• Journal of Acupuncture Research
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• 제19권2호
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• pp.92-104
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• 2002

Objective : Bee venom (BV) has traditionally been used in Oriental medicine to relieve pain and to treat inflammatory disease such as rheumatoid arthritis (RA). Autoimmunity to type II collagen (CII) may involve in the pathogenesis of RA. This study was performed to evaluate the effect of BV on type II collagen induced arthritis (CIA) with the naked eye, a immunohistochemical method and the examination of histology. Method : Male mice were immunized by subcutaneously injection of an $200{\mu}g$ emulsion mixed with bovine CII and complete Freund's adjuvant (CFA) twice for two weeks. In the control group, normal saline was injected, and in the experimental group, BV was applied. Result : The incidence of arthritis, the mean arthritis index and the number of the arthritic limbs of the BV group were all significantly lower than those of the control group. Among the pro-inflammatory cytokines, the production of $TNF-{\alpha}$ in the BV group was also suppressed compared with the control group, but $IL-1{\beta}$ was not. The examination on the histopathology of joints of CIA mice showed the effect of Bee Venom Herbal Acupuncture on the arthritis. Conculusion : Treatment with BV resulted in inhibition of development of arthritis and immune responses to CII.

• Journal of Applied Biological Chemistry
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• 제62권4호
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• pp.347-353
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• 2019

Stings of jellyfish, which frequently occur in a warm season, cause severe pain, inflammation and sometimes irreversible results such as the death. Harmful venoms from jellyfish, therefore, have been studied for finding the therapeutic agents to relieve pain or to neutralize toxic components. However, it is still unclear if and how jellyfish venom reveal neuronal toxicity even though pain induction seems to result from the activation of nociceptors such as nerve endings. In this study, using HT-22 cell line, we investigated neurotoxic effects of the venom of Chrysaora pacifica (CpV) which appears in South-East ocean of Korea. In 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, CpV significantly reduced the viability of HT-22 cells in a dose-dependent manner. Additionally, in 2',7'-Dichlorofluorescin diacetate fluorescence test under the culture condition lacking dominant inflammatory factors, CpV remarkably increased the production of intracellular reactive oxygen species (ROS). Reduced responsive fluorescence to Rhodamine123 and increased expression of intracellular cytochrome c were also observed in HT-22 cells treated with CpV. These indicate that CpV-reduced viability of HT-22 cells may be due to the activation of apoptotic signalings mediated with oxidative stress and mitochondrial dysfunction. Furthermore, removing Ca²⁺ ion or adding N-acetyl-Lcystein remarkably blocked the CpV effect to reduce the viability of HT-22 cells. The findings in this study clearly demonstrate that CpV may activate Ca²⁺-mediated ROS signalings and mitochondrial dysfunction resulting in neuronal damage or death, and suggest that blocking Ca²⁺ pathway is a therapeutic approach to possibly block toxic effects of jellyfish venoms.

• 한국미생물·생명공학회지
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• 제15권2호
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• pp.135-139
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• 1987

Aspergillus 속 균주 MK-24로부터 venom proteinase inhibitor의 생산조건을 검토한 결과 질소원으로서는 유기능질소가 균생육에 있어서는 매우 좋았으나 물질생성에 있어서는 무기능질소보다 못하였다. 무기질소원으로서는 sodium nitrate가 가장 좋은 효과를 보였다. 단소원으로서는 glucose가 대부분 당류와 비슷한 결과를 나타내었고 특히 arabitol, xylitol, salicin 등은 본 균주가 단소원으로 사용치 못하는 것으로 추정되었으며, vitamin 류는 물질생성에 무관하였으며 금속염류로서는 효과적인 것은 없으나, Ag$^+$, Co$^{++}$, $Zn^{++}$ 등은 억제하였다. 배양온도와 pH는 각각 3$0^{\circ}C$와 pH 5가 균의 생육과 저해물질생성에 제일 적당하였고, 단소원과 질소원으로서 glucose 2%, NaNo$_3$ 0.3%을 가한 액체배지에서 7일간 정치배양 하였을 때 저해물질생육이 가장 좋았다.

