• Korean Journal of Pharmacognosy
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• v.46 no.3
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• pp.203-207
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• 2015

This study was performed to examine the antigenic potential of purified bee venom (Apis mellifera L., PBV) collected using bee venom collector. Antigenic potential of PBV was examined by active systemic anaphylaxis (ASA) in guinea pigs. PBV was subcutaneously administered at 0.025 and 0.05 mg/kg and also as a suspension with adjuvant (Freund's complete adjuvant, FCA). Ovalbumin (OVA) as a suspension with adjuvant was used to introduce positive control response. In the weight measurement and clinical observation, experimental groups didn't show any significant changes compared with control group. In the autopsy of body, the abnormalities of lung were detected only in the positive control. In the ASA test, experimental groups didn't show any symptoms of anaphylaxis like piloerection, hyperpnea and staggering gait. These results suggested that PBV didn't have antigenic potential in guinea pig.

• Journal of Apiculture
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• v.35 no.1
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• pp.49-54
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• 2020

Bee (Apis mellifera L.) venom is used for the treatment of various human diseases due to its known anti-inflammatory and antibacterial properties. This study investigated the effect of purified bee venom (PBV) on adipogenesis in 3T3-L1 preadipocytes. There was no cytotoxicity while cells were treated with PBV by various concentrations. In the PBV treated cells increases in fat storage were inhibited and also confirmed by oil red o staining. To understand the underlying mechanism at the molecular level were examined on the expression of the genes involved in adipogenesis by using real-time PCR. In this cell model, the mRNA level of adipogenic genes such as peroxisome-proliferator-activated receptors gamma (PPARγ) and CAAAT/enhancer binding protein alpha(C/EBPα) were decreased by PAE treatment, comparing with those of control group. Theses results suggest that PBV inhibits adipocyte differentiation in 3T3-L1 cells and can be used as an efficient natural substance to manage anti-obesity.

• Journal of Pharmacopuncture
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• v.18 no.4
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• pp.7-11
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• 2015

Objectives: Bee venom (BV) is a complex mixture of proteins and contains proteins such as phospholipase and melittin, which have an effect on blood clotting and blood clots. The mechanism of action of honey bee venom (HBV, Apis mellifera) on human plasma proteins and its anti-thrombotic effect were studied. The purpose of this study was to investigate the anti-coagulation effect of BV and its effects on blood coagulation and purification. Methods: Crude venom obtained from Apis mellifera was selected. The anti-coagulation factor of the crude venom from this species was purified by using gel filtration chromatography (sephadex G-50), and the molecular weights of the anti-coagulants in this venom estimated by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Blood samples were obtained from 10 rabbits, and the prothrombin time (PT) and the partial thromboplastin time (PTT) tests were conducted. The approximate lethal dose (LD) values of BV were determined. Results: Crude BV increased the blood clotting time. For BV concentrations from 1 to 4 mg/mL, clotting was not observed even at more than 300 seconds, standard deviations $(SDs)={\pm}0.71$; however, clotting was observed in the control group 13.8 s, $SDs={\pm}0.52$. Thus, BV can be considered as containing anti-coagulation factors. Crude BV is composed 4 protein bands with molecular weights of 3, 15, 20 and 41 kilodalton (kDa), respectively. The $LD_{50}$ of the crude BV was found to be $177.8{\mu}g/mouse$. Conclusion: BV contains anti-coagulation factors. The fraction extracted from the Iranian bees contains proteins that are similar to anti-coagulation proteins, such as phospholipase $A_2(PLA_2)$ and melittin, and that can increase the blood clotting times in vitro.

• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.84-91
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• 2017

The concern over the use of melittin in honey bee venom due to its adverse reaction caused by allergens such as phospholipase A2 ($PLA_2$) and hyaluronidase (HYA) has been an obstacle towards its usage. We developed a novel single-step method for melittin purification and the removal of $PLA_2$ and HYA. This study explores the influence of pH, buffer compositions, salt concentration, and types of cation-exchange chromatography resins on the recovery of melittin and the removal of both HYA and $PLA_2$. Melittin was readily purified with a strong cation-exchange resin at pH 6.0 with sodium phosphate buffer. It resulted in a recovery yield of melittin up to 93% (5.87 mg from a total of 6.32 mg of initial melittin in crude bee venom), which is higher than any previously reported studies on melittin purification. $PLA_2$ (99%) and HYA (96%) were also successfully removed. Our study generates a single-step purification method for melittin with a high removal rate of $PLA_2$ and HYA, enabling melittin to be fully utilized for its therapeutic purposes.

