Review of Therapeutic Effects of Bee Venom on Diseases Such as Asthma, Parkinson's Disease

Bee Venom Phospholipase A₂: Yesterday's Enemy Becomes Today's Friend

Toxins (Basel). 2016 Feb 22;8(2)

Bee venom therapy has been used to treat immune-related diseases such as arthritis for a long time. Recently, it has revealed that group III secretory phospholipase A₂ from bee venom (bee venom group III sPLA₂) has in vitro and in vivo immunomodulatory effects.

A growing number of reports have demonstrated the therapeutic effects of bee venom group III sPLA₂. Notably, new experimental data have shown protective immune responses of bee venom group III sPLA₂ against a wide range of diseases including asthma, Parkinson's disease, and drug-induced organ inflammation. It is critical to evaluate the beneficial and adverse effects of bee venom group III sPLA₂ because this enzyme is known to be the major allergen of bee venom that can cause anaphylactic shock.

For many decades, efforts have been made to avoid its adverse effects. At high concentrations, exposure to bee venom group III sPLA₂ can result in damage to cellular membranes and necrotic cell death.

In this review, we summarized the current knowledge about the therapeutic effects of bee venom group III sPLA₂ on several immunological diseases and described the detailed mechanisms of bee venom group III sPLA₂ in regulating various immune responses and physiopathological changes.

Bee Venom Component Inhibits Tumor Cell Growth

Melittin suppresses cathepsin S-induced invasion and angiogenesis via blocking of the VEGF-A/VEGFR-2/MEK1/ERK1/2 pathway in human hepatocellular carcinoma

Oncol Lett. 2016 Jan;11(1):610-618

Melittin, a significant constituent of Apis mellifera (honeybee) venom, is a water-soluble toxic peptide that has traditionally been used as an antitumor agent. However, the underlying mechanisms by which it inhibits tumor cell growth and angiogenesis remain to be elucidated.

In the present study, screening for increased cathepsin S (Cat S) expression levels was performed in MHCC97-H cells and various other hepatocellular carcinoma cell lines by reverse transcription-polymerase chain reaction and western blot analysis. A pcDNA3.1-small hairpin RNA (shRNA)-Cat S vector was stably transfected into MHCC97-H cells (shRNA/MHCC97-H) in order to knockdown the expression of Cat S. The effects resulting from the inhibition of Cat S-induced proliferation, invasion and angiogenesis by melittin were examined using cell proliferation, cell viability, flat plate colony formation, migration, wound healing, Transwell migration and ELISA assays. In order to substantiate the evidence for melittin-mediated inhibition of Cat S-induced angiogenesis, Cat S RNA was transfected into primary human umbilical vein endothelial cells (Cat S-HUVECs) to induce overexpression of the Cat S gene.

The effects of melittin on HUVECs were examined using Transwell migration and tube formation assays. The findings demonstrated that melittin was able to significantly suppress MHCC97-H cell (Mock/MHCC97-H) proliferation, invasion and angiogenesis, as well as capillary tube formation of Cat S-HUVECs, in a dose-dependent manner. However, proliferation, invasion and angiogenesis in shRNA/MHCC97-H and in native HUVECs (Mock-HUVECs) were unaffected. In addition, melittin specifically decreased the expression of phosphorylated (activated) Cat S, and components of the vascular endothelial growth factor (VEGF)-A/VEGF receptor 2 (VEGFR-2)/mitogen-activated protein kinase kinase 1 (MEK1)/extracellular signal-regulated kinase (ERK)1/2 signaling pathway in Mock/MHCC97-H cells.

In conclusion, the inhibition of tumor cell growth and anti-angiogenic activity exerted by melittin may be associated with anti-Cat S actions, via the inhibition of VEGF-A/VEGFR-2/MEK1/ERK1/2 signaling.

Propolis May Help Treat Insulin-Resistance Related Diseases

Caffeic Acid Phenethyl Ester Regulates PPAR's Levels in Stem Cells-Derived Adipocytes

PPAR Res. 2016;2016:7359521

Hypertrophic obesity inhibits activation of peroxisome proliferators-activated receptor gamma (PPARγ), considered the key mediator of the fully differentiated and insulin sensitive adipocyte phenotype. We examined the effects of Caffeic Acid Phenethyl Ester (Cape), isolated from propolis, a honeybee hive product, on Adipose Stem Cells (ASCs) differentiation to the adipocyte lineage. Finally we tested the effects of Cape on insulin-resistant adipocytes.

Quantification of Oil Red O-stained cells showed that lipid droplets decreased following Cape treatment as well as radical oxygen species formation. Additionally, exposure of ASC to high glucose levels decreased adiponectin and increased proinflammatory cytokines mRNA levels, which were reversed by Cape-mediated increase of insulin sensitivity. Cape treatment resulted in decreased triglycerides synthesis and increased beta-oxidation. Exposure of ASCs to Lipopolysaccharide (LPS) induced a reduction of PPARγ, an increase of IL-6 levels associated with a well-known stimulation of lipolysis; Cape partially attenuated the LPS-mediated effects.

These observations reveal the main role of PPARγ in the adipocyte function and during ASC differentiation. As there is now substantial interest in functional food and nutraceutical products, the observed therapeutic value of Cape in insulin-resistance related diseases should be taken into consideration.

Gelam (Melaleuca ) Honey Promotes Corneal Wound Healing

Gelam honey potentiates ex vivo corneal keratocytes proliferation with desirable phenotype expression

BMC Complement Altern Med. 2016 Feb 24;16(1):76

BACKGROUND:

This study aimed to evaluate the effects of Gelam honey on corneal keratocytes proliferative capacity and phenotypic characterization via MTT assay, gene expression and immunocytochemistry.

METHODS:

Corneal keratocytes from New Zealand white rabbits were cultured in basal medium (BM) and serum enriched medium (BMS). Serial dilutions of Gelam honey (GH) were added to both media and cells were cultured until passage 1. MTT assay was performed on corneal keratocytes in both media to ascertain the optimal dose of GH that produced maximum proliferation.

RESULTS:

Gelam honey at the concentration of 0.0015 % in both media showed the highest proliferative capacity with no morphological changes compared to their respective controls. The gene expression of aldehyde dehydrogenase (ALDH), a marker for quiescent keratocytes and vimentin, a marker for fibroblast, were higher in the GH enriched groups. The alpha smooth muscle actin (α-SMA) expression, marker for myofibroblast, was lower in GH treated groups compared to the controls. Immunocytochemistry results were in accordance to the gene expression analyses.

CONCLUSION:

Gelam honey at a concentration of 0.0015 % promotes ex vivo corneal keratocytes proliferation while retaining desirable phenotype expression. The results serve as a basis for the development of Gelam honey as a potential natural product in promoting corneal wound healing.

Brazilian Firm Introduces Propolis Gel to Treat Mouth Infections in Cancer Patients

Company introduces propolis dental gel at Gulfood

Brazil-Arab News Agency, 2/23/2016

Dubai – A first-timer at Gulfood, the food industry expo running until the 25th in Dubai, Pharma Nectar, a company from Brazil’s Minas Gerais state, is offering a rather unique product for a trade show of this type: a propolis-based dental gel. The result of ten years of research and testing, the gel fights mouth infections in cancer patients. The company is exhibiting at the pavilion organized by the Brazilian Export and Investment Promotion Agency (Apex-Brasil).

“Worldwide, 6% of patients with head or shoulder cancers die from opportunistic infections of the buccal cavity,” explained CEO José Alexandre Silva de Abreu. The gel was developed in partnership with the Federal University of Minas Gerais (UFMG) and the University of Campinas (Unicamp).