• Journal of Animal Science and Technology
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• 제48권6호
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• pp.933-942
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• 2006

본 실험은 생봉독 처리가 돼지의 체액성 면역반응에 대한 생봉독의 효과를 조사하기 위하여 실시하였다. 혈중 Immunoglobulin(Ig) G, IgA, IgM의 농도에 미치는 생봉독의 효과를 조사하기 위하여 모돈 3두에서 생산한 자돈 20두(LY ×D)를 생봉독 처리군과 대조군으로 각각 10두씩 배치하였다. 생봉독 처리군은 출생시와 3일령에 교소혈 (GV-1), 해문혈(ST-25) 및 두구혈(CV-8), 6일령에 거세, 단미 창상부위에 생봉 1마리씩 직침 시술하였고, 대조군은 생리식염수 1ml를 동일한 혈위에 주입하였다. 혈중 Ig 농도 측정을 위하여 출생시와 3일, 7일, 14일 및 21일령에 채혈하여 Immunoturbidimetric method로 IgG, IgM, IgA를 측정하였다. 지표항원으로 사용한 돈 콜레라와 위축성비염 백신에 대한 생봉독의 항체 생성효과를 조사하기 위하여 모돈 5두에서 생산된 자돈 40두(LY×D)를 생봉독 처리군과 대조군으로 각각 20두씩 공시하였다. 생봉독 처리군은 출생시에 교소혈(Jiao-Chao, GV-1), 해문혈(Hai-Men, ST-25) 및 두구혈(Du-Kou, CV-8)에, 3일령에 단미 및 거세시술 시 창상부위에 생봉독을 처리하였고, 21일령 이유시에는 교소혈(GV-1)과 백회혈(Bai- Hui, GV-20)에 생봉독을 처리하였다. 지표항원으로 위축성비염 백신은 24일령과 44일령에 접종하였고 돈콜레라 백신은 44일령과 64일령에 각각 접종하였다. 항체가 분석용 혈액은 24, 34, 44, 54 및 74일령에 채혈하여 위축성비염은 시험관응집반응, 돈콜레라는 ELISA법에 의하여 항체가를 조사하였다. IgG 농도는 처리군에서 출생시 339.52, 3일령에 366.48, 7일령에 296.52, 14일령에 242.06, 21일령에는 219.06mg/dl이었고 대조군은 각각 347.10, 333.14, 243.28, 205.18 및 191.58mg/dl 이었다. 처리전(출생시)에는 처리군과 대조군간에 유의차가 없었으나 처리군의 IgG 농도가 대조군에 비하여 3일령 10.3% (P<0.02), 7일령에는 21.9% (P<0.01), 14일령에는 18.0% (P<0.07), 21일령에는 14.3(P<0.07) 더 높게 나타났다. IgA와 IgM의 농도는 전체기간동안 처리군과 대조군간에 유의차가 인정되지 않았다. 돈콜레라 virus에 대한 항체역가는 처리군에서 대조군에 비하여 24일령 때 57.0%(P<0.03), 34일령 때 74.6% (P<0.006), 44일령에 48.6% (P<0.017), 54일령 때 45.0% (P<0.16), 74일령 때 44.4%(P<0.006)가 유의하게 높은 것으로 나타났다. 위축성비염 원인균인 Bordetella bronchiceptica에 대한 항체역가는 처리군이 대조군에 비하여 34일령 때는 39.7%(P<0.002), 44일령 때 31.9% (P<0.02), 54일령 때에 33.4%(P<0.01), 74일령 때에는 57.3%(P<0.007)가 높게 나타나 접종일인 24일령을 제외하고 전체기간에 걸쳐 높은 항체수준을 보였다. 위의 결과를 종합해 볼 때 돼지에 대한 생봉독 처리는 면역기능향상 효과가 있는 것으로 사료된다. 이와 같은 결과를 볼 때 돼지에 대한 생봉독의 처리는 양돈 생산성면에서 생봉독 처리는 돼지의 생존율을 높이고, 증체량이 높았다는 조 등(2005)의 보고를 뒷받침하고 있어 양돈 산업발전에 기여할 것으로 사료된다.

• The Korean Journal of Physiology and Pharmacology
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• 제9권3호
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• pp.179-186
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• 2005

Whole bee venom (WBV) and its major component, melittin, have been reported to induce long-lasting spontaneous flinchings and hyperalgesia. The current study was designed to elucidate the peripheral and spinal mechanisms of N-methyl-D-aspartate (NMDA) and non-NMDA receptors by which intraplantar (i.pl.) injection of WBV and melittin induced nociceptive responses. Changes in mechanical threshold and flinching behaviors were measured after the injection of WBV (0.04 mg or 0.1 mg/paw) and melittin (0.02 mg or 0.05 mg/paw) into the mid-plantar area of a rat hindpaw. MK-801 and CNQX (6-cyano-7-nitroquinoxaline-2,3-dione disodium) were administered intrathecally (i.t. $10{\mu}g$) or i.pl.($15{\mu}g$) 15 min before or i.t. 60 min after i.pl. WBV and melittin injection. Intrathecal pre- and postadministration of MK-801 and CNQX significantly attenuated the ability of high dose WBV and melittin to reduce paw withdrawal threshold (PWT). In the rat injected with low dose, but not high dose, of WBV and melittin, i.pl. injection of MK-801 effectively suppressed the decrease of PWTs only at the later time-points, but the inhibitory effect of CNQX (i.pl.) was significant at all time-point after the injection of low dose melittin. High dose WBV- and melittin-induced spontaneous flinchings were significantly suppressed by i.t. administration of MK-801 and CNQX, and low dose WBV- and melittin-induced flinchings were significantly reduced only by intraplantarly administered CNQX, but not by MK-801. These experimental flinchings suggest that spinal, and partial peripheral mechanisms of NMDA and non-NMDA receptors are involved in the development and maintenance of WBV- and melittin-induced nociceptive responses.

• Journal of Acupuncture Research
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• 제22권2호
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• pp.123-132
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• 2005

Melittin,cationic 26-amino acid, is the principal component of the bee venom (BV) which has been used for treatment of inflammatory disease such as arthritis rheumatism NF-kB is activated by subsequent release of inhibitory IkB via activation of a multisubunit IkB kinase (IKK). We previously found that melittin bind to the sulfhydryl group of p50, a subunit of NF-kB. Since sulfhydryl group is present in kinase domain of IKKa and IKKb, melittin could modify IKK activity by protein-protein interaction. We therefore examined effect of melittin on IKK activities in sodium nitroprusside (SNP)-stimulated synoviocyte obtained from RA patients. Melittin suppressed the SNP-induced release of IkB resulted in inhibition of DNA binding activity of NF-kB and NF-kB-dependent luciferase activity. Consistent with the inhibitory effect on NF-kB activation, IKKa and IKKb activities were also suppressed by melittin. Surface plasmon resonance analysis realized that melitin binds to IKKa $(Kd\;=\;1.34{\times}10-9M)$ and IKKb$(Kd\;=\;1.0{\times}10-9M)$. Inhibition of IKKa and IKKb resulted in reduction of the SNP-induced production of inflammatory mediators NO and PGE2 generation. The inhibitory effect of melittin on the IKKs activities, binding affinity of melittin to IKKs, and NO and PGE2 generation were blocked by addition of reducing agents dithiothreitol and glutathione. In addition, melittin did not show inhibitory effect in the transfected Synoviocytes with plasmid carrying dominant negative mutant IKKa (C178A) and IKKb (C179A). These results demonstrate that melittin directly binds to sulfhydryl group of IKKs resulting in IkBrelease, thereby inhibits activation of NF-kB and expression of genes involving in the inflammatory responses.

• Journal of Acupuncture Research
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• 제40권4호
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• pp.368-376
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• 2023

Background: Although bee venom (BV) has clinical benefits in osteoarthritis and rheumatoid arthritis, it has not been tested as treatment for gouty arthritis. Moreover, in vitro, BV has been proven to exhibit anti-inflammatory and positive effects on osteoarthritis, but only limited evidence can confirm its beneficial effects on gout. Thus, this study aims to assess the anti-inflammatory effects of BV on monosodium urate (MSU)-induced THP-1 monocytes. Methods: THP-1 monocytes were differentiated into mature macrophages using phorbol 12-myristate 13-acetate (PMA) and pretreated for 6 hours with BV and a Caspase-1 inhibitor in a physiologically achievable range of concentrations (BV, 0.1-1 ㎍/mL; Caspase-1 inhibitor, 1-10 μM), followed by MSU crystal stimulation for 24 hours. The secretions of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), IL-6, IL-8, cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2), and nitric oxide (NO) were increased in the MSU crystal-stimulated THP-1 cells. Results: Caspase-1 inhibitors suppressed the production of all mediators in a dose-dependent manner. BV worked on equal terms with Caspase-1 inhibitors and showed more satisfactory effects on TNF-α, PGE2, COX-2, and inducible nitric oxide synthase (iNOS). Moreover, the western blot analysis revealed that BV regulated the transcriptional levels of these mediators via the suppression of extracellular signal-regulated kinase (ERK) pathway activation. Conclusion: The results of the present study clearly suggest that BV inhibits MSU-induced inflammation in vitro, suggesting a possible role for BV in gout treatment.

• The Korean Journal of Physiology and Pharmacology
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• 제10권6호
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• pp.297-302
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• 2006

Melittin, a major component of bee venom, produces a sustained decrease in mechanical threshold, and an increase in spontaneous flinchings and paw thickness, which are characteristics similar to those induced by whole bee venom. Melittin-induced nociception has been known to be modulated by the changes in the activity of excitatory amino acid receptors, voltage-dependent calcium channels, cyclooxygenase and serotonin receptