Table 6.
In vitro antioxidant properties of bee products.
| Bee Product | Bees Species 1 | Cell culture/Substrate | Antioxidant Activity | Measurement | References |
|---|---|---|---|---|---|
| Honey | |||||
| Monofloral honeys (Italy) | A. mellifera | Bovine brain microsomes | Peroxyl-radical scavenging capacity | Time-course of TBA-RS formation during microsomal oxidation | [74] |
| Commercial multifloral honey (Italy) | A. mellifera | Human endothelial cell line (EA.hy926) | Cell membrane oxidation, intracellular oxidative damage, cell viability using MTT [3-(4,5-dimethyl-2-thiazolyl) -2,5-diphenyl -2H- tetrazolium bromide] assay and GSH analysis | Cytoprotective activity by fluorimetric determination, cell viability (the absorbance is proportional to the number of living cells) and microscopic evaluation | [16] |
| Buckwheat and Manuka honeys | n.s. | HepG2 cell lines, Cell Bank of Institute of the Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai, China | Cellular antioxidant activity (CAA) and cytotoxicity assay | Peroxyl radical-induced oxidation of DCFH to DCF by fluorimetric determination and inhibition of oxidation by honey extracts (microscopic evaluation) | [17] |
| Malaysian kelulut honey | Trigona spp. | Lymphoblastoid cell line (LCL), AGRE, Los Angeles, CA, USA | Ferric-reducing antioxidant potential assay, total phenolic, and flavonoid content by UV spectrophotometry. Cell viability using MTS assay | Cell viability (%) reading the absorbance at 490 nm and positively affected by antioxidant properties | [18] |
| Monofloral honeys (China) | A. dorsata | HepG2 cell lines, Stem Cell Bank of Chinese Academy of Sciences | Cellular antioxidant activity (CAA) assay | Effective reduction of intracellular oxidative state reacting with peroxyl radicals or ROS/RNS. Fluorimetric determination | [19] |
| Beeswax | |||||
| Beeswax recycling by-product (MUD1) | A. mellifera | HepG2 cells, Biological Research Laboratory of Sevilla University, Spain | ROS concentration using CellROX® Orange Reagent applied according to manufacturer’s instructions. Cells were analyzed with the Tali® Image-Based cytometer | Intracellular ROS: percentage of cells with increased ROS levels related to the control | [20] |
| Two beeswax recycling by-products (MUD1 and MUD2) | A. mellifera | Adult skin HDF, GIBCO® Invitrogen cell, Waltham, MA, USA | ROS concentration using CellROX® Orange Reagent applied according to manufacturer’s instructions | Intracellular ROS: percentage of cells with increased ROS levels related to the control | [161] |
| Pollen | |||||
| Bee pollen (China) | n.s. | Blood from male Kunming mice | Superoxide dismutase (SOD) assay, lipid peroxidation index assay and total antioxidant capacity (T-AOC) assay | Spectrophotometric measurement of SOD content (U/mL), MDA content (nmol/mL) and inhibition rate (%) | [30] |
| Bee pollen from Jara pringosa (Sistus ladanifer) and Jara blanca (Cistus albidus) (Spain) | A. mellifera | Retinal ganglion cells (RGC-5, a rat ganglion cell-line transformed using E1A virus) | Antioxidant-capacity assay-measured the radicals induced in RGC-5 by the application of ROS (H2O2, O2•−, and HO) | Intracellular ROS: time-kinetic and concentration-response data for bee pollen towards production of various ROS in term of fluorescence intensity | [22] |
| Commercial pollens of different floral sources and geographical origins | n.s. | Livers obtained from pigs and homogenized | Inhibition of lipid peroxidation using thiobarbituric acid reactive substances (TBARS) | Spectrophotometric determination of inhibition ratio (%) and EC50 calculated (0.35–3.70 TBARS mg/mg extract) | [118] |
| Bee bread | |||||
| Beebread (Poland) | n.s. | Human glioblastoma cell line U87MG (HTB-14), ATCC, Rockville, MD, USA | Cytotoxicity evaluated by MTT assay. Total antioxidative ability related to phenolic and non-phenolic compounds after 24 h | Viability of U87MG (% of the control) after incubation with beebread, measuring the absorbance at 570 nm | [21] |
| Propolis | |||||
| Propolis | n.s. | Human erythrocytes from peripheral blood | Estimation of the inhibitory efficiency of propolis extracts on H2O2-induced lipid peroxidation using thiobarbituric acid (TBA) assay and protective effect of propolis extracts on H2O2-induced oxidative hemolysis | Measured the absorbance of the supernatant at 532 nm and calculated the hemolysis percentage | [47] |
| Propolis (Brazil) | M. orbignyi | Human erythrocytes from peripheral blood | Oxidative hemolysis inhibition assay, inhibitory efficiency against lipid peroxidation, cytotoxic activity and cell death profile (analysis performed using propidum iodide and annexin V-FITC dual staining) | Hemolysis (%), MDA (nmol/mL) and cell viability (%), respectively, spectrophotometrically determined and flow cytometric evaluation of death profile | [32] |
| Propolis (Portugal) | n.s. | Eukaryote unicellular model organism S. cerevisiae and human reconstituted skin tissue model (EpiDermTM EPI-200) | Evaluation of propolis protective effects against H2O2-induced oxidative stress and its influence on ROS intracellular levels in S. cerevisiae cells. UVB-induced overexpression of matrix metalloproteinases (MMPs), quantitative real-time PCR and immunohistochemistry (IHC) in skin tissue model | Viability and intracellular oxidation of S. cerevisiae cells analyzed for fluorescence by flow cytometry. Evaluation of the UVB-induced photoaging by immunohistochemistry and quantification of mRNA levels of MMPs | [142] |
| Propolis (Greece) | n.s. | Human immortalized keratinocyte (HaCaT) cell line, ATCC, Rockville, MD, USA | Determination of antioxidant capacity in cell lysates and assessment of protein oxidation by measuring the protein carbonyl colorimetric assay | DNA damage (AU) using fluorescence microscope, total antioxidant content and protein carbonyl content, spectrophotometrically determined | [23] |
| Propolis (Thailand) | n.s. | A549 human lung epithelial cells and HeLa cervical cancer cells | Determination of antioxidant activity by DPPH method and cytotoxicity by MTT assay | Extraction-method dependent antioxidant and flavonoid compounds. Cell shrinkage and floating in medium. Percentage of viability compared to the cell control | [24] |
| Propolis (Turkey) | n.s. | Human foreskin fibroblast cells (CRL-2522), ATCC, Manassas, VA, USA | Spectrofluorometric analysis of intracellular oxidative stress with CM-H2DCFDA | ROS levels measured by spectrofluorometric method | [25] |
| Brazilian green propolis from Baccharis dracunculifolia (Minas Gerais State, Brazil) | A. mellifera | Retinal ganglion cells (RGC-5, a rat ganglion cell-line transformed using E1A virus) | Antioxidant-capacity assay measured the radicals induced in RGC-5 by the application of ROS (H2O2, O2·-, and HO) | Intracellular ROS: time-kinetic and concentration-response data for propolis towards production of various ROS in terms of fluorescence intensity | [22] |
| Red propolis (Brazil) | n.s. | Human tumor cell lines HL-60 (leukemia), PC3 (prostate carcinoma), SNB19 (glioblastoma), and HCT-116 (colon carcinoma), National Cancer Institute, USA | High in vitro antioxidant activity related to total phenolic and flavonoid compound content. MTT assay to determine the cytotoxic (antitumor) potential of the extracts | Growth inhibition of tumor cell lines (%), using spectrophotometer | [26] |
