Metabolomics analysis of (Geo)propolis from Brazilian stingless bees by FIA and UHPLC-HRMS (Orbitrap).

Native stingless bees (Meliponini) from Brazil make (geo)propolis which is largely used in folk medicine, specially by indigenous and quilombos communities and beekeepers´ families but are progressively being recognized for their pharmacological activities. In this study, the ethanolic extracts of (geo)propolis (EEGs) from Melipona marginata, M. quadrifasciata, M. scutellaris, and Tetragonisca angustula were analysed by Flow injection analysis (FIA) and Ultra-high performance liquid chromatography (UHPLC) in a high resolution Orbitrap mass analyser (HRMS) to investigate and compare their chemical profile. Untargeted metabolomic approach based on UHPLC-HRMS experiments, and bioinformatic tools, allowed to annotate 59 compounds from diverse classes such as: flavonoids, phenolic compounds, sugars, terpenoids, and lipids. In addition, using multivariate tools and Flow injection- high resolution mass spectrometry (FIA-HRMS), it was possible to classify samples and identify marker ions related to the bee species or genus and to the geographical origin as a proof of concept.

Commercial propolis extracts from Apis mellifera, Scaptotrigona bipunctata and Melipona quadrifasciata: quality control, antioxidant and antiglycemic potential.

Quality control of propolis plays a pivotal role in ensuring the appropriate concentrations of active compounds, limiting unwanted substances, verifying authenticity, and adhering to regulatory standards. This study aimed to assess the identity and quality standards, the individual phenolic composition (LC-ESI-MS/MS), and the antioxidant and antiglycemic potential of commercial propolis extracts (CPEs) from Apis mellifera, Scaptotrigona bipunctata, and Melipona quadrifasciata bees. CPEs met wax content and oxidation activity criteria, surpassing minimum thresholds for total phenolic content (TPC) and flavonoid content (TFC), although stingless bee CPE did not test positive for 10% lead acetate. CPEs exhibited antioxidant and potential antiglycemic activities. Epicatechin among the thirty-three identified phenolics, showed significant correlation with TPC, DPPH, ABTS, and EC50 values of α-amylase enzyme. These promising attributes underscore the potential health benefits of commercial propolis extracts from Apis mellifera and stingless bees for further medicinal and nutritional applications.

Anti-inflammatory action of geopropolis produced by stingless bees on human peripheral blood mononuclear cells.

Natural products have been used to treat inflammatory reactions and led to the discovery of new anti-inflammatory drugs. Geopropolis (GEO) is produced by stingless bees and has been used by indigenous people to improve the immune functions. Here, a possible synergism between GEO and dexamethasone (DEX) was assessed on human peripheral blood mononuclear cells (PBMC) stimulated by lipopolysaccharide (LPS). PBMC viability was evaluated by the MTT, apoptosis/necrosis by flow cytometry, cytokine and eicosanoids production by ELISA, and intracellular pathways by polymerase chain reaction. GEO and DEX alone or in combination did not affect cell viability. GEO in combination with lower concentrations of DEX inhibited cytokine production (TNF-α, IL-1ß, and IL-10). No effects were seen on eicosanoids nor in intracellular pathways. Despite not always being more efficient than the isolated treatments, GEO + DEX seemed to be promising and allow the use of DEX in lower concentrations, reducing adverse effects.

Antiviral Activity of Flavonoids from Geopropolis of the Brazilian Jandaira Bee against Zika and Dengue Viruses.

Arthropod-borne viruses within the Flaviviridae family such as Zika (ZIKV) and dengue (DENV) are responsible for major outbreaks in tropical countries, and there are no specific treatments against them. Naringenin and 7-O-methyl naringenin are flavonoids that can be extracted from geopropolis, a natural material that the Brazilian Jandaira stingless bee (Melipona subnitida Ducke) produces to protect its nest. Here, these flavonoids were tested against ZIKV and DENV using Vero cells as a cellular model to perform a cytotoxicity assay and to define the effective concentrations of TCID50 as the readout method. The results demonstrated the antiviral activity of the compounds against both viruses upon the treatment of infected cells. The tested flavonoids had antiviral activity comparable with 6-methylmercaptopurine riboside (6-MMPr), used here as a positive control. In addition, to identify the possible action mechanism of the antiviral candidates, we carried out a docking analysis followed by a molecular dynamics simulation to elucidate naringenin and 7-O-methyl naringenin binding sites to each virus. Altogether, these results demonstrate that both flavonoids have potent antiviral effects against both viruses and warrant further in vivo trials.