According to him, radiotherapy, a treatment for fighting cancerous cells, causes burns on the salivary glands, rendering patients unable to produce saliva. “The saliva controls mouth microbes, and without it, bacterial counts soar,” he pointed out.

The product was launched in 2013. The following year, it won Phama Nectar the Apex-Brasil Prize for Export Excellence in the micro/small business category. According to Abreu, the gel’s development cost roughly BRL 4 million (USD 993,488 at current exchange rates). It is a natural, preservative-free product and can be ingested.

“Propolis has antibacterial, healing and anti-inflammatory properties, and it boosts patient immunity,” said director Sheila Abreu. The product is currently sold in the United States, Greece, France, Japan and Korea. In the Us, a 90 gram tube sells for USD 40 on average.

Review of Therapeutic Effects of Bee Venom on Asthma, Parkinson’s Disease

Bee Venom Phospholipase A2: Yesterday’s Enemy Becomes Today’s Friend

Toxins 2016, 8(2), 48

Bee venom therapy has been used to treat immune-related diseases such as arthritis for a long time. Recently, it has revealed that group III secretory phospholipase A2 from bee venom (bee venom group III sPLA2) has in vitro and in vivo immunomodulatory effects.

A growing number of reports have demonstrated the therapeutic effects of bee venom group III sPLA2. Notably, new experimental data have shown protective immune responses of bee venom group III sPLA2 against a wide range of diseases including asthma, Parkinson’s disease, and drug-induced organ inflammation.

It is critical to evaluate the beneficial and adverse effects of bee venom group III sPLA2 because this enzyme is known to be the major allergen of bee venom that can cause anaphylactic shock. For many decades, efforts have been made to avoid its adverse effects. At high concentrations, exposure to bee venom group III sPLA2 can result in damage to cellular membranes and necrotic cell death.

In this review, we summarized the current knowledge about the therapeutic effects of bee venom group III sPLA2 on several immunological diseases and described the detailed mechanisms of bee venom group III sPLA2 in regulating various immune responses and physiopathological changes.

Honey, Defensin-1 Effective Against Biofilm

Antibiofilm efficacy of honey and bee-derived defensin-1 on multi-species wound biofilm

Published Ahead of Print: 08 February, 2016 Journal of Medical Microbiology doi:

Many clinically relevant biofilms are polymicrobial. Examining the effect of antimicrobials in a multi-species biofilm consortium is of great clinical importance. The goal of this study was to investigate the effect of different honey types against bacterial wound pathogens grown in multi-species biofilm and to test the antibiofilm activity of honey defensin-1 in its recombinant form.

Modified Lubbock chronic wound biofilm formed by four bacterial species (Staphylococcus aureus, Streptococcus agalactiae, Pseudomonas aeruginosa, Enterococcus faecalis) was used for evaluation of honey and recombinant bee-derived defensin-1 antibiofilm efficacy.

Defensin-1 was prepared by heterologous expression in Escherichia coli. We showed that different types of honey (manuka and honeydew) were able to significantly reduced cell viability of wound pathogens (S. aureus, S. agalactiae and P. aeruginosa) in polymicrobial biofilm. None of the tested honeys showed the ability to eradicate E. faecalis in biofilm. In addition, recombinant defensin-1 successfully reduced the viability of S. aureus and P. aeruginosa cells within established polymicrobial biofilm after 24 and 48 hours of treatment. Interestingly, recombinant defensin-1 did not affect the viability of S. agalactiae cells within the biofilm whereas both natural honeys significantly reduced the viable bacteria. Although E. faecalis was highly resistant to defensin-1, defensin-1 significantly affected biofilm formation of E. faecalis and S. agalactiae after 24 hours of treatment, most likely through the inhibiting its extracellular polymeric substances production.

In conclusion, our study reveals that honey and defensin-1 are effective against established multi-species biofilm; however, E. faecalis grown in multi-species biofilm was resistant to both antimicrobials.

