Ethanol Extract Of Brazilian Propolis Research Articles

Flow cytometric detection of CD11b+ Gr-1+ cells in nontumor-bearing mice: A propolis-elicited model.

Myeloid-derived suppressor cells (MDSCs) are a heterogenous myeloid lineage population whose conventional surface phenotype is CD11b+ Gr-1+. Due to their rarity and fragility, analyses using primary isolated MDSCs are extremely difficult. However, counting CD11b+ Gr-1+ cells in associated tissues such as tumors and inflammatory lesions provides critical information regarding MDSC involvement in immune disorders in the tissues. Specific MDSC markers have not been identified, limiting our ability to apply histochemical approaches during MDSCs research. However, profiling surface antigens using multi-colorimetric flow cytometry enables us to easily monitor the abundance of MDSCs in vivo. Monitoring of mouse MDSCs and their subpopulations using flow cytometry is well established. In this article, I exemplify a conventional method of monitoring CD11b+ Gr-1+ cells in mouse adipose tissue after administration of Brazilian propolis ethanol extract, which is a strong inducer of MDSCs.

Brazilian propolis extract reduces intestinal barrier defects and inflammation in a colitic mouse model

Brazilian propolis is rich in cinnamic acid derivatives and reportedly reduces intestinal inflammation in rodents; however, the underlying mechanisms remain unclear. We hypothesized that the regulation of tight junction (TJ) barrier, Th17 cell differentiation, and/or, macrophage activation by cinnamic acid derivatives were involved in the propolis-mediated anti-inflammatory effect. Mice were orally administered 2% dextran sodium sulfate (DSS) in combination with either the feeding control or a diet containing 0.3% ethanol extract of Brazilian propolis for 9 days. DSS administration induced acute colitis in mice, whereas the propolis extract mitigated DSS-induced weight loss; colon shortening; increased plasma levels of lipopolysaccharide-binding protein; reduced expression of TJ proteins, such as zonula occludens, junctional adhesion molecule-A, occludin, and claudins; and increased expression of inflammatory cytokines, such as tumor necrosis factor (TNF) α, interleukin (IL) 6, and IL-17a. Cinnamic acid derivatives, such as artepillin C and caffeic acid phenethyl ester, present in the propolis extract suppressed the IL-17 production from cultured murine splenocytes through decreased retinoic acid-related orphan receptor gT expression. Baccharin, drupanin, and culifolin, which are also present in Brazilian propolis, reduced the TNF-α and/or IL-6 production by suppressing inflammatory signaling in murine RAW 264.7 macrophages. Taken together, the regulation of Th17 differentiation and macrophage activation by cinnamic acid derivatives, at least in part, contribute to the anti-inflammatory effect mediated by Brazilian propolis.

Antioxidant activity and lipid oxidation in milk from cows with soybean oil and propolis extract added to their feed

Abstract The presence of PUFA (polyunsaturated fatty acids) in cow milk, including conjugated linoleic acids (CLA) isomers, makes milk susceptible to lipid oxidation, and may lead to the formation of volatile products responsible for a rancid smell. Consequently, including natural antioxidants in the diet of dairy cattle can assist in preventing lipid oxidation. In this study, soybean oil (a source of PUFA) and different ethanolic extracts of Brazilian propolis were included in the cows' feed, and the antioxidant activity and lipid oxidation of the animals' milk was assessed. Antioxidant activity was analyzed using the DPPH method, potassium ferricyanide reduction and chelating ability. Total phenolic compounds and flavonoids were also determined. Lipid oxidation was assessed by determining conjugated dienes and the production of aldehydes (propanal, pentanal, hexanal and octanal). Increased antioxidant activity was seen in the samples of milk from cows receiving propolis. Similarly, the addition of ethanolic extract of propolis to the diet of dairy cattle increased milk protection for two treatments with propolis against lipid oxidation in terms of production of propanal and hexanal. These results confirm the positive effect of applying this natural additive to the diet of lactating cows, improving the nutritional quality of the milk produced.