Antibacterial and Toxic Activity of Geopropolis Extracts from Melipona subnitida (Ducke, 1910) (Hymenoptera: Apidae) and Scaptotrigona depilis (Moure, 1942) (Hymenoptera: Apidae).

Bacteria are associated with many infections that affect humans and present antibiotic resistance mechanisms, causing problems in health organisations and increased mortality rates. Therefore, it is necessary to find new antibacterial agents that can be used in the treatment of these microorganisms. Geopropolis is a natural product from stingless bees, formed by a mixture of plant resins, salivary secretions, wax and soil particles, the chemical composition of this natural product is diverse. Thus, this study aimed to evaluate antibacterial activity, antibiotic modulation and the toxicity of geopropolis extracts from the stingless bees, Melipona subnitida (Ducke, 1910) and Scaptotrigona depilis (Moure, 1942) against standard and multi-resistant Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa bacteria. Geopropolis samples were collected in a meliponary located in Camaragibe, Pernambuco, Brazil. To determine the Minimum Inhibitory Concentration (MIC) and antibiotic modulation we performed broth microdilution tests. Mortality tests were used to verify extract toxicity in the model Drosophila melanogaster. The microbiological tests showing that the M. subnitida extracts had better inhibitory effects compared to S. depilis, presenting direct antibacterial activity against standard and multi-resistant strains. The extracts potentialized antibiotic effects, suggesting possible synergy and did not present toxicity in the model used. The information obtained in this study highlights extracts as promising antibacterial agents and is the first study to evaluate bacterial activity in these extracts, in addition to verifying their modulating effects and determining toxicity in the model used.

The chemical composition and antioxidant activity of mandaçaia (melipona quadrifasciata) geopropolis varies more due to region than month of collection.

Stingless bees are responsible for pollinating up to 90% of Brazilian flora, so their study is of commercial and ecological importance. Stingless bees mix plant resins with wax and soil, forming geopropolis. Studies of the variability in composition and activity due to species, place and season of collection are lacking. Yield, total phenolic and flavonoid content, and antioxidant activity (via DPPH) of the extracts of fifteen M. quadrifasciata (Mandaçaia) geopropolis samples collected over a year in the State of São Paulo, Brazil, as well as two samples from different regions of the state of Minas Gerais in Brazil were compared. Composition was determined by ultra-high-resolution liquid chromatography and mass spectrometry, identifying 16 compounds. The month and region of collection affected the available plant resins and influenced their yield, composition and antioxidant capacity. Depending on the place of collection, M. quadrifasciata geopropolis is a promising natural source of antioxidant phenolic compounds.

Use of Stingless Bee Propolis and Geopropolis against Cancer-A Literature Review of Preclinical Studies.

Cancer is one of the major maladies affecting humankind and remains one of the leading causes of death worldwide. The investigation of the biological activities of stingless bee products, especially propolis and geopropolis, has revealed promising therapeutic properties, especially in the research on new antineoplastic agents. This literature review of preclinical trials, involving biological assays of antitumor activity and identification of the chemical composition of propolis and geopropolis of stingless bee species, describes the cytotoxicity in tumor lineages (breast, lung, ovarian, liver, mouth, pharynx, larynx, colon, stomach, colorectal, cervix, kidney, prostate, melanoma, human glioblastoma, canine osteosarcoma, erythroleukemia, human chronic myelocytic leukemia, and human promyelocytic leukemia) of propolis and geopropolis of 33 species of stingless bees. The chemical composition of propolis and geopropolis was identified, indicating that these belong to the chemical classes of phenolic acids, flavonoids, coumarins, benzophenones, anthraquinones, alkaloids, terpenes, steroids, saponins, fatty acids, and carbohydrates and are possibly responsible for the cytotoxicity in tumor cells. Apoptosis was one of the main mechanisms of cytotoxicity of extracts and substances isolated from stingless bee products. Although the results found are encouraging, other preclinical studies and clinical trials are essential for the discovery of new anticancer agents.

Essential and Potentially Toxic Elements from Brazilian Geopropolis Produced by the Stingless Bee Melipona quadrifasciata anthidioides Using ICP OES.