Antibacterial Activity of Digested Manuka Honey Studied

Antistaphylococcal activity and metabolite profiling of manuka honey (Lectospermum scoparium L.) after in vitro simulated digestion

Food Funct., 2016, Accepted Manuscript
First published online 18 Feb 2016

The antistaphylococcal activity against methicillin-susceptible and -resistant Staphylococcus aureus as well as metabolite profiling of manuka honey, before and after in vitro simulated gastric (GD), and gastroduodenal digestion (GDD) were investigated.

Undigested manuka honey showed antibacterial activity against all the tested strains. GD sample showed no activity against S. aureus, while GDD honey showed an antistaphyloccoccal activity which was slightly reduced in comparison with undigested sample. To explain these results, methylglyoxal (MGO), to which is ascribed most of the antibacterial activity of MH, was submitted to in vitro simulated GD and GDD.

After digestion, MGO showed antibacterial activity at concentrations definitively higher than those registered in digested MH samples. This results showed that the antistaphylococcal activity registered after digestion cannot be ascribed to MGO. Thus, the metabolite analysis, carried out using an explorative untargeted NMR-based approach and a targeted RP-HPLC-PAD-ESI-MSn analysis focused on bio-active substances, was use to highlight the chemical modifications occurring after digestion.

The results showed that 1) the level of MGO decreases, 2) the content of aromatic compounds, such as leptosin and methyl syringate, markers of manuka honey, was stable under gastric and gastroduodenal conditions, whereas 3) the levels of acetic and lactic acids increase especially after gastroduodenal digestion.

Overall, the results obtained from chemical analysis provide the explanation of the registered antibacterial activity observed after gastroduodenal digestion.

Brazilian Green Propolis Does Not Harm Genetic Material

Evaluation of the genotoxicity/mutagenicity and antigenotoxicity/antimutagenicity induced by propolis and Baccharis dracunculifolia, by in vitro study with HTC cells

Toxicol In Vitro. 2016 Feb 15. pii: S0887-2333(16)30020-0

The ethanolic extract of propolis, especially the Brazilian green type, is widely and mainly used for therapeutic purposes despite the lack of knowledge about its effects and its cellular mode of action.

This type of propolis, derived from Baccharis dracunculifolia (alecrim-do-campo), has been extensively commercialized and the consumers use it to enhance health. This work aimed to assess the genotoxic/mutagenic and antigenotoxic/antimutagenic potentials of the ethanolic extracts of Brazilian green propolis and of B. dracunculifolia, on mammalian cells. It was not observed genotoxic and mutagenic effects by both extracts.

After evaluate the exposure of the cells to each extract with a recognized mutagen, simultaneously, the results showed a significant reduction on DNA damage. The experiment carried out with a pre-incubation period was more effective than without incubation test, showing that the tested extracts were able to inactivate the mutagen before it could react with the DNA.

Manuka Honey Chemical Markers Identified

Manuka honey put under the microscope

By David Porter
Bay of Plenty Times

Scientists have completed a four-year research programme to identify the key chemical markers of manuka honey.

The outcomes will boost long-term consumer confidence in the high-value export product, say experts associated with the Manuka Honey ID Project.

The project is a collaboration between the Unique Manuka Factor Honey Association (UMFHA), major Bay of Plenty-based honey and health products company Comvita, and Hamilton-based Analytica Laboratories.

Data sets from the research are currently with Dr Adrian Charlton of UK food testing and research agency Fera for peer review, and were expected back within the next week or so...

Analysis of Phenolic Content in Korean Propolis

Relationship between total phenolic contents and biological properties of propolis from 20 different regions in South Korea

BMC Complement Altern Med. 2016 Feb 18;16(1):65

BACKGROUND:

Propolis (or bee glue), collected from botanical sources by honey bee, has been used as a popular natural remedies in folk medicine throughout the world. This study was conducted to assess growth inhibitory effects of ethanol extracts of propolis (EEPs) from 20 different regions in South Korea on human intestinal bacteria as well as their human β-amyloid precursor cleavage enzyme (BACE-1), acetylcholinesterase (AChE) inhibitory, antioxidant, antiproliferative, and anti-human rhinovirus activities.

METHODS:

The Bonferroni multiple-comparison method was used to test for significant differences in total polyphenol and flavonoid contents among EEP samples using SAS 9.13 program. Correlation coefficient (r) analysis of the biological activities of EEP samples was determined using their 50 % inhibition concentration or minimal inhibitory concentration values and their polyphenol or flavonoid contents in 20 native Korean EEP samples.

RESULTS:

The amounts of total polyphenol and flavonoids in the Korean EEP samples ranged from 49 to 239 mg gallic acid equivalent (GAE)/g EEP (Brazilian, Chinese, and Australian samples, 127-142 mg GAE/g EEP) and from 21 to 50 mg quercetin equivalent (QE)/g EEP (Brazilian, Chinese, and Australian samples, 33-53 mg QE/g EEP), respectively. Correlation coefficient analysis showed that total polyphenol contents may be negatively correlated with 2,2-diphenyl-1-picrylhydrazyl free radical scavenging activity (r = -0.872) and total flavonoid content has no correlation with the activity (r = 0.071). No direct correlation between BACE-1 inhibition, AChE inhibition, or antiproliferative activity and total polyphenol or total flavonoid content in Korean EEP samples was found. Gram-positive and Gram-negative bacteria were observed to have different degrees of antimicrobial susceptibility to the EEP samples examined, although ciprofloxacin susceptibility among the bacterial groups did not differ greatly.

CONCLUSIONS:

Further studies will warrant possible applications of propolis as potential therapeutic BACE-1 blocker, antioxidant, antiproliferative agent, and antimicrobial agent.

Manuka Honey Business Abuzz with Activity

Wednesday, 17 February 2016

Competition is hotting up among exporters to secure access to precious manuka honey supply, reports KPMG New Zealand.

Keith Richards, from KPMG’s Hamilton office, says high export returns and a limited supply of precious manuka honey is driving a “scramble” to secure access to both hives and manuka-producing land.

“Manuka honey is a fairly scarce commodity, and the issue that many processors and exporters are facing is the ability to guarantee their supply,” he says...

Surgihoney Effective at Preventing Bioflms from Forming

In vitro activity of an engineered honey, medical-grade honeys, and antimicrobial wound dressings against biofilm-producing clinical bacterial isolates

J Wound Care. 2016 Feb;25(2):93-102

OBJECTIVE:

Honey is recognised to be a good topical wound care agent owing to a broad-spectrum of antimicrobial activity combined with healing properties. Surgihoney RO (SH1) is a product based on honey that is engineered to produce enhanced reactive oxygen species (ROS) and has been reported to be highly antimicrobial. The objective was to investigate the ability of the engineered honey and its comparators to prevent biofilm formation in vitro.

METHOD:

We tested the ability of three medical-grade honeys SH1, Activon manuka honey (MH) and Medihoney manuka honey (Med), alongside five antimicrobial dressings (AMDs) to prevent the formation of biofilms by 16 isolates. Honeys were serially double diluted from 1:3 down to 1:6144 and the lowest dilution achieving a statistically significant reduction in biomass of at least 50%, compared with untreated controls, was recorded.

RESULTS:

Although all the honeys were antibacterial and were able to prevent the formation of biofilms, SH1 was the most potent, with efficacy at lower dilutions than the medical honeys for five isolates, and equivalent dilutions for a further six. Additionally, SH1 was superior in antibacterial potency to three commercially available AMDs that contain honey.

CONCLUSION:

SH1 is effective at preventing bioflms from forming and is superior to medical honeys and AMDs in in vitro tests.

Bee Venom Component Inhibits Tumor Cell Growth

Melittin suppresses cathepsin S-induced invasion and angiogenesis via blocking of the VEGF-A/VEGFR-2/MEK1/ERK1/2 pathway in human hepatocellular carcinoma

Oncol Lett. 2016 Jan;11(1):610-618

Melittin, a significant constituent of Apis mellifera (honeybee) venom, is a water-soluble toxic peptide that has traditionally been used as an antitumor agent. However, the underlying mechanisms by which it inhibits tumor cell growth and angiogenesis remain to be elucidated.

In the present study, screening for increased cathepsin S (Cat S) expression levels was performed in MHCC97-H cells and various other hepatocellular carcinoma cell lines by reverse transcription-polymerase chain reaction and western blot analysis. A pcDNA3.1-small hairpin RNA (shRNA)-Cat S vector was stably transfected into MHCC97-H cells (shRNA/MHCC97-H) in order to knockdown the expression of Cat S. The effects resulting from the inhibition of Cat S-induced proliferation, invasion and angiogenesis by melittin were examined using cell proliferation, cell viability, flat plate colony formation, migration, wound healing, Transwell migration and ELISA assays.

In order to substantiate the evidence for melittin-mediated inhibition of Cat S-induced angiogenesis, Cat S RNA was transfected into primary human umbilical vein endothelial cells (Cat S-HUVECs) to induce overexpression of the Cat S gene. The effects of melittin on HUVECs were examined using Transwell migration and tube formation assays.

The findings demonstrated that melittin was able to significantly suppress MHCC97-H cell (Mock/MHCC97-H) proliferation, invasion and angiogenesis, as well as capillary tube formation of Cat S-HUVECs, in a dose-dependent manner. However, proliferation, invasion and angiogenesis in shRNA/MHCC97-H and in native HUVECs (Mock-HUVECs) were unaffected. In addition, melittin specifically decreased the expression of phosphorylated (activated) Cat S, and components of the vascular endothelial growth factor (VEGF)-A/VEGF receptor 2 (VEGFR-2)/mitogen-activated protein kinase kinase 1 (MEK1)/extracellular signal-regulated kinase (ERK)1/2 signaling pathway in Mock/MHCC97-H cells.

In conclusion, the inhibition of tumor cell growth and anti-angiogenic activity exerted by melittin may be associated with anti-Cat S actions, via the inhibition of VEGF-A/VEGFR-2/MEK1/ERK1/2 signaling.

Stingless Bee Honeys Effective Against Multidrug-Resistant Bacteria

Antibacterial synergic effect of honey from two stingless bees: Scaptotrigona bipunctata Lepeletier, 1836, and S. postica Latreille, 1807

Sci Rep. 2016 Feb 12;6:21641

Several studies have tested antimicrobial activity of combinations of honey and various substances.

In this study, we tested a combination of two stingless bee honeys against various bacterial strains. In particular: the antibacterial activity of honeys produced by Scaptotrigona bipunctata (SB) and Scaptotrigona postica (SP) was evaluated against Gram-positive and Gram-negative bacterial strains by agar well diffusion assays, minimum inhibitory concentration (MIC) assessment, construction of growth and viability curves and scanning electron microscopy (SEM).

The interaction of the two honeys was also evaluated by the checkerboard assay. Inhibition zones ranged from 8 to 22 mm. The MIC values of the individual honeys ranged from 0.62 to 10% (v v(-1)) and decreased to 1/4 to 1/32 when the honeys were combined. SEM images showed division inhibition and cell wall disruption for the SB and SP honeys, respectively, and these alterations were observed in same field when the SB and SP honeys were combined.

This study demonstrated that the natural honeys possess in vitro antimicrobial activity against Gram-positive and Gram-negative bacteria, including multidrug-resistant strains. Combination of the SB and SP honeys could lead to the development of new broad-spectrum antimicrobials that have the potential to prevent the emergence of resistant bacterial strains.