Brazilian propolis ethanol extract and its component kaempferol induce myeloid-derived suppressor cells from macrophages of mice in vivo and in vitro

BackgroundBrazilian green propolis is produced by mixing secretions from Africanized honey bees with exudate, mainly from Baccharis dracunculifolia. Brazilian propolis is especially rich in flavonoids and cinammic acid derivatives, and it has been widely used in folk medicine owing to its anti-inflammatory, anti-viral, tumoricidal, and analgesic effects. Moreover, it is applied to prevent metabolic disorders, such as type 2 diabetes and arteriosclerosis. Previously, we demonstrated that propolis ethanol extract ameliorated type 2 diabetes in a mouse model through the resolution of adipose tissue inflammation. The aims of this study were to identify the immunosuppressive cells directly elicited by propolis extract and to evaluate the flavonoids that induce such cells.MethodsEthanol extract of Brazilian propolis (PEE; 100 mg/kg i.p., twice a week) was injected into lean or high fat-fed obese C57BL/6 mice or C57BL/6 ob/ob mice for one month. Subsequently, immune cells in visceral adipose tissue and the peritoneal cavity were monitored using FACS analysis. Isolated macrophages and the macrophage-like cell line J774.1 were treated with PEE and its constituent components, and the expression of immune suppressive myeloid markers were evaluated. Finally, we injected one of the identified compounds, kaempferol, into C57BL/6 mice and performed FACS analysis on the adipose tissue.ResultsIntraperitoneal treatment of PEE induces CD11b+, Gr-1+ myeloid-derived suppressor cells (MDSCs) in visceral adipose tissue and the peritoneal cavity of lean and obese mice. PEE directly stimulates cultured M1 macrophages to transdifferentiate into MDSCs. Among twelve compounds isolated from PEE, kaempferol has an exclusive effect on MDSCs induction in vitro. Accordingly, intraperitoneal injection of kaempferol causes accumulation of MDSCs in the visceral adipose tissue of mice.ConclusionBrazilian PEE and its compound kaempferol strongly induce MDSCs in visceral adipose tissue at a relatively early phase of inflammation. Given the strong anti-inflammatory action of MDSCs, the induction of MDSCs by PEE and kaempferol is expected to be useful for anti-diabetic and anti-inflammatory therapies.Graphical

A comparative assessment of effects of Polish propolis combined with plant oils with those of Brazilian propolis on the oral cavity condition in ortofacial cleft patients treated with removable appliances

In patients with orofacial cleft, toothpaste they use should demonstrate anti-inflammatory, antibacterial and regenerative action and at the same time be well-tolerated by the oral cavity epithelium and characterized by low toxicity. For this reason, it is expedient to seek preparations that would show the properties described above and could be used as auxiliary means supporting the maintenance of oral health in patients with orofacial clefts. Aim. The aim of the study was to perform a comparative assessment of toothpaste containing Polish propolis in combination with plant oils with that containing Brazilian propolis on the oral cavity condition in patients with orofacial cleft treated with removable appliances. Material and methods. The study material included 75 patients eligible for the study with a cleft lip or cleft lip and palate treated orthodontically. They were divided into three groups, A, B and C. Group A received a toothpaste with 2 % ethanol extract of Polish propolis (EEP-P) and plant oils, 0.5% tea tree oil (TTO), 0.2% menthol oil (MO), and 0.1% rosemary oil (RO). Group B received a toothpaste with 3% ethanol extract of Brazilian propolis (EEP-B). Group C included patients who were given a placebo toothpaste devoid of any active substances. During the baseline (first) and a follow-up (final) examination after 35 days, in each patient the Approximate Plaque Index (API), gingival index (GI) and Bleeding on Probing (BOP) were assessed. Results. The biggest improvement of the gingival condition translated into the GI index and the BOP was found in group A and amounted to 31% and 26% respectively, compared to the baseline examination (p<0.00); a statistically significant reduction of the GI index by 25% and BOP by 29% (p<0.01) was found in group B. No statistically significant differences of the values of the API, GI and BOP indices were found between groups A and B. There was no significant improvement of the gingiva condition in Group C. Conclusions. Both types of toothpaste containing active substances have beneficial effects on the condition of the marginal parodontium and they do not significantly differ in terms of their biological activity in the oral cavity. Besides, the toothpaste with Polish propolis and plant oils has a beneficial effect on the oral cavity hygiene in children with orofacial cleft. (Machorowska-Pieniążek A, Tanasiewicz M, Skucha-Nowak M, Morawiec T. A comparative assessment of effects of Polish propolis combined with plant oils with those of Brazilian propolis on the oral cavity condition in orofacial cleft patients treated with removable appliances. Orthod Forum 2017; 13: 15-27).

Ethanol extract of Brazilian propolis ameliorates cognitive dysfunction and suppressed protein aggregations caused by hyperhomocysteinemia.