Melipona quadrifasciata anthidioides is a species of stingless bee popularly known in Brazil as "mandaçaia". Products derived from bees for food and therapeutic uses, have stimulated the evaluation of the chemical composition of geopropolis. Concentrations of 24 essential and potentially toxic elements were determined in geopropolis samples, using a sequential optical emission spectrometer with inductively coupled plasma (ICP OES) after microwave-assisted acid digestion. Principal component analysis (PCA) and Hierarchical cluster analysis (HCA) were used to carry out an exploratory analysis of the samples. The following elements were quantified (in mg Kg-1): Al (320,414.40-36,911.1), As (

A comprehensive characterization of polyphenols by LC-ESI-QTOF-MS from Melipona quadrifasciata anthidioides geopropolis and their antibacterial, antioxidant and antiproliferative effects.

The geopropolis is a unique type of propolis produced by some stingless bee species. This product is known in folk medicine for its pharmacological properties, mainly antimicrobial and antioxidant, but there are few scientific studies that prove these properties. The objective of this study was to evaluate the phenolic composition and the antimicrobial, antioxidant and antiproliferative activities of Melipona quadrifasciata geopropolis. The phenolic characterization of the geopropolis ethanolic extract was evaluated by LC-ESI-QTOF-MS. The antimicrobial activity was carried out against Gram-positive (including multiresistant microorganisms), negative and yeast. The synergistic effect was evaluated in association with Sulfamethoxazole + Trimethoprim. DPPH, ABTS, FRAP, ORAC and HPLC on-line were used to evaluate the antioxidant activity. Antiproliferative activity was assessed by the sulforhodamine B assay. Flavonoids and phenolic acids were identified in the extract, which showed promising antimicrobial activity, partially synergistic effect and antioxidant activity.

Antimicrobial, anti-adherence and antibiofilm activity against Staphylococcus aureus of a 4-phenyl coumarin derivative isolated from Brazilian geopropolis.

The discovery of new drug candidates, especially from natural products, remains a promising approach to overcome the alarmingly high microbial resistance rates. A major 4-phenyl coumarin named cinnamoyloxy-mammeisin (CNM) isolated from stingless bee geopropolis showed interesting biological properties; however, its antimicrobial activity against Staphylococcus aureus has never been investigated. In order to clarify these properties, CNM isolated from geopropolis was initially tested against methicillin-susceptible and -resistant S. aureus strains. Further, the effects of CNM were assessed on the microbial adherence to human cells, biofilm formation and mature biofilm. Then, the acute toxicity of the compound was determined in Galleria mellonella. CNM showed bacteriostatic activity against methicillin-susceptible and -resistant S. aureus strains, with MIC of 11.3 µM. In addition, CNM at 5.7 µM reduced bacterial adherence to human keratinocytes from 1 to 3 h and disrupted biofilm formation by reducing cell viability and architecture, as evidenced by scanning electron microscopy. The acute toxicity assay indicated no significant harmful effects. Based on these findings, CNM can be considered a promising compound with anti-S. aureus properties and predicted low toxicity. Thus, it may be used as a drug candidate or lead compound for structure/activity optimization.

Brazilian stingless bee propolis and geopropolis: promising sources of biologically active compounds

ABSTRACT Stingless bee products such as honey, pollen, propolis, and geopropolis have been used for centuries in traditional medicine for the treatment of several illnesses. Investigation of the biological activity of stingless bee products, especially propolis and geopropolis, has revealed promising therapeutic properties. About 20% of total Neotropical stingless bees can be found in Brazil. Despite the species diversity, studies on their biological activity are scarce. The present review focuses on the antioxidant and antimicrobial activities of propolis and geopropolis from Brazilian stingless bees. In addition, the toxicity of these natural products was addressed. In order to provide new evidences for the toxic potential of propolis and geopropolis components, an in silico analysis was performed using the ADMET PredictorTM software. We observed that most of studies evaluated only crude ethanol extracts of a limited number of stingless bees species. Propolis and geopropolis displayed antioxidant capacity and antimicrobial activity. Concerning the toxic potential, the extracts of stingless bees propolis and geopropolis were considered safe. Nonetheless, in vitro and in vivo toxicological studies are still necessary.

Antileishmanial activity and chemical composition from Brazilian geopropolis produced by stingless bee Melipona fasciculata

ABSTRACT Geopropolis is produced by some stingless bee species such as Melipona fasciculata and consists of a mixture of plant resins, salivary secretions of the bee, wax, and soil. This study evaluated the antileishmanial activity in vitro, cytotoxicity and chemical composition of geopropolis produced by M. fasciculata in the savannah region of Maranhão, Brazil. The geopropolis extract was obtained through maceration using in 70% ethanol. The hydroalcoholic extract of geopropolis after liquid-liquid partition yielded the hexane, chloroformic, ethyl acetate and hydroalcoholic fractions. Antileishmanial activity was evaluated against promastigote and intracellular amastigote of Leishmania amazonensis. Cytotoxic was realized in BALB/c mice peritoneal macrophages. Chemical analysis was performed by gas chromatography-mass spectrometry and high performance liquid chromatography coupled to an ultraviolet-visible detector. The geopropolis inhibited the L. amazonensis promastigotes growth and was effective in reducing the infection of murine macrophages since the number of internalized amastigotes were smaller in cells treated with the geopropolis extract in relation to the untreated group. The ethyl acetate fraction was the most active and showed the highest index of selectivity as antileishmanial product. The geopropolis from M. fasciculata had an antileishmanial effect, mainly after the obtention of the ethyl acetate fraction that improved the activity without increasing the cytotoxicity against murine macrophages. Analysis for gas chromatography-mass spectrometer identified as main compounds the gallic and ellagic phenolic acids, either in the extract or in the active fraction. The results obtained by high performance liquid chromatography it was possible to confirm the presence and quantify the concentration gallic and ellagic acid either in the extract or in the active fraction. These results suggest that the antileishmanial activity of geopropolis is related to the presence of derivatives of these phenolic acids, mainly gallic and ellagic acids.

Geopropolis gel for the adjuvant treatment of candidiasis - formulation and in vitro release assay

ABSTRACT The geopropolis produced by the stingless bee Melipona subnitida (popularly called "jandaíra" in Brazil) is a mixture of resin, wax, and mud. This study analyzed the antifungal activity of the geopropolis extract from Candida spp., developed a gel formulation with this extract and analyzed the delivery of bioactives (kinetics release) in the formulation and their chemical profile by UHPLC-PDA-qTOF-MS/MS. Three different gels were prepared using the geopropolis extract, carbomer, propylene glycol, and water. Formulations with different amounts of propylene glycol were investigated. Physical, visual, pH, viscosity, adhesion, spreadability, leakage, and in vitro release tests were performed in the proposed formulations. Antifungal tests with the geopropolis ethanolic extract were carried out against six Candida species. The chemical profile of the geopropolis extract and compounds released from the formulations were analyzed after the release test. The formulations had a pH between 4.6 and 4.8 and viscosity between 535,600 and 920,400 cPs. The geopropolis extract presented excellent antifungal activity against the tested yeasts. The results of the release test in semipermeable cellulose membrane showed that all formulations containing 5%, 10% and 40% propylene glycol presented release of geopropolis extract. For adhesion and leakage tests, the gel formulation with 5% propylene glycol was more effective. Both geopropolis ethanolic extract and the liquid obtained in the release test showed the presence of flavonoids (flavonol/flavone, flavanone, and chalcones). Gel formulations with geopropolis extract that are rich in flavonoids are promising as an adjuvant treatment of vaginal candidiasis.

A new chemotherapeutic approach using doxorubicin simultaneously with geopropolis favoring monocyte functions.

AIMS: Chemotherapy has been widely used to treat cancer although it may affect non-target cells involved in the immune response. This work aimed at elucidating whether the chemotherapeutic agent doxorubicin in combination with geopropolis produced by Melipona fasciculata Smith could affect nontumor immune cells, evaluating their immunomodulatory effects on human monocytes. MAIN METHODS: Cell viability, expression of cell markers (HLA-DR, TLR-2, TLR-4, C80 and CD40), cytokine production (TNF-α, IL-1ß, IL-6 and IL-10), intracellular pathways (NF-κB and autophagy), the microbicidal activity of monocytes and hydrogen peroxide (H2O2) production were analyzed. KEY FINDINGS: Data showed that doxorubicin + geopropolis diminished IL-6 secretion, stimulated TNF-α and IL-10 production, TLR-4 and CD80 expression, NF-κB and autophagy pathway, as well as the bactericidal activity. SIGNIFICANCE: Our findings revealed a new chemotherapeutic approach using doxorubicin simultaneously with geopropolis without affecting human monocytes viability and exerting immunomodulatory effects, favoring cell functions. While doxorubicin altered some immunological parameters, the addition of geopropolis compensated some changes.

Characterisation of phenolic compounds by UPLC-QTOF-MS/MS of geopropolis from the stingless bee Melipona subnitida (jandaíra).

INTRODUCTION: Melipona subnitida Ducke (jandaíra) is a stingless bee native to north-eastern Brazil, which produces geopropolis, a mixture of beeswax, plant resins, pollens and earth that is used for sealing beehives. OBJECTIVE: To extend the knowledge on phenolic compounds in fractions obtained by C18-solid phase extraction (SPE) of nine geopropolis samples from Melipona subnitida collected at different times. METHODOLOGY: Chromatographic profiles of nine samples of geopropolis from jandaíra were analysed by ultra-performance liquid chromatography coupled with a diode array detector and quadrupole time-of-flight mass spectrometry (UPLC-DAD-QTOF-MS/MS) and combined with the use of data-independent acquisition (MSE) for the profiling and structural characterisation of the phenolic compounds. The isolated compound was identified by nuclear magnetic resonance of hydrogen and carbon (1 H- and 13 C-NMR). RESULTS: The present study with geopropolis of jandaíra resulted in the characterisation of 51 phenolics by UPLC-DAD-QTOF-MS/MS: four galloyl glucosides, one ellagic acid, 11 acyl-hexosides, 23 acyl-galloyl-hexosides and 12 flavonoids. The structures of two compounds (1,6-di-O-(E)-coumaroyl-2-O-galloyl-ß-d-glucopyranoside and 1-O-cinnamoyl-6-O-(E)-coumaroyl-2-O-galloyl-ß-d-glucopyranoside) were established by 1 H and the attached proton test (APT) experiments as well as high-resolution electrospray ionisation mass spectroscopy (HR-ESI-MS) analysis. CONCLUSION: The geopropolis of jandaíra showed phenolic compounds galloyl hexosides, ellagic acid, acyl-(cinnamoyl/coumaroyl)-hexosides, acyl-(cinnamoyl/coumaroyl)-galloyl-hexosides and flavonoids (aglycones and acylated-O-glycosides).

Metals in geopropolis from beehive of Melipona scutellaris in urban environments.

Geopropolis, a different type of propolis, presents a mixture of resin and exudates, containing wax, silt, and sand particles. This product has been the subject of research interest for its physicochemical properties, economic importance, and likely for environmental monitoring. The determination of toxic metals in hive products has been reported as an efficient tool for environmental monitoring. As the honey production is now common in urban environments, this study aimed to determine the concentration of toxic metals in the Melipona scutellaris geopropolis of the Metropolitan Region of Salvador, Bahia. Geopropolis and soil samples were collected from seven important beehives between June 2015 and July 2016. After EPA 3050B acid digestion procedure, metals were determined by ICP OES. As the geopropolis is partially made from soil, the values of Cr and Fe were extremely more important than the values reported in propolis, wax, and honey found in other worldwide studies. It gives different characteristics to this product depending on the location of the hive and characterizes it as an efficient integrating indicator of soil pollution. Using the enrichment factor, we determined that the soils around the meliponary are not or only slightly polluted by Cu, Cr, Ni, Pb and Zn. However, there was a shift in the particle size of geopropolis, being loamier and less sandy than the surrounding soil. In such case, a higher metal content could be expected, though no metal enrichment in the geopropolis was found, even a decrease in zinc, possibly due to the exclusion of metals by bees, was noted. Nevertheless, the results on the proportions of lithogenic metal and anthropogenic metal indicate that some metals have an external origin (about 20% for Ni and Cu). Geopropolis can be considered as a good environmental indicator even in low contaminated areas.

Antiviral activity of geopropolis extract from Scaptotrigona Aff. postica against rubella virus

The search for functional foods, which possess bioactive substances, is a new trend for the obtention of alternative and more effective treatments of many diseases with fewer side effects. Geopropolis, elaborated by stingless bees, is a mixture of plant resin sources, wax and soil. In the geopropolis from Scaptotrigona affinis postica (Latreille, 1807), (Hymenoptera, Apidae, Meliponini) was not observed the presence of soil. In a previous study, the extract of geopropolis provided by the beekeeper, from S. postica of Barra do Corda, Maranhão State, exhibited potent antiviral activity against herpes simplex virus. In this study, the propolis extract was prepared experimentally and characterized by RP-HPLC-DAD-ESI-MS/MS. The objective of this study was to evaluate the antiviral activity of an experimentally prepared geopropolis extract from S. postica against Rubella Virus infected Statens Serum Institute Rabbit Cornea (SIRC) cells. Rubella virus infection of susceptible women during the first trimester of pregnancy, often results in a combination of birth defects in newborns. There is not an effective treatment for rubella virus infection. Different protocols were carried out to evaluate, the antiviral effect of geopropolis extract on the viral replication of infectious RV. Cell viability and cell proliferation assays indicated that this geopropolis was not toxic to cultured SIRC cells. In the viral binding assay, antiviral assay, real-time PCR, and transmission electron microscopy, was observed that different concentrations of geopropolis (17, 34 and 68 µg/mL) was able to inhibit the binding of virions to the cell receptor and the production of infectious RV particles in post treated and pre treated infected SIRC cells. The antiviral activity could to be attributed to the high contents of the apigenin derivatives, vicenin-2 and schaftoside. As far as we know, this is the first report about the antiviral activity of geopropolis from Scaptotrigona postica against a Togaviridae virus.

Melipona mondury produces a geopropolis with antioxidant, antibacterial and antiproliferative activities

ABSTRACT Geopropolis is a special type of propolis produced by stingless bees. Several pharmacological properties have been described for different types of geopropolis, but there have been no previous studies of the geopropolis from Melipona mondury. In this study, we investigated the antioxidant, antibacterial, and antiproliferative activities of M. mondury geopropolis, and determined its chemical profile. The antioxidant activity was determined using in vitro ABTS·+, ·DPPH, and β-carotene/linoleic acid co-oxidation methods. The antibacterial activity was determined using a microdilution method with Pseudomonas aeruginosa, Staphylococcus aureus, and methicillin-resistant S. aureus. The antiproliferative effect was determined in tumor cell lines using the Alamar Blue assay. The chemical profile was obtained using UHPLC-MS and UHPLC-MS/MS. The butanolic fraction had the highest concentration of phenolic compounds and more potent antioxidant properties in all assays. This fraction also had bacteriostatic and bactericidal effects against all bacterial strains at low concentrations, especially S. aureus. The hexane fraction had the highest antiproliferative potential, with IC50 values ranging from 24.2 to 46.6 µg/mL in HL-60 (human promyelocytic leukemia cell) and K562 (human chronic myelocytic leukemia cell), respectively. Preliminary chemical analysis indicates the presence of terpenes and gallic acid in the geopropolis. Our results indicate the therapeutic potential of geopropolis from M. mondury against inflammatory, oxidative, infectious, and neoplastic diseases.

Pythium insidiosum: inhibitory effects of propolis and geopropolis on hyphal growth

Abstract Propolis and geopropolis are resinous products of bees showing antimicrobial effects. There is no data concerning their action against Pythium insidiosum - the causative agent of pythiosis, a pyogranulomatous disease of the subcutaneous tissue that affects mostly horses, dogs and humans. Fragments of 15 isolates of P. insidiodum were incubated with propolis and geopropolis extracts and evaluated for up to seven days to detect the minimal fungicidal concentration (MFC). Propolis inhibited three isolates at 1.0 mg mL-1 after 24 h and all other isolates at 3.4 mg mL-1. Geopropolis led to more variable results, exerting predominantly a fungistatic action than a fungicidal one. Propolis was more efficient than geopropolis in inhibiting P. insidiosum since lower concentrations led to no growth after 24 h. This effect may be due to propolis chemical composition, which has more active compounds than geopropolis. Propolis seemed to be a good candidate for in vivo studies, since treatment with conventional antifungal compounds is difficult in most of the cases, requiring extensive surgical debridement.

Immunomodulatory/inflammatory effects of geopropolis produced by Melipona fasciculata Smith in combination with doxorubicin on THP-1 cells.

OBJECTIVES: Geopropolis (GEO) in combination with doxorubicin (DOX) reduced HEp-2 cells viability compared to GEO and DOX alone. A possible effect of this combination on the innate immunity could take place, and its effects were analysed on THP-1 cell - a human leukaemia monocytic cell line used as a model to study monocyte activity and macrophage activity, assessing cell viability, expression of cell markers and cytokine production. METHODS: THP-1 cells were incubated with GEO, DOX and their combination. Cell viability was assessed by MTT assay, cell markers expression by flow cytometry and cytokine production by ELISA. KEY FINDINGS: GEO + DOX did not affect cell viability. GEO alone or in combination increased TLR-4 and CD80 but not HLA-DR and TLR-2 expression. GEO stimulated TNF-α production while DOX alone or in combination did not affect it. GEO alone or in combination inhibited IL-6 production. CONCLUSIONS: GEO exerted a pro-inflammatory profile by increasing TLR-4 and CD80 expression and TNF-α production, favouring the activation of the immune/inflammatory response. GEO + DOX did not affect cell viability and presented an immunomodulatory action. Lower concentrations of DOX combined to GEO could be used in cancer patients, avoiding side effects and benefiting from the biological properties of GEO.