Bee venom, which serves as a weapon to defend the colony from predator attacks, induces an immediate local inflammatory response that causes acute redness and swelling at the site of the sting. This venom-induced inflammation is a rapid anti-predatory defense strategy of the bee against vertebrate predators. Although acute inflammation by venom from venomous arthropods, including bees, is a typical response, how venom acutely elicits inflammatory responses remains unknown. Here, we identify a novel mechanism underlying acute inflammation and provide a rationale for the presence of superoxide dismutase (SOD3) in bee venom. In mouse models, paradoxically, SOD3 in bee venom (bvSOD3) acts as a reactive oxygen species (ROS)-based harm-inducing system to promote acute inflammation. Exogenous bvSOD3 rapidly induced overproduction of H2O2 through endogenously produced superoxide by venom components, such as melittin and phospholipase A2 (PLA2), which then upregulated the expression of proinflammatory genes and promoted the acute inflammatory response. Furthermore, a more severe noxious effect by bvSOD3 elevated a type 2 immune response, and bvSOD3 immunization protected against bvSOD3-mediated toxicity. Our findings that bvSOD3 promotes an acute inflammatory response and induces a protective immune response against inflammation may offer a new approach in venom therapy/immunotherapy.

Bee venom contains a variety of toxic components, including vitellogenin, which display various biological, toxicological,and pharmacological activities. However, the biological actions of vitellogenin, a venom protein in bee venom, remainlargely unknown. Here, we demonstrate that Asiatic honeybee (Apis cerana) venom vitellogenin (AcV-Vg) exhibits anti-oxidantand anti-microbial activities. AcV-Vg is expressed in the venom gland and is then secreted into venom. The recombinantAcV-Vg protein was produced in baculovirus-infected insect cells. We found that AcV-Vg reduced cytotoxicity and oxidativedamage against oxidative stress. Furthermore, AcV-Vg bound to microbial surfaces and induced structural damage in themicrobial cell walls, which, in turn, exhibited anti-microbial activity against bacteria and fungi. Together, our data demonstratedthat the bee venom protein AcV-Vg has multifunctional roles as an anti-oxidant and anti-microbial agent.

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 트립토 판 요구성 균주인 WP2 uvrA를 이용해 복귀돌연변이시험 을 실시하였다. 정제봉독은 부형제로 사용한 멸균생리식 염수에 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 50 μg/plate의 범위로 설 정하였다. 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 처리군 의 평균 집락 수는 증가를 나타내지 않았으며, 양성판정 기준을 만족시키지 못하였다. 따라서, 정제봉독은 본 시험 조건 하에 사용한 시험 균주에 복귀돌연변이를 유발하지 않는 것으로 사료되었다.

Bee venom is a complex mixture of toxic components that induces immediate local inflammatory and allergic responses. However, the presence and role of superoxide dismutase (SOD) in bee venom have not been previously investigated. Here, we provide the first demonstration that bee venom contains Cu,Zn SOD (SOD3), a novel extracellular component that promotes local inflammation. Bee venom SOD3 was shown to be an oxidant, rather than an antioxidant, that induces the inflammation-signaling molecule H2O2 in vivo. H2O2 plays a pathological role by triggering an immediate local inflammatory response. Furthermore, bee venom SOD3 significantly induced the activation of proinflammatory mediators (TNF-α and COX-2) and cytokines (IL-1β and IL-6) via the overproduction of H2O2 in mice. Our data demonstrate that bee venom SOD3 induced H2O2, which drives an immediate local inflammatory response, indicating a novel mechanism underlying bee venom-induced local inflammation.

Bee venom contains a variety of peptide constituents that have various biological, toxicological, and pharmacological actions. However, the biological actions of secapin, a venom peptide in bee venom, remain largely unknown. Here, we provide the first evidence that the Asiatic honeybee (Apis cerana) secapin (AcSecapin-1) exhibits anti-fibrinolytic, anti-elastolytic, and anti-microbial activities. AcSecapin-1 functions as a serine protease inhibitor-like peptide that has inhibitory effects against plasmin, elastases, microbial serine proteases, trypsin, and chymotrypsin. Consistent with these functions, AcSecapin-1 inhibited the plasmin-mediated degradation of fibrin to fibrin degradation products, thus indicating the role of AcSecapin-1 as a clotting factor. AcSecapin-1 also inhibited both human neutrophil and porcine pancreatic elastases. Furthermore, AcSecapin-1 exhibited anti-microbial activity against fungi and Gram-positive and Gram-negative bacteria. Taken together, our data demonstrated that AcSecapin-1 has a multifunctional role as an anti-fibrinolytic agent, an anti-elastolytic agent, and an anti-microbial peptide, and our data suggested novel functions for the biological actions of the bee venom peptide, secapin.

The use of bee venom (Apis mellifera L., BV) occasionally causes side effects such as inflammation and allergic reactions in the recipients. Several case reports also suggested the treatment of BV has some limitations in its clinical uses, due to the occurrence of dermal necrosis and anaphylatic reactions. It is generally understood that bee venom allergy is mainly the result of its allergic component, phospholipase A2 (PLA2). The present study was aimed to generate PLA2-free bee venom (PBV) and evaluate its efficacy as skin care and cosmetic preparation, comparing with original bee venom (BV). Our results showed that both BV and PBV exhibited significant protective effects in UVB-irradiated human keratinocyte (HaCaT) and human dermal fibroblast (HDF) cells and they also induced type I collagen synthesis in UVB-irradiated HDF cells except BV at 3 μg/ml. Furthermore, BV and PBV showed the inhibition of UVB-stimulated matrix metalloproteinase-1 (MMP-1), a major collagen degrading enzyme in skin. However, BV, unlike PBV, exhibited strong cytotoxicities in skin cells (both HaCaT and HDF) at its working concentrations of anti-wrinkle effect. The underlying cell signaling mechanisms of anti-wrinkle effects of BV and PBV were demonstrated by the activation of ERK1/2, and p38. Conclusively, PBV appears to be the bee venom of choice with less cytotoxicity and higher efficacy on UVB-irradiated skin cells in comparison with original bee venom (BV). Therefore, PBV can better be used as a cosmetic ingredient exhibiting excellent anti-wrinkle effect against photoaging than original BV.

Inhibitor cysteine knot (ICK) peptides exhibit ion channel blocking, insecticidal, and antimicrobial activities, but currently, no functional roles for bee-derived ICK peptides have been identified. In this study, a bee (Apis cerana) ICK peptide (AcICK) that acts as an antifungal peptide and as an insecticidal venom toxin was identified. AcICK contains an ICK fold that is expressed in the epidermis, fat body, or venom gland and is present as a 6.6-kDa peptide in bee venom. Recombinant AcICK peptide (expressed in baculovirus-infected insect cells) bound directly to Beauveria bassiana and Fusarium graminearum, but not to Escherichia coli or Bacillus thuringiensis. Consistent with these findings, AcICK showed antifungal activity, indicating that AcICK acts as an antifungal peptide. Furthermore, AcICK expression is induced in the fat body and epidermis after injection with B. bassiana. These results provide insight into the role of AcICK during the innate immune response following fungal infection. Additionally, we show that AcICK has insecticidal activity. Our results demonstrate a functional role for AcICK in bees: AcICK acts as an antifungal peptide in innate immune reactions in the body and as an insecticidal toxin in venom. The finding that the AcICK peptide functions with different mechanisms of action in the body and in venom highlights the two-pronged strategy that is possible with the bee ICK peptide.

Background: Propionibacterium acnes (P. acnes) is a major contributing factor for the inflammatory reaction of acne. Bee venom (BV) has been traditionally used to the treatment for inflammatory diseases. This study examined the anti-inflammatory effect of BV on P. acnes-induced inflammatory animal model. Methods: P. acnes were intradermally injected into both left and right ear of ICR mice. After injection, different concentrations of BV (1, 10 and 100 μg) mixed with 0.05 g of Vaseline was applied to the surface of the right ears of mice. Results: Histological observation revealed that P. acnes induced a considerable increase in the number of infiltrated inflammatory cells. However, BV treatment showed markedly reduced these reactions. Also, expression levels of TNF-α, and IL-1β were significant reduced in BV treated mice compared with P. acnes injected mice. The binding activity of NF-κB and AP-1 were increased in the P. acnes and Vaseline groups. In contrast, this enhancement of binding activity was markedly withdrawn after treatment with BV. Conclusion: In conclusion, this study indicates that BV has potential as an anti-acne agent and may be useful in the pharmaceutical and cosmetic industries.

Propionibacterium acnes (P. acnes) cause an inflammatory acne that plays an important role in the pathogenesis of acne by inducing inflammatory mediators. Bee venom therapy has been used in oriental medicine for the relief of pain and the treatment of inflammatory diseases. However, a direct effect of bee venom in skin inflammation has not been established. The purpose of this study was to investigate anti-inflammatory properties of bee venom in skin inflammation stimulated by heat-killed P. acnes using human keratinocytes and monocytes cell line. P. acnes stimulates the production of proinflammatory cytokines such as interleukins-1β, -8, interferon-γ and tumor necrosis factor-α in HaCaT and THP-1 cells. Bee venom effectively inhibits the secretion of IL-1β, IL-8, IFN-γ, and TNF-α. P. acnes treatment activates the expression of TLR2, which results in IL-8 expression. However, bee venom treatment reduces the expression of TLR2 and IL-8. Based on these results, bee venom has effects on anti-inflammatory activity against P. acnes in HaCaT and THP-1 cells.

For the investigation of the stability of purified bee venom(PBV) during the treatment in the temperature range of 50℃ to 120℃ for 24 hours, respectively, melittin contents, antibacterial effects, and cell regenerations were investigated. The changes in the melittin contents of PBV were not significantly different by treatment temperature below 70℃ for 24 houes and 80℃ for 4 hours. However the melttin contents is great decline after 24 hours above 80℃ for 24 hours. Antibacterial effects is not change below 80℃ for 4 houes but significantly decrease above 80℃ for 24 hours. Cell regenerations of PBV on human dermal fibroblast decreased at 80℃ for 24 houes, showing a significant difference from the below 80℃ for 4 houes. Through the temperature stability of PBV results of this study, it was treated that the melittin contents, antibacterial effects and cell regeneration effects of PBV could be maintained above 80℃ for 4 hours.

Osteoarthritis is one of the commonest causes associated with age-related damage of articular cartilage. Non-steroidal anti-inflammatory drugs are commonly used in osteoarthritic patient. However, long term administration of these drugs results gastrointestinal disorders. Though, most studies have demonstrated in the past that bee venom has therapeutic effect on diseases related to inflammation and pains, but its anti-inflammatory properties have not been so far studied on inflamed chondrocytes (LPS induced) invitro. For the purpose, the study was carried out to determine the effect of bee venom on porcine articular chondrocyte cell using microarray. In this study, we found that 2,235 significantly associated gene (1,404 up-regulated genes and 831 down-regulated genes) that were expressed on inflamed and non inflamed chondrocytes during proliferation. Among the 1,404 up-regulated genes and 831 down-regulated genes, known genes were 372 and 237, respectively. On the other hand, bee venom significantly reduced expression of fetuin involved in acute inflammatory reaction. Our results suggest that this study could be useful database in gene expression profiling of chondrocyte cell treated with bee venom.

서양종 꿀벌 독의 채집시기와 지역에 따른 봉독의 성분 변화 및 약리효과에 미치는 영향을 검토하였다. 채집시기는 5월부터 9월까지, 채집지역은 전국 35개 지역으로부터 채취한 봉독을 대상으로 2010년과 2011년 2년에 걸쳐 동일 지역에서 동일한 방법으로 봉독을 채취하였다. 채취한 봉독은 액체크로마토그래피를 통해 멜리틴과 아파민 그리고 포스포리파아제 A2의 성분 함량을 분석하였다. 그 결과 채집시기와 지역에 따른 성분에 유의한 차이는 확인되지 않았다(One way-ANOVA, Duncan's test (=0.05)). 봉독의 성분은 채집시기와 지역에 관계없이 멜리틴 , 아파민 그리고 포스포리파아제 A2는 을 차지하였다. 이상의 결과로부터 봉독은 채취시기에 따른 주요 성분은 차이를 갖고 있지 않았으며, 이는 꿀벌의 먹이, 사육온도 등 외부 환경이 봉독 분비에 영향을 주지 않는 것으로 사료되었다.

This study was conducted in order to examine the safety of bee venom as an alternative for antibiotics using male ICR mice. Five-week-old male mice received a single intravenous injection of a dried honey bee venom at the concentration of 0.25 mg/kg (a clinical dose) or 0.5 mg/kg through the tail vein and various pathophysiological analyses were performed after three days. No significant differences in changes of body weight were observed between the saline-treated control group and the experimental groups. In the hematological analysis, none of the parameters were affected by bee venom. In blood biochemistry analysis, none of the markers were affected by administration of bee venom. Similarly, there were no significant effects on markers for liver, kidney, and skeletal muscle functions in all treated- groups. On macroscopic examination, no remarkable lesions were detected in these organs. Because there were no adverse effects of the bee venom in a single intravenous toxicity test for three days, it was concluded that bee venom could be a candidate for a safe natural antibiotic for use in the animal production industry.

Experiments were conducted in order to assess the healing effect of bee venom (BV) cream on full-thickness skin wounds in rabbits. BV cream was compared with silver sulfadiazine (SS) as a topical medicament against a control on experimentally created full-thickness wounds. Two wounds measuring 2 × 2 cm were created bilaterally (four wounds/rabbit) on the dorsolateral aspect of the trunk of seven New Zealand white rabbits. Wound treatments were evenly distributed on four sites, using a Latin square design. The contact layer of wounds was treated with physiological saline (control), SS cream, and BV cream over a period of 28 days. Assessment of wound healing was based on scab hardness, wound exudates, wound area, unepithelialized granulation tissue, and histopathological findings. Topical application of BV and SS creams to wounds resulted in reduced inflammation, debridement of necrotic tissue, and promoted granulation and epithelialization. Wound healing was faster, with statistical significance in BV and SS treatments, compared to the control (P<0.05). Treatment with BV evoked an anti-inflammation effect in a rabbit model. BV cream produced a wound healing effect similar to that of commercially available SS cream. Anti-inflammation effect as a topical treatment with BV cream appears to be better than that with SS cream. These results suggest that topical application of BV cream may be an alternative treatment for full-thickness skin wounds.

We present evidence that the serine protease found in bumblebee (Bombus terrestris) venom exhibits fibrin(ogen)olytic activity. Compared to honeybee (Apis mellifera) venom, bumblebee venom contains a higher content of serine protease, which is one of its major components. Venom serine proteases from bumblebees did not cross-react with antibodies against the honeybee venom serine protease. We provide functional evidence indicating that B. terrestris venom serine protease (Bt-VSP) acts as a fibrin(ogen)olytic enzyme. Bt-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products. However, Bt-VSP is not a plasminogen activator, and its fibrinolytic activity is less than that of plasmin. Taken together, our results define roles for Bt-VSP as a prothrombin activator, a thrombin-like protease, and a plasmin-like protease, providing significant support for thepotential use of bumblebee venom serine protease as a clinical agent.

Bee venom is a rich source of pharmacologically active substances. In this study, we identified a bumblebee (Bombus ignitus) venom Kunitz-type serine protease inhibitor (Bi-KTI) that acts as a plasmin inhibitor. Bi-KTI showed no detectable inhibitory effect on factor Xa, thrombin, or tPA. However, it strongly inhibited plasmin, although this inhibitory ability was two-fold weaker than that of aprotinin. The activities of B. ignitusvenom serine protease (Bi-VSP) and plasmin in the presence of Bi-KTI indicate that Bi-KTI targets plasmin more specifically than Bi-VSP. These findings demonstrate a novel mechanism for bee venom by which Bi-KTI acts as an antifibrinolytic agent, raising interest in Bi-KTI as a potential clinical agent.

Bee venom contains a variety of protein allergens, including serine proteases. Additionally, bee venom has been used in therapeutic application through immunotherapy for bee venom hypersensitivity and venom therapy as an alternative medicine. Here we present a novel view of the application of bee venom through which bee venom serine protease exhibits fibrin(ogen)olytic activity. Compared to honeybee venom, bumblebee venom contains a larger amount of a serine protease as one of its major components. Immunologically, venom serine proteases from bumblebees did not show cross-reactivity with the honeybee venom serine protease. We provide functional evidence indicating that bumblebee (Bombus terrestris) venom serine protease (Bt-VSP) acts as a fibrin(ogen)olytic enzyme. Bt-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products, defining roles for Bt-VSP as a prothrombin activator, a thrombin-like protease, and a plasmin-like protease. However, Bt-VSP did not activate plasminogen and the fibrinolytic activity of Bt-VSP is less than plasmin. These findings offer insight into the allergic reaction sequence of bee venom serine protease and its potential usefulness as a clinical agent in the field of hemostasis and thrombosis.

Bee venom contains a variety of peptides and enzymes, including serine proteases. While the presence of serine proteases in bee venom has been demonstrated, the role of these proteins in bee venom has not been elucidated. Furthermore, there is currently no information available regarding the melanization response or the fibrin(ogen)olytic activity of bee venom serine protease, and the molecular mechanism of its action remains unknown. Here we show that bee venom serine protease (Bi-VSP) is a multifunctional enzyme. In insects, Bi-VSP acts as an arthropod prophenoloxidase (proPO)-activating factor (PPAF), thereby triggering the phenoloxidase (PO) cascade. Bi-VSP injected through the stinger induces a lethal melanization response in target insects by modulating the innate immune response. In mammals, Bi-VSP acts similarly to snake venom serine protease, which exhibits fibrin(ogen)olytic activity. Bi-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products, defining roles forBi-VSP as a prothrombin activator, a thrombin-like protease, and a plasmin-like protease. These findings provide a novel view of the mechanism of bee venom in which the bee venom serine protease kills target insects via a melanization strategy and exhibits fibrin(ogen)olytic activity.