• 대한임상약리학회:학술대회논문집
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• 1994.12a
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• pp.30-30
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• 1994

• YAKHAK HOEJI
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• v.56 no.4
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• pp.254-259
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• 2012

To further access honeybee (Apis mellifera L.) venom (BV) as a cosmetic ingredient and potential external treatment for topical use, we investigated its ability to inhibit tyrosinase activity and melanin biosynthesis on melanogenesis in B16F1 melanoma cells. We found that BV increased the cell viability in B16F1 melanoma cell and BV (0.01~1 ${\mu}g/ml$) inhibited melanin synthesis in with 10 nM ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH) for 48 h. In addition, we used reverse transcription-polymerase chain reaction and western blotting for me melanogenesis-related genes such as tyrosinase to examine the mechanisms underlying the inhibitory effects of BV on melanogensis. BV inhibited direct tyrosinase activity, which decreased melanin synthesis in ${\alpha}$-MSH stimulated B16F1 melanoma cells. Thease findings suggest that BV induces the downregulation of melanogenesis by inhibiting tyrosinase activation.

• Korean Journal of Veterinary Service
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• v.31 no.2
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• pp.229-237
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• 2008

This study was conducted to evaluate the effect of honeybee venom, purified using bee venom collector, and feeding of probiotics on the body weight gain, growth rate and hematological characteristics of pigs. A total of 120 pigs were examined and divided into 4 groups 1) Control (basal diet), 2) BV (basal diet + bee venom), 3) PB (basal diet + probiotics), 4) BVPB (basal diet + BV + PB). Average daily weight gain improved significantly in all test groups, especially BVPB (P < 0.05) compared to the controls. There was a significant difference in the feed conversion rate (P < 0.05) and efficiency (P < 0.05) between BVPB and control pigs. Weight gain and survivability was higher in the tests than the controls, but white blood cell count was not. Serum total protein, albumin and IgG concentration of BVPB were slightly higher than those of controls. These results suggest that treated honeybee venom and probiotics should be used together to effectively increase the productivity of pigs.

• Korean Journal of Pharmacognosy
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• v.43 no.2
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• pp.167-172
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• 2012

Osteoarthritis (OA) is an age-related joint disease characterized by degradation of articular cartilage and its association with chronic pain. Purified Bee venom (PBV) has been traditionally used to the treatment for inflammatory diseases. The purpose of the current study is to examine whether PBV regulates the pro-inflammation against a mouse model of knee OA induced by papain. We studied the effect of PBV on papain-induced OA in the knee joints of mice. Mice were split into following groups: normal control, papain induced OA, OA treated with PBV, OA treated with meloxicam as positive control. Proteoglycan deposition was analyzed by safranin O-fast green staining and H&E staining. Papain injection significantly degraded the proteoglycan in OA mice at 42 days. Cartilage proteoglycan density was significantly higher in PBV treated OA group than those of the positive control groups. These results demonstrate that PBV efficiently suppresses pathological processes in an OA model. Thus, PBV could be a potential therapeutic strategy for the treatment of OA.

• Journal of Pharmacopuncture
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• v.25 no.4
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• pp.390-395
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• 2022

Objectives: Sweet bee venom (sBV) is purified from Apis mellifera, containing a high level of melittin-its main component. It has been used as a therapeutic agent for pain relief and anti-inflammation, as well as for treating neuronal abnormalities. Recently, there have been studies on the therapeutic application of sBV for anticancer treatment. In the present study, we investigated the pharmacological effect of sBV treatment in A549 human lung cancer cells. Methods: We used microscopic analysis to observe the morphological changes in A549 cells after sBV treatment. The MTT assay was used to examine the cytotoxic effect after dose-dependent sBV treatment. Molecular changes in sBV were evaluated by the expression of apoptosis marker proteins using western blot analysis. Results: Microscopic analysis suggested that the growth inhibitory effect occurred in a dose-dependent manner; however, cell lysis occurred at a concentration over 20 ㎍/mL of sBV. The MTT assay indicated that sBV treatment exhibited a growth inhibitory effect at a concentration over 5 ㎍/mL. On fluorescence activated cell sorting analysis, G0 dead cells were observed after G1 arrest at treatment concentrations up to 10 ㎍/mL. However, rapid cell rupture was observed at a concentration of 20 ㎍/mL. Western blot analysis demonstrated that sBV treatment modulated the expression of multiple cell death-related proteins, including cleaved-PARP, cleaved-caspase 9, p53, Bcl2, and Bax. Conclusion: sBV induced cell death in A549 human lung cancer cells at a pharmacological concentration, albeit causing hemolytic cell death at a high concentration.

• Journal of Sericultural and Entomological Science
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• v.52 no.1
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• pp.6-9
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• 2014

This study was for the investigation of the stability of purified bee venom (PBV) during the treatment in the pH range from pH2 to pH9 for 24 hours, respectively. Changes of components and physiological functionalities in PBV were by evaluated silver staining, and melittin contents were measured by liquid chromatography. The antimicrobial activity against bacteria by minimum inhibitory concentration (MIC) and effect of the cell regeneration were measured by 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl tetrazolium bromide (MTT) assay using human dermal fibroblast (HDF) cell. The main proteins such as melittin and phospholipase $A_2$ showed no characteristic changes. The antimicrobial activity and effect of cell regeneration showed no difference from pH2 to pH9. From this study, we suggest that components and physiological functionalities of PBV against treated pH were kept stability at from pH2 to pH9.