Homocysteine (Hcy) has been proposed to be a risk factor for cognitive dysfunction. We investigated the effects and the underlying mechanisms of action of propolis, which has antioxidant activity on Hcy-induced oxidative stress in vitro and in vivo. For the in vitro assays, neuroblastoma SH-SY5Y and glioblastoma U-251MG cells were cultured with Hcy and various concentrations of propolis. Cell death and reactive oxygen species production were significantly suppressed by propolis in dose-dependent manner, compared with Hcy alone. For the in vivo assays, mice were fed a propolis-containing diet and Hcy thiolactone in water. Cognitive function was evaluated using the Morris water maze test. Propolis suppressed cognitive dysfunction caused by hyperhomocysteinemia. Accumulation of aggregated protein in brain was accelerated in hyperhomocysteinemia, and the accumulation was suppressed by propolis. Hyperhomocysteinemia, however, did not enhance the oxidative stress in brain. In vitro amyloid formation assay showed that Hcy accelerated lysozyme aggregation and propolis inhibited the aggregation.

Oral administration of Brazilian propolis exerts estrogenic effect in ovariectomized rats.

Propolis, a natural product derived from plants by honeybees, is a mixture of several hundred chemicals, including flavonoids, coumaric acids, and caffeic acids, some of which show estrogen-like activity. In this study, the estrogenic activity of crude ethanolic extract of Brazilian propolis was determined using several in vitro and in vivo assays. Propolis was found to bind to human estrogen receptors (ERs). Furthermore, propolis induced the expression of estrogen-responsive genes in ER-positive MCF-7 and Ishikawa cells. These in vitro assays suggest that propolis exerts estrogenic activity; therefore, in vivo experiments were conducted using ovariectomized rats. Oral administration of propolis (55 or 550 mg/kg/day for 3 days) significantly increased uterine wet weight and luminal epithelium thickness in comparison with the corresponding values in the corn oil-treated control group. Moreover, propolis induced ductal cell proliferation in the mammary glands. These effects were completely inhibited by full ER antagonist ICI 182,780, confirming that the effects of propolis are mediated by the ER. Our data show that oral intake of propolis induces estrogenic activity in ER-expressing organs in vivo and suggest that Brazilian propolis is a useful dietary source of phytoestrogens and a promising treatment for postmenopausal symptoms.

Propolis Ethanol Extract Stimulates Cytokine and Chemokine Production through NF-κB Activation in C2C12 Myoblasts.

Myoblast activation is a triggering event for muscle remodeling. We assessed the stimulatory effects of propolis, a beehive product, on myoblasts. After an 8 h treatment with 100 μg/mL of Brazilian propolis ethanol extract, expression of various chemokines, including CCL-2 and CCL-5, and cytokines, such as IL-6, increased. This propolis-induced cytokine production appears to depend on NF-κB activation, because the IKK inhibitor BMS-345541 repressed mRNA levels of CCL-2 by ~66%, CCL-5 by ~81%, and IL-6 by ~69% after propolis treatment. Supernatant from propolis-conditioned C2C12 cells upregulated RAW264 macrophage migration. The supernatant also stimulated RAW264 cells to produce angiogenic factors, including VEGF-A and MMP-12. Brazilian green propolis therefore causes myoblasts to secrete cytokines and chemokines, which might contribute to tissue remodeling of skeletal muscle.

Inhibitory Effects of Brazilian Propolis on Lipid Accumulation in 3T3-L1 Cells

We here investigated the inhibitory effect of Brazilian propolis on intracellular lipid accumulation. Seven ethanolic extracts of Brazilian propolis were tested for their inhibitory effect on intracellular lipid accumulation in 3T3-L1 cells. Among the seven Brazilian propolis, the extracts AF-08 and AF-18 had inhibitory effects on intracellular lipid accumulation. In particular, the Brazilian propolis AF-08 exhibited a potential inhibitory effect on intracellular lipid accumulation.Thus, these results suggest that the Brazilian propolis AF-08 contributes to the prevention of metabolic syndrome. In addition, five known triterpenoids were isolated from the Brazilian propolis AF-08: betulonic acid, moronic acid, 3-oxo oleanolic acid, masticadienonic acid, and anwnweizonic acid. Four of the triterpenoids exhibited an inhibitory effect on intracellular lipid accumulation. Thus, these triterpenoids may be important as antiobesity agents that inhibit the metabolic syndrome.