| Propolis (Cameroon) | n.s. | Diluted human whole blood, mouse macrophage cell line J774.2, European Collection of Cell Cultures (UK) and NIH-3 T3 fibroblast cells, ATCC, Manassas, USA | Oxidative burst assay (luminol-enhanced chemiluminescence assay), nitric oxide assay and MTT cytotoxicity assay | ROS inhibition (EC50 µg/mL), NO inhibition (EC50 µg/mL) and cytotoxicity (EC50 µg/mL), respectively, using spectrophotometer | [33] |
| Propolis (Morocco) | n.s. | Human monocytic cell line THP-1 (ATCC 202-TIB), human colorectal carcinoma cell line HCT-116 (ATCC® CCL-247™) and breast cancer cell line MCF-7 (ATCC®HTB-22™) | High antioxidant content and activity by scavenging free radicals with IC50 (DPPH = 0.02, ABTS = 0.04, and FRAP = 0.04 mg/ml). MTT assay for cytotoxic and cytostatic activity and cell viability determination | Total phenols, flavone, and flavonol and antioxidant activity affect cell viability defined as the ratio (%) of absorbance of treated cells to untreated cells (control) | [27] |
| Propolis (Poland) | n.s. | Fresh human erythrocyte concentrates (65%), Blood bank in Poznan, Poland | High antioxidant potential related to DPPH free-radical scavenging activity and reducing power; significant protection of human red blood cells from oxidative damage. Hemolysis assays | Hemolysis (%) estimated by measuring absorbance of the supernatant; microscope studies of erythrocyte shape transformation (Bessis classification) and inhibition of free-radical-induced hemolysis | [34] |
| Royal jelly | |||||
| Enzyme-treated royal jelly (Jiangshan, China) | A. mellifera | Peritoneal macrophages, BALB/c mice | Cell viability MTT assay and ROS, SOD and GSH quantification according to the manufacturer’s kit instructions | Intracellular ROS and NO production; activity of the enzyme SOD and concentration of the antioxidant GSH (spectrophotometric quantification) | [180] |
| Fresh royal jelly from Yangtze Valley, People’s Republic of China | A. mellifera | Retinal ganglion cells (RGC-5, a rat ganglion cell-line transformed using E1A virus) | Antioxidant-capacity assay measured the radicals induced in RGC-5 by the application of ROS (H2O2, O2·-, and HO) | Intracellular ROS: time-kinetic and concentration-response data for royal jelly towards production of various ROS in terms of fluorescence intensity | [22] |
| Fresh royal jelly (Korea) and recombinant AcMRJP2 protein | A. cerana | Murine fibroblast cell line NIH 3 T3 | Antioxidant activity and shielding of the cell against oxidative stress and DNA protection against ROS. Cell viability measured by MTT assay, apoptosis assay and DNA protection assay | Antioxidant activity determines increased cell viability (%), reduced caspase-3 activity and apoptosis in the cells using laser-scanning confocal microscopy. DNA nicking assay in a metal-catalyzed oxidation system observed by agarose gel electrophoresis | [28] |
| Recombinant AmMRJPs 1–7 | A. mellifera | Murine fibroblast cell line NIH 3 T3 | Radical scavenging activity and protection against DNA oxidative damage. Cell viability measured by MTT assay, apoptosis assay and DNA protection assay | Antioxidant activity determines increased cell viability (%), reduced caspase-3 activity and apoptosis in the cells using laser-scanning confocal microscopy. DNA nicking assay in a metal-catalyzed oxidation system observed by agarose gel electrophoresis | [29] |
| Bee Venom | |||||
| Melittin (Northeast Portugal) | A. mellifera iberiensis | MCF-7, NCI-H460, HeLa and HepG2 tumour lines | Free-radical scavenging activity, reducing power, lipid peroxidation inhibition and high capacity to inhibit NO production. | Chemical characterization by LC/DAD/ESI-MS; DPPH for free-radical scavenging activity; reducing power measuring the absorbance at 690 nm | [165] |
1 A: Apis; M.: Melipona; n.s.: not specified.