• International Journal of Industrial Entomology and Biomaterials
• /
• 제14권2호
• /
• pp.137-142
• /
• 2007

The antibacterial activity of Korean honey bee venom (KBV) was examined against seven major bacterial mastitis pathogens, Enterococcus faecium, Escherichia coli, Methicillin resistant Staphylococcus aureus (MRSA), Staphylococcus aureus, Streptococcus intermedius, Streptococcus oralis and Streptococcus uberis isolated from infected mammary quarters of cows. Seven bacterial mastitis pathogens were studied for antibacterial activity of the KBV by disc diffusion assay, minimal inhibitory concentrations (MIC) and bacterial count in milk samples. The KBV showed activity against Ent. faecium, E. coli, MRSA, Staph. aureus, Strep. intermedius. The order of susceptibility of the bacteria against the KBV was Staph. aureus>MRSA>E. coli>Strep. intermedius>Ent. faecium>Strep. oralis>Strep. uberis. The MIC against Staph. aureus, MRSA and E. coli were stronger effect as compared with standard drug. The effect of the KBV ($100{\mu}g\;ml^{-1}$) on the viability of Ent. faecium, E. coli, MRSA, Staph. aureus, Strep. intermedius, Strep. oralis and Strep. uberis in milk differed significantly with each other within 12 h incubation period. The results indicate that KBV has significant antibacterial effects against major bacterial mastitis bacteria, Ent. faecium, E.coli, MRSA, Staph. aureus, Strep. intermedius. Results of the study indicate the potential use of KBV as alternative to antibiotic therapy. Further investigations are needed though to confirm its efficacy and its effects on the animals.

• Journal of Microbiology and Biotechnology
• /
• 제27권1호
• /
• pp.84-91
• /
• 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.

• 분석과학
• /
• 제27권6호
• /
• pp.277-283
• /
• 2014

한외여과법을 이용하여 봉독의 알러지 원인성분인 $PLA_2$를 제거하기 위하여 압력, 농도, 분자크기의 영향을 조사하였다. 봉독의 주요성분 분자량을 바탕으로 한외여과막의 투과크기를 선정하고 농도와 압력을 달리하였다. 그 결과, melittin과 apamin 함량은 유지되면서 $PLA_2$를 제거하는 최적조건(1 mg/mL, 20 psi, 10,000 dalton)을 찾았으며, 이를 HPLC와 SDS-PAGE로 확인하였다.

• BMB Reports
• /
• 제53권8호
• /
• pp.419-424
• /
• 2020

Bee venom (BV), secreted from the venom gland of the honey bee, contains several biological active compounds. BV has been widely used as a traditional medicine for treating human disease, including cancer. In this study, we have shown the molecular mechanism underlying the therapeutic effect of BV on cancer. Treatment with BV reduced the proliferation of cervical-cancer cells in a dose- and time-dependent manner. Interestingly, the killing effect of BV was specific to HPV-positive cervical-cancer cell lines, such as Caski and HeLa cells, and not to HPV-negative cervical-cancer cells (C33A). BV reduced the expression of HPV E6 and E7 at RNA and protein levels, leading to an increase in the expression of p53 and Rb in Caski and HeLa cells. Further, BV decreased the levels of cell-cycle proteins, such as cyclin A and B, and increased the levels of cell-cycle inhibitors, such as p21 and p27. BV significantly induced apoptosis and inhibited wound healing and migration of cervical-cancer cells. It also upregulated the expression of pro-apoptotic BAX and downregulated the expression of anti-apoptotic Bcl-2 and Bcl-XL. Cleavage of caspase-3, caspase-9, and PARP were also induced by BV treatment, whereas the phosphorylation of mitogenic signaling-related proteins, such as AKT, JNK, p38, and ERK, were downregulated. Our results indicate that BV has a therapeutic selectivity for HPV-positive malignant cells, so further clinical studies are needed to assess its clinical application.

• 대한약침학회지
• /
• 제14권3호
• /
• pp.19-45
• /
• 2011

Objectives: This study was performed to analyse the effects of Sweet Bee Venom(Sweet BV-pure melittin, the major component of honey bee venom) on the central nervous 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, Sweet BV was administered in thigh muscle of rats. And checked the effects of Sweet BV on the central nervous system using the functional observational battery (FOB), which is a neuro-toxicity screening assay composed of 30 descriptive, scalar, binary, and continuous endpoints. And home cage observations, home cage removal and handling, open field activity, sensorimotor reflex test/physiological measurements were conducted. Results: 1. In the home cage observation, there was not observed any abnormal signs in rats. 2. In the observation of open field activity, the reduction of number of unit areas crossed and rearing count was observed caused by Sweet BV treatment. 3. In the observation of handling reactivity, there was not observed any abnormal signs in rats. 4. In the observation of sensorimotor reflex tests/physiological measurements, there was not observed any neurotoxic signs in rats. 5. In the measurement of rectal temperature, treatment of Sweet BV did not showed great influences in the body temperature of rats. Conclusions: Above findings suggest that Sweet BV is relatively safe treatment in the central nervous system. But in the using of over dose, Sweet BV may the cause of local pain and disturbance of movement. Further studies on the subject should be conducted to yield more concrete evidences.

• 대한약침학회지
• /
• 제23권4호
• /
• pp.212-219
• /
• 2020

Objectives: Nowadays cancer treatment is an important challenge in the medical world that needs better therapies. Many active secretions produced by insects such as honey bees used to discover new anticancer drugs. Bee venom (BV) has a potent anti inflammatory, anti cancer and tumor effects. The aim of present study is evaluation of anticancer effects induced by Apis mellifera venom (AmV) on cell Lines. Methods: AmV was selected for study on cancer cell lines. Total protein, molecular weight and LD50 of crude venom were determined. Then, cells were grown in Dulbecco's Modified Eagle medium supplemented with 10% fetal bovine serum and 1% antibiotics. The A549, HeLa and MDA-MB-231 cell Lines were exposed by different concentration of AmV. The morphology of cells was determined and cell viability was studed by MTT assay. Evaluation of cell death was determined by and DNA fragmentation. Results: The results from MTT assay showed that 3.125 ㎍/mL of A549, 12.5 for HeLa and 6.25 ㎍/mL of MDA-MB-231 killed 50% of cells (p < 0.05). Morphological analysis and the results from hoescht staining and DNA fragmentation indicated that cell death induced by AmV was significantly apoptosis. Conclusion: The data showed that using lower dosage of AmV during treatment period cause inhibition of proliferation in time and dose dependant manner. Findings indicated that some ingredients of AmV have anticancer effects and with further investigation it can be used in production of anticancer drugs.

• 한국임상수의학회지
• /
• 제16권1호
• /
• pp.155-162
• /
• 1999

This study was performed to assess that clinco-therapeutic effect of natural Italian honeybee (Apis mellifera) venom in adjuvant-induced arthritic rat. Ninety Sprague- Dawley rats of male were injected with complete Freund's adjuvant (CFA). Adjuvant arthritis was produced by a single subcutaneous injection of 1 mg Mycobacterium butyricum suspended in 0.1 ml paraffin oil into the right hindpaw. Righting reflex was uniformly lost and considered to be the point of arthritis development on day 14 after CFA injection. Experimental groups were divided into three groups. When arthritis was developed in the rat hind-paw, tested groups were administrated with prednisolone (10 mg/kg, p.o) and honeybee venom (one bee, s.c) at an interval of two days. Control group was subcutaneously injected with 0.1 ml of physiological saline solution in the rat at an interval of two days. Clinical findings, hematological values and histopathological findings were observed during or after the drugs administration. In tested groups, the development of inflammatory edema and polyarthritis on day 14 after treatment was suppressed. No significant differences of hindpaw edema volume and lameness score between prednisolone and honeybee venom groups were observed during or after therapeutic drugs treatment. WBC counts of prednisolone and honeybee venom treatment groups as compared with the control group were getting remarkably decreased during or after the therapeutic drugs administration(p＜0.01). Erosions of articular cartilage and inflammatory cell infiltrations during or after the therapeutic drugs treatment was effectively suppressed in natural honey venom.

• 분석과학
• /
• 제29권4호
• /
• pp.186-193
• /
• 2016

본 연구는 용규전초로부터 열수추출방법(SNL-W)으로 1차 추출하고 부틸알코올을 이용하여 분별추출을 통해 유기용매분획(SNL-W/B)과 수상분획(SNL-W/W)을 각각 획득하였다. 각 분획의 총 알칼로이드함량과 사포닌함량을 측정하였고 수상분획의 메탄올 용출물은 알칼로이드 성분으로 확인되었다. 꿀벌봉독을 쥐의 복막 비만세포에 처리하여 알러지 유발물질인 히스타민을 분비시키고, 용규의 알칼로이드성분으로 히스타민 분비에 대한 저해효과를 고찰하였다. 그 결과, 용규의 수용성 알칼로이드성분(SNL-W/W/M)은 꿀벌봉독으로 유발된 비만세포의 히스타민 분비에 대하여 강한 저해요인으로 작용하는 것을 확인하였다.

• Journal of Acupuncture Research
• /
• 제22권3호
• /
• pp.37-51
• /
• 2005

비만세포는 염증 및 알레르기 반응과 관련하여 우리 몸에서 주요한 작용을 하는 세포이다. 봉독은 현재까지 진통기전에 관련된 모델로 연구가 진행되어 왔으나, 최근에는 항염증이나 항알러지반응 둥에서 면역세포와 관련 한 연구가 진행 중에 있다. 본 연구는 봉독의 주요성분인 melittin과 MCD Peptide가 비만세포주에서의 유전자 발현에 미치는 영향을 연구함으로써 향후 유전자 언구에 관련한 기초를 제시하고자 하였다. 본 연구에서는 사람의 비만세포주를 이용하여, 세포독성 실험을 거쳐서 얻은 유효농도에서 각각 melittin과 MCD Peptide를 처치하고, 이때 변화하는 유전자의 발현양상을 microarray분석기법을 통하여 정보를 얻었다. 실험적 통계에 의하여 global M이 1 또는 -1 이상인 것을 유의한 것으로 보았을 때, melittin에서는 모두 7개 의 유전자가 항진되고, 8개의 유전자가 어제되었다. MCDP에서는 7개의 유전자가 항진되고 17개의 유전자가 억제되었다. 이들 유전자들이 주로 관련하는 체내의 작용은 세포내에서 단백결합, lymphocyte 기능의 활성화, macrophage 항원관련 및 세포핵의 수용체, GABA A receptor 관련물질, cAMP 반응요소와 연관된 단백질, 보체계 8번 및 B-cell 관련물질, 다낭성 신질환에 관련된 단백물질, 염증관련물질, 혈액응고에 영향을 주는 단백물질등과 연관이 되었다. 이러한 분석결과를 통하여 동복에서의 주요약리작용을 담당하는 melittin과 MCD peptide의 작용기전을 밝히는데 보다 유용한 자료를 얻을 수 있었으며, 향후에 봉독의 주요성분 및 전체봉 독액이 항알레르기반응이나 항염증작용에 미치는 영향에 대한 심도있는 연구가 필요할 것으로 사료된다.