Protective Effect of Repeatedly Preadministered Brazilian Propolis Ethanol Extract against Stress-Induced Gastric Mucosal Lesions in Rats

The present study was conducted to clarify the protective effect of Brazilian propolis ethanol extract (BPEE) against stress-induced gastric mucosal lesions in rats. The protective effect of BPEE against gastric mucosal lesions in male Wistar rats exposed to water-immersion restraint stress (WIRS) for 6 h was compared between its repeated preadministration (50 mg/kg/day, 7 days) and its single preadministration (50 mg/kg). The repeated BPEE preadministration attenuated WIRS-induced gastric mucosal lesions and gastric mucosal oxidative stress more largely than the single BPEE preadministration. In addition, the repeated BPEE preadministration attenuated neutrophil infiltration in the gastric mucosa of rats exposed to WIRS. The protective effect of the repeated preadministration of BPEE against WIRS-induced gastric mucosal lesions was similar to that of a single preadministration of vitamin E (250 mg/kg) in terms of the extent and manner of protection. From these findings, it is concluded that BPEE preadministered in a repeated manner protects against gastric mucosal lesions in rats exposed to WIRS more effectively than BPEE preadministered in a single manner possibly through its antioxidant and anti-inflammatory actions.

Beneficial effects of Brazilian propolis on type 2 diabetes in ob/ob mice

The anti-diabetic effects of Brazilian propolis were examined using ob/ob mice. Although repeated injection of an ethanol extract of Brazilian propolis (100 mg/kg, ip, twice a week for 12 weeks) did not affect body weight gain and food intake of ob/ob mice, blood glucose and plasma cholesterol levels were significantly attenuated. Moreover, the propolis extract partially restored glucose tolerance and insulin resistance, indicating anti-diabetic properties of the extract. The propolis-treated mice exhibited lower weight gain in mesenteric adipose tissue, while weight gains in inguinal and epididymal adipose tissues were not modulated. Flow cytometric and microscopic analyses suggested that the extract promoted accumulation of eosinophils into mesenteric and epididymal adipose tissues. Alternatively, the ratio of M1-like macrophages to M2-like macrophages in mesenteric adipose tissue was reduced by the propolis injection, coincident with the decrement of the number of interleukin-12A+ cells. Levels of M1 macrophage markers, such as Itgax and Il12b transcripts, were decreased in the vascular stromal fraction of mesenteric adipose tissue, whereas those of pan-macrophage markers Emr1 and Cd68 were not influenced. Microarray and subsequent gene ontology term analyses suggested that propolis attenuated immune activation in mesenteric adipose tissues. Taken together, this indicates that Brazilian propolis improves diabetes in ob/ob mice, presumably through modification of immune cells in mesenteric adipose tissues.

Propolis extract promotes translocation of glucose transporter 4 and glucose uptake through both PI3K‐ and AMPK‐dependent pathways in skeletal muscle

It is well known that propolis has the ability to prevent hyperglycemia. However, the underlying mechanism is not yet fully understood. We therefore investigated whether a Brazilian propolis ethanol extract affects glucose uptake and translocation of insulin-sensitive glucose transporter (GLUT) 4 in skeletal muscle cells. In L6 myotubes, the extract at 1 μg/mL significantly promoted GLUT4 translocation and glucose uptake activity. Regarding the mechanism of GLUT4 translocation, propolis extract induced both PI3K and AMPK phosphorylation in a dose-dependent manner in L6 myotubes. However, we could not define which pathway was preferentially associated with GLUT4 translocation, because both PI3K and AMPK inhibitors revealed off-target effects to each other. The main polyphenols found in the propolis extract, artepillin C, coumaric acid, and kaempferide, promoted GLUT4 translocation in L6 myotubes. Additionally, these compounds activated both PI3K- and AMPK-dependent dual-signaling pathways. However, only kaempferide increased glucose uptake activity under our experimental conditions. Single oral administrations of propolis extract, at 250 mg/kg body weight, lowered postprandial blood glucose levels in ICR mice. The extract promoted GLUT4 translocation in skeletal muscle of rats and mice, but did not inhibit α-glucosidase activity in the small intestine under our experimental conditions. It was confirmed that propolis extract promoted phosphorylation of both PI3K and AMPK in rat skeletal muscle. In conclusion, we show that Brazilian propolis has the potential to prevent hyperglycemia through the promotion of GLUT4 translocation in skeletal muscle and that kaempferide is one of the candidates for active compound in propolis.

Artepillin C Suppresses Angiogenesis by Inhibiting Tube-Formation and Inducing Apoptosis of Endothelial Cells

Artepillin C (ARC) is a major active ingredient of Brazilian propolis which is a resinous mixed compound collected by honeybeesfrom various plant sources. Previously, we reported that the ethanol extract of Brazilian propolis possesses antiangiogenic activity both in vitro and in vivo. However, the mechanism of angiogenesis inhibitionby purified ARC has not been well-clarified. In this study, we investigated the effects of ARC on tube-forming human umbilical vein endothelial cells (HUVECs). We found that inhibition of tube formation by ARC was accompanied by partial fragmentation of endothelial cells, indicating that it induced cell death. Western blotting revealed that ARC induced the reduction of vascular endothelial cadherinand platelet endothelial cell adhesion molecule-1. ARC also induced chromatin condensation and cell nuclear fragmentation, morphological markers of apoptosis, in tube-forming HUVECs. Furthermore, ARC suppressed phosphorylation of extracellular signal-regulated kinase 1/2, but upregulated phosphorylation of p38. It was also shown that ARC induced apoptosis via activation of proapototic signaling, activation of caspase-3 and cleavage of poly ADP-ribose polymerase and lamin A/C. In conclusion, ARC exerts its antiangiogenic effects through induction of endothelial apoptosis.

Protective Effect of Brazilian Propolis against Liver Damage with Cholestasis in Rats Treated withα-Naphthylisothiocyanate

We examined the protective effect of Brazilian propolis against liver damage with cholestasis in rats treated with α-naphthylisothiocyanate (ANIT) in comparison with that of vitamin E (VE). Rats orally received Brazilian propolis ethanol extract (BPEE) (25, 50, or 100 mg/kg), VE (250 mg/kg) or vehicle at 12 h after intraperitoneal injection of ANIT (75 mg/kg) and were killed 24 h after the injection. Vehicle-treated rats showed liver cell damage and cholestasis, judging from the levels of serum marker enzymes and components. The vehicle group had increased serum total cholesterol, triglyceride, phospholipid, and lipid peroxide levels, increased hepatic lipid peroxide, reduced glutathione, and ascorbic acid levels and myeloperoxidase activity, and decreased hepatic superoxide dismutase activity. BPEE (50 mg/kg) administered to ANIT-treated rats prevented liver cell damage and cholestasis and attenuated these serum and hepatic biochemical changes except hepatic ascorbic acid, although administered BPEE (25 or 100 mg/kg) was less effective. VE administered to ANIT-treated rats prevented liver cell damage, but not cholestasis, and attenuated increased serum lipid peroxide level, increased hepatic lipid peroxide level and myeloperoxidase activity, and decreased hepatic superoxide dismutase activity. These results indicate that BPEE protects against ANIT-induced liver damage with cholestasis in rats more effectively than VE.

Propolis: Properties, Application, and Its Potential

Propolis: Properties, Application, and Its Potential

Inhibitory Effects of Brazilian Propolis on Tumor Promotion in Two-Stage Mouse Skin Carcinogenesis

Propolis is produced by honeybees and has many biological properties, including immunomodulatory, anti-inflammatory, anti-oxidant, anti-bacterial, anti-viral and anti-cancer actions. Five ethanol extracts of Brazilian propolis were tested for inhibition of 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation in mice. The ethanol extract of Brazilian propolis AF-08 markedly inhibited TPA-induced inflammatory ear edema in mice. This extract suppressed tumor promotion by TPA following initiation with 7,12-dimethylbenz[a]anthracene (DMBA) in mouse skin. Moronic acid, a major component of Brazilian propolis AF-08, markedly inhibited two-stage carcinogenesis by DMBA plus TPA. These results suggest that Brazilian propolis AF-08 contributes to the prevention of cancer.

Comparison of Effects of the Ethanolic Extracts of Brazilian Propolis on Human Leukemic Cells As Assessed with the MTT Assay

Propolis is a resinous product collected by honey bees. It was also reported that propolis has a wide variety of biological actions, including antimicrobial activity and antioxidant, anti-inflammatory, and suppressive effects of dioxin toxicity activities. The aim of this study was to compare the in vitro cytotoxic activities of green propolis (G12) and red propolis (G13) in human leukemia cells. These cells were incubated with different concentrations of propolis and 48 hours after the IC50 was calculated for each cell. The results showed that the red propolis has cytotoxic effect in vitro higher than green propolis. Red propolis was showed to be cytostatic in K562 cells and caused the same amount of apoptosis as its control Gleevec. In conclusion, these results showed that red propolis is more cytotoxic than the green propolis in a variety of human cell lines of leukemia. Red propolis may contain drugs capable of inhibiting cancer cell growth. Therefore, further isolation of respective chemical ingredients from the red propolis (G13) for identification of the activities is necessary.

Brazilian Propolis Suppresses Angiogenesis by Inducing Apoptosis in Tube‐Forming Endothelial Cells through Inactivation of Survival Signal ERK1/2

We recently reported that propolis suppresses tumor-induced angiogenesis through tube formation inhibition and apoptosis induction in endothelial cells. However, molecular mechanisms underlying such angiogenesis suppression by propolis have not been fully elucidated. The aim of this study was to investigate the effects of ethanol extract of Brazilian propolis (EEBP) on two major survival signals, extracellular signal-regulated kinase 1/2 (ERK1/2) and Akt, and to elucidate whether changes in these signals were actually involved in antiangiogenic effects of the propolis. Detection by western blotting revealed that EEBP suppressed phosphorylation of ERK1/2, but not that of Akt. Pharmacological inhibition by U0126 demonstrated that ERK1/2 inactivation alone was enough to inhibit tube formation and induce apoptosis. It was also shown that EEBP and U0126 similarly induced activation of caspase-3 and cleavage of poly ADP-ribose polymerase (PARP) and lamin A/C, all of which are molecular markers of apoptosis. These results indicate that inhibition of survival signal ERK1/2, and subsequent induction of apoptosis, is a critical mechanism of angiogenesis suppression by EEBP.

Abstract 765: Drug discovery for targeting the tumor microenvironment from propolis

Abstract The tumor microenvironment, characterized by regions of hypoxia, low nutrition, and acidosis due to incomplete blood vessel networks, has been recognized as a major factor that influences not only the response to conventional anti-cancer therapies but also malignant progression and metastasis. However, exploiting such a cumbersome tumor microenvironment for cancer treatment could provide tumor-specific therapeutic approaches. In particular, hypoxia is now considered a fundamentally important characteristic of the tumor microenvironment in which HIF-1 mediated gene regulation being essential for angiogenesis and tumor development. This has led to the current extensive interest in the drug discovery for tumor microenvironment including tumor specific potential molecular targets for cancer therapeutics. Propolis has pleiotropic effects and contains a variety of bioactive polyphenolic compounds. Some flavonoids of them have been reported to inhibit HIF-1α. We therefore evaluated the effects of composition of Brazilian propolis on cellular responses to hypoxia or nutrition deprivation stress for the development of novel anti-cancer drug targeting the tumor microenvironment. Materials and Method: The ethanol extract of Brazilian propolis was subjected to a series of chromatographic separations to examine HIF-1 inhibition and cytotoxicity under hypoxic and/or glucose deprived condition. The HIF-1α trans-activation effect was evaluated by HIF-1 reporter assay using stable cell lines expressing p2.1. The effect on unfolded protein response (UPR) activation in tumor cell induced by glucose deprivation or 2-deoxyglucose was also examined by GRP78 promoter reporter assay. We performed Immunoblot analysis of the HIF-1α and UPR target genes. The effects on HIF-1α dependent gene expression were investigated by RT-PCR using HCT116 cells. Result: The propolis whole extract increased HRE-dependent luciferase activity in a dose-dependent manner. From a series of chromatographic separations, we obtained fractions 3, 5 and 8 that inhibited HIF-1-dependent induction of luciferase, while fractions 6 and 10-13 increased the luciferase activity. Cinnamic acid derivative, GPU-215 was isolated from Fr.6 reduced HIF1 transactivation activity, expression of HIF-1α protein and GLUT1 mRNA under hypoxia, while flavonoid, GPU-216 from Fr.8 increased HIF-1α transactivation activity and UPR activation. Conclusion: We discovered promising compounds from Brazilian propolis for novel anti-cancer drug targeting tumor microenvironment. GPU-215 has inhibitory effect on HIF-1α expression and function, while GPU-216 increased HIF-1α transactivation and UPR activation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 765.

Anti-Influenza Virus Activity of Propolis in Vitro and its Efficacy against Influenza Infection in Mice

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. 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. 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. 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.