• 한국동물위생학회지
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• 제41권3호
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• pp.171-177
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• 2018

Purified bee venom was collected from colonies of honeybees (Apis mellifera L.) using a bee venom collector under sterile conditions and then purified under strict laboratory conditions. Purified bee venom contained $63.9{\pm}5.4%$ melittin, $10.9{\pm}1.6%$ phospholipase A2, and $2.3{\pm}0.3%$ apamin. Purified bee venom has various anti-bacterial, anti-inflammatory and immunostimulating effects. In this study, we evaluated purified bee venom which are mammary gland cells, MAC-T cells are used to increase the synthesis of milk protein. Purified bee venom promoted the proliferation of MAC-T cells at concentrations below $1{\mu}g/mL$, but cytotoxicity at $10{\mu}g/mL$ and above. As a result of the increase in the synthesis of ${\beta}-casein$, a milk protein after treatment with MAC-T cells at a concentration of the bee venom without cytotoxicity, the ${\beta}-casein$ content in the cell culture was increased when treated at a concentration of 1 ng/mL or more. In addition, it was confirmed that purified bee venom significantly increased the expression of bovine ${\beta}-casein$ (bCSNB) mRNA, a ${\beta}-casein$ synthesis gene, at a concentration of 1 ng/mL or more. These results suggest that purified bee venom can be used to increase the production of livestock by ultimately increasing the expression of milk protein.

• 생약학회지
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• 제47권1호
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• pp.43-48
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• 2016

The aim of the study was performed to examine the anticariogenic potential of purified bee venom (Apis mellifera L., PBV) collected using bee venom collector from cariogenic bacteria, Streptococcus mutans, Streptococcus sanguis, Porphyromonas gingivalis, and Fusobacterium nucleatum. The anticariogenic effect of purified bee venom was evaluated by agar well diffusion method, minimum inhibitory concentraion (MIC), minimum bactericidal concentration (MBC), and postantibiotic effect (PAE). The human lower gingiva epithelial cell cytotoxicity of purified bee venom was also evaluated. Purified bee venom exhibited significant inhibition of bacterial growth of S. mutans, S. sanguis, P. gingivalis, and F. nucleatum with MIC value of 0.68, 0.85, 3.49, and $2.79{\mu}g/ml$, respectively. The MBC value of purified bee venom against S. mutans, S. sanguis, P. gingivalis, and F. nucleatum was 1.34, 1.67, 8.5, and $6.8{\mu}g/ml$. Furthermore, the results of PAE values against S. mutans, S. sanguis, P. gingivalis, and F. nucleatum showed the bacterial effect with 3.3, 3.45, 2.0, and 2.0. The concentration below 1 mg/ml of purified bee venom had no cytotoxicity in the human lower gingiva epithelial cell. These results suggested that purified bee venom have great potenial as anticariogenic agents.

• 한국동물위생학회지
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• 제39권3호
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• pp.159-166
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• 2016

Clostridium perfringens, Salmonella thyphimurium and S. Montevideo isolated from the intestines of dead broiler chickens in Korea were tested for antibacterial effects to purifed bee venom. Purified bee venom from Apis mellifera L. has been used as natural antimicrobial compounds in pigs, cows, dairy cattle and chicken farms in Korea. To investigate antibacterial effect of purified bee venom was evaluated by agar well diffusion method, minimum inhibitory concentraion (MIC), minimum bactericidal concentration (MBC), and postantibiotic effect (PAE). Purified bee venom exhibited significant inhibition of bacterial growth of C. perfringens, S. thyphimurium and S. Montevideo with MIC value of 0.85, 0.68 and $0.69{\mu}g/mL$, respectively. The MBC value of purified bee venom against C. perfringens, S. thyphimurium and S. Montevideo were 3.33, 2.66 and $2.86{\mu}g/mL$. Furthermore, the results of PAE values against C. perfringens, S. thyphimurium and S. Montevideo showed the bacterial effect with 3.5, 4.0 and 3.5 hr. Stability of pufifed bee venom at acidity from pH 1 to pH 8 for 24 hr was the antibacterial activity for C. perfringens, S. thyphimurium and S. Montevideo and melittin contents. Also purified bee venom processed through the heating for 15 min, there was no signification loss of the antibacterial activity and melittin at below $100^{\circ}C$. These results obtained in this study suggest that purified bee venom might be utilized as a feed additive in poultry diets.

• Journal of Acupuncture Research
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• 제19권2호
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• pp.28-38
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• 2002

Objective : This study is aimed to investigate the effects of bee venom and purified bee venom on cell death in synovial cell line. Methods : It was evaluated by using MTT assay, morphological method, flow cytometry, immunocytochemistry analysis, RT-PCR. Results : The result obtained is as follows. 1. The MTT assay demonstrated that synovial cell viability was significantly inhibitted dose-dependently by treatment with BV and PBV in comparison with control. And the inhibitory effect of BV and PBV was almost same. 2. The morphologic study demonstrated that synovial cell showed apoptotic body resulted from apoptosis after treatment with BV and PBV for 6 hours using microscope. 3. The Flow cytometry demonstrated that apoptosis of synovial cell treated with BV and PBV was related with stop of cell cycle in stage of G0/G1. 4. Immunocytochemistry assay demonstrated that COX-II and iNOS were slightly expressed by treatment with BV and PBV in comparison with control group. 5. RT-PCR analysis demonstrated that COX-II were almost down-regulated by high dose treatment with BV and PBV in comparison with control group. iNOS were well down-regulated by treatment with $5{\mu}g/ml$ BV and PBV whereas it was well expressed in control group. Conclusion : These results suggest that bee venom and purified bee venom have significant effect on cell death in synovial cell line and further study is needed in vivo.

• 분석과학
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• 제29권4호
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• pp.194-201
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• 2016

본 연구는 수용액 상태에서 정제봉독 (purified bee venom, PBV)과 봉독멜리틴 (bee venom melittin, BVM)을 보관하기 위하여 온도, 용매, 농도의 조건에 따른 안정성을 비교 실험한 결과이다. Prep-HPLC (column: C₄)를 이용하여 정제봉독으로부터 98.2%인 봉독멜리틴을 분리하여 사용하였다. 용매에 따른 안정성 실험에서, 생리식염수의 경우 정제봉독은 안정성이 증가되고 봉독멜리틴은 감소되었다. 증류수의 조건에서 정제봉독의 안정성은 감소되었으나 봉독멜리틴은 증가되는 현상이 확인되었다. 봉독멜리틴의 안정성을 장기간 유지하기 위한 최적조건으로는 보관온도 4 ℃, 증류수, 시료의 농도는 1.0 mg/mL의 결과를 얻었다.

• 한국양봉학회지
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• 제32권3호
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• pp.269-273
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• 2017

본 연구에서는 서양종꿀벌의 일벌에서 채집하여 정제한 정제봉독이 알코올 분해능에 관련이 있는 ADH와 ALDH 효소 활성에 미치는 영향을 알아보고자 하였다. ADH와 ALDH 활성은 in vitro kit를 사용하여 측정하였다. 그 결과 정제봉독 처리에 의해 ADH와 ALDH 효소 활성이 크게 증가하는 것으로 확인되었다. ADH 효소는 1mg/ml 이상의 정제봉독을 처리했을 경우 양성대조구(2mg/ml) 대비 $88.6{\pm}7.34%$의 활성도를 보였으며, ALDH 효소 역시 1mg/ml 이상의 정제봉독에서 양성대조구(2mg/ml) 대비 $94.6{\pm}0.57%$의 활성도를 나타내었다. 이상의 결과로 정제봉독은 숙취 해소능에 영향을 미치는 지표물질인 ADH와 ALDH 효소 활성을 크게 증가시키는 것으로 사료되었으며, 향후 추가시험을 통해 숙취해소 작용을 갖는 식의약품 소재로 사용 될 수 있을 것으로 생각된다.

• 약학회지
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• 제60권2호
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• pp.53-57
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• 2016

This study was performed to examine the skin phototoxicity of purified bee venom (Apis mellifera L.) collected using bee venom collector. To confirm whether the gel containing purified bee venom (BV gel) causes photototoxicity when used for the skin medicinal products, phototoxicity testing was conducted using guinea pig models. The BV gel (0.1 ml/site) was administered transdermally to guinea pigs. 8-MOP was used to introduce positive control response. After administration, the guinea pigs were irradiated with UVA ($15J/cm^2$) with doses based on standard phototoxicity study guidelines. In the weight measurement and clinical observation, BV gel groups didn't show any significant changes compared with control group. BV gel groups did not show any symptoms such as erythema and edema formation of skin. This study demonstrated that BV gel has promising potential external treatment for topical uses that do not induce significant levels of skin phototoxicity.

• 한국식품위생안전성학회지
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• 제32권3호
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• pp.228-233
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• 2017

서양종 꿀벌의 일벌의 독을 채취하여 정제한 정제봉독에 대한 세균에서의 돌연변이 유발성 검색을 위하여 S. typhimurium의 히스티딘 요구성 균주 TA100, TA1535, TA98, 그리고 TA1537의 균주와 대장균 E. coli의 트립토판 요구성 균주인 WP2 uvrA를 이용해 복귀돌연변이시험을 실시하였다. 정제봉독은 부형제로 사용한 멸균생리식염수에 용해되었으며 모든 농도군의 조제물에서 침전은 관찰되지 않았으며, 정제봉독 조제물을 top agar와 혼합할 때 모든 농도군에서 혼탁이나 침전이 관찰되지 않았다. 대사활성계 적용 TA100, TA1535, TA98 균주에 대해 0, 1.5, 5, 15, 50, 150 및 500 µg/plate의 범위를 설정하고 미 적용시엔 0, 0.15, 0.5, 1.5, 5, 15 및 $500{\mu}g/plate$의 범위로 설정하였다. TA1537과 WP2 uvrA균주에 대한 농도 범위은 대사활성계 적용시엔 0, 5, 15, 50, 150, 500 및 1,500를 미 적용시엔 0, 1.5, 5, 15, 50, 150 및 500 µg/plate 범위로 설정하여 시험을 수행하였다. 그 결과 모든 시험균주에서 대사활성계 적용 여부에 상관없이 정제봉독 처리군의 평균 집락 수는 증가를 나타내지 않았으며, 양성판정 기준을 만족시키지 못하였다. 따라서, 정제봉독은 본 시험조건 하에 사용한 시험 균주에 복귀돌연변이를 유발하지 않는 것으로 사료되었다.

• 대한약침학회지
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• 제4권2호
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• pp.5-15
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• 2001

This study was carried out for production of neutral antibody to bee venom $(anti-phospholipase\;A_2IgY)$. Hen layings were injected repeatedly with bee venom and phospholipase $A_2$ with Freund's adjuvant. Specific antibody in egg yolk from immunized hen laying was separated, and purified, also immunological characteristics of anti phospholipase $A_2\;IgY$ was invested. The results were summarized as follows: 1. Phospholipase $A_2$ was showed single band at molecular weight 17,000 in SDS-PAGE and bee venom was showed two band at molecular weight 17,000 and under molecular weight 6,500 in SDS-PAGE. 2. During 70 days after hen immunized with bee venom and phospholipase $A_2$, antibodies(anti-bee venom IgY) to bee venom were showed poor ELISA value in egg yolk, but antibodies$(anti-Phospholipase\;A_2IgY)$ to phospholipase $A_2$ in egg yolk were increased ELISA value from 8 days or 15 days and found maximum ELISA value at 42 days. Also after booster at 49 days, ELISA value of anti Phospholipase $A_2\;IgY$ in egg yolk was supported at optical density(O.D) 1.0 level, continuously. 3. Titer of phospholipase $A_2\;IgY$ was showed 1: 32,000. 4. In double immunodiffusion test to phospholipase $A_2$ after double dilution of anti-phospholipase $A_2\;IgY$, only precipitation line was made in 1:1 dilution well of anti-Phospholipase $A_2\;IgY$. But In immunodiffusion test to anti-phospholipase $A_2\;IgY$ after double dilution of phospholipase $A_2$, Precipitation line to 250ul/ml well of phospholipase $A_2$ was showed. In double immunodiffusion test to bee venom(1mg/ml) after double dilution anti-phospholipase $A_2\;IgY$, all well without 1:32 dilution well were showed strong precipitation line. 5. In dot bloting test to anti-phospholipase $A_2\;IgY$ after diluting bee venom(0.5mg/ml), dot bloting color was showed clearly to $1/100(5{\mu}g/ml)$ in bee venom.

• 대한약침학회지
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• 제14권3호
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• pp.47-53
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• 2011

Objectives: This study was performed to analyse the effects of Sweet Bee Venom(SBV-purified melittin supported by G&V Co., the major component of honey bee venom) on the respiratory system in rats. Methods: All experiments were conducted at Biotoxtech Company, a non-clinical studies authorized institution, under the regulations of Good Laboratory Practice(GLP). Male rats of 5 weeks old were chosen for this study and after confirming condition of rats was stable, SBV was administered in thigh muscle of rats in 0.175, 0.35 and 0.7 mg/kg dosage. And checked the effects of SBV on the respiratory system using the whole body plethysmography. And respiratory rate, tidal volume and minute volume of rats were checked after administered SBV (melittin). Results: 1. In the measurement of respiratory rate, there were not observed any significant differences compared with control group. 2. In the measurement of tidal volume, there was not observed any significant differences compared with control group. 3. In the measurement of minute volume, 0.35mg/kg dosage group showed significant differences compared with control group. But we estimated that this result was caused by individual differences. Conclusions: Above findings suggest that SBV seems to be safe treatment in the respiratory system of rats. And further studies on the subject should be conducted to yield more concrete evidences.