroform, ethyl acetate, and ethanol. The ability of the four fractions and EEP to inhibit Streptococcus mutans and Streptococcus sobrinus growth and adherence to a glass surface was examined. The effect on GTFs B and C activity was also determined. For the caries study, 60 Wistar rats infected with Streptococcus sobrinus were treated topically twice daily as follows: (1) EEP type-3, (2) H-fr type-3, (3) EEP type-12, (4) H-fr type-12, and (5) control. In general, the H-fr from both types of propolis showed the highest antibacterial activity and GTFs inhibition. Furthermore, the EEP and H-fr type-3 and -12 were equally effective in reducing dental caries in rats. The data suggest that the putative cariostatic compounds of propolis type-3 and -12 are mostly non-polar; and H-fr should be the fraction of choice for identifying further potentially novel anti-caries agents.

Quincoces Suarez JA, Rando DG, Santos RP, Gonçalves CP, Ferreira E, de Carvalho JE, Kohn L, Maria DA, Faião-Flores F, Michalik D, Marcucci MC, Vogel C.
Source

Laboratory of Organic Synthesis, Universidade Bandeirante de São Paulo (UNIBAN), São Paulo, Brazil. quinco99@hotmail.com
Abstract

This paper describes a new method for the preparation of 1,5-bis(4-hydroxy-3-methoxyphenyl)-1,4-pentadien-3-one 1 and its derivatives 2-5. This set of synthetic compounds exhibited high antitumoral activities regarding in vitro screening against several human tumor cell lines as lung carcinoma NCI-460, melanoma UACC-62, breast MCF-7, colon HT-29, renal 786-O, ovarian OVCAR-03 and ovarian expressing the resistance phenotype for adriamycin NCI-ADR/RES, prostate PC-3, and leukemia K-562. Compounds were also tested against murine tumor cell line B16F10 melanoma and lymphocytic leukemia L1210 as well as to their effect toward normal macrophages. Specific activity against colon cancer cells HT-29 was observed for all tested compounds and suggests further studies with models of colon cancer. Compounds 1, 2, and 4 showed significant cytotoxic activity with IC(50) values 2.3 microM for all human cancer cell lines. Intraperitoneal acute administration of compound 1 and 2 showed very low toxicity rate.

Copyright 2010 Elsevier Ltd. All rights reserved.

Marcucci MC, Ferreres F, García-Viguera C, Bankova VS, De Castro SL, Dantas AP, Valente PH, Paulino N.
Source

Laboratorio de Fitoquímica, CEBAS (CSIC) Apdo Correos 4195, 30080, Murcia, Spain. mcmarcucci@hotmail.com
Abstract

Four compounds were isolated from Brazilian propolis. They are identified as: (1) 3-prenyl-4-hydroxycinnamic acid (PHCA), (2) 2,2-dimethyl-6-carboxyethenyl-2H-1-benzopyrane (DCBEN), (3) 3,5-diprenyl-4-hydroxycinnamic acid (DHCA), and (4) 2,2-dimethyl-6-carboxyethenyl-8-prenyl-2H-1-benzopyran (DPB). The structures of the compounds were determined by MS and NMR techniques. All compounds were assayed against Trypanosoma cruzi and the bacteria Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Streptococcus faecalis. Compounds (1) to (4) were active against T. cruzi. Except (1), all compounds presented activity against the bacteria tested. When compounds (1)-(3) were tested in the guinea pig isolated trachea, all induced a relaxant effect similar to propolis extract.

Pereira RM, Andrades NE, Paulino N, Sawaya AC, Eberlin MN, Marcucci MC, Favero GM, Novak EM, Bydlowski SP.
Source

Department of Pharmacy, Bandeirante University of São Paulo, Rua Maria Cândida 1813, São Paulo, SP, Brazil. rpereira02@hotmail.com
Abstract

The antioxidant activity of flavonoids is believed to increase when they are coordinated with transition metal ions. However, the literature on this subject is contradictory and the outcome seems to largely depend on the experimental conditions. In order to understand the contribution of the metal coordination and the type of interaction between a flavonoid and the metal ion, in this study a new metal complex of Cu (II) with naringin was synthesized and characterized by FT-IR, UV-VIS, mass spectrometry (ESI-MS/MS), elemental analysis and 1H-NMR. The results of these analyses indicate that the complex has a Cu (II) ion coordinated via positions 4 and 5 of the flavonoid. The antioxidant, anti-inflammatory and antimicrobial activities of this complex were studied and compared with the activity of free naringin. The Naringin-Cu (II) complex 1 showed higher antioxidant, anti-inflammatory and tumor cell cytotoxicity activities than free naringin without reducing cell viability.

A. Montoro,1* J. F. Barquinero,2 M. Almonacid,1 A. Montoro,3 N. Sebastià,3 G. Verdú,4 V. Sahuquillo,1 J. Serrano,5 M. Saiz,5 J. I. Villaescusa,1 and J. M. Soriano3
1Servicio de Protección Radiológica, Hospital Universitario La Fe, 46009 Valencia, Spain
2Unitat d’Antropologia Biològica, Departamento de Biologia Animal, Biologia Vegetal i Ecologia, Facultat de Biociències, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain
3Área de Nutrición y Bromatología, Facultat de Farmàcia, Universitat de València, 46100 Burjassot, Spain
4Departamento de Ingeniería Química y Nuclear, Escuela Superior de Ingenieros Industriales, Universidad Politécnica de Valencia, 46022 Valencia, Spain
5Dietéticos Intersa, Plaza Dr. Seres no. 13, Torreserona, Lleida 25131, Spain
*A. Montoro: Email: montoro_ale@gva.es

Abstract

Radioprotection with natural products may be relevant to the mitigation of ionizing radiation-induced damage in mammalian systems; in this sense, propolis extracts have shown effects such as antioxidant, antitumoral, anti-inflammatory, and immunostimulant. We report for the first time a cytogenetic study to evaluate the radioprotective effect, in vitro, of propolis against radiation-induced chromosomal damage. Lymphocytes were cultured with increasing concentrations of ethanol extract of propolis (EEP), including 20, 40, 120, 250, 500, 750, 1000, and 2000 μg mL−1 and then exposed to 2 Gy γ-rays. A significant and concentration-dependent decrease is observed in the frequency of chromosome aberrations in samples treated with EEP. The protection against the formation of dicentrics was concentration-dependent, with a maximum protection at 120 μg mL−1 of EEP. The observed frequency of dicentrics is described as negative exponential function, indicating that the maximum protectible fraction of dicentrics is approximately 44%. Free radical scavenging and antioxidant activities are the mechanisms that these substances use to protect cells from ionizing radiation.

Kelen Fátima Dalben Dota,1 Marcia Edilaine Lopes Consolaro,1 Terezinha Inez Estivalet Svidzinski,1 and Marcos Luciano Bruschi2*
1Graduate Program of Health Sciences, State University of Maringa, Parana, Brazil
2Department of Pharmacy, State University of Maringa, Colombo Avenue, 5790, CEP 87020-900, Maringa, Parana, Brazil
*Marcos Luciano Bruschi: Email: mlbruschi@uem.br

Abstract
Propolis, a resinous compound produced by Apis mellifera L. bees, is known to possess a variety of biological activities and is applied in the therapy of various infectious diseases. The aim of this study was to evaluate the in vitro antifungal activity of propolis ethanol extract (PE) and propolis microparticles (PMs) obtained from a sample of Brazilian propolis against clinical yeast isolates of importance in the vulvovaginal candidiasis (VVC). PE was used to prepare the microparticles. Yeast isolates (n = 89), obtained from vaginal exudates of patients with VVC, were exposed to the PE and the PMs. Moreover, the main antifungal drugs used in the treatment of VVC (Fluconazole, Voriconazole, Itraconazole, Ketoconazole, Miconazole and Amphotericin B) were also tested. Minimum inhibitory concentration (MIC) was determined according to the standard broth microdilution method. Some Candida albicans isolates showed resistance or dose-dependent susceptibility for the azolic drugs and Amphotericin B. Non-C. albicans isolates showed more resistance and dose-dependent susceptibility for the azolic drugs than C. albicans. However, all of them were sensitive or dose-dependent susceptible for Amphotericin B. All yeasts were inhibited by PE and PMs, with small variation, independent of the species of yeast. The overall results provided important information for the potential application of PMs in the therapy of VVC and the possible prevention of the occurrence of new symptomatic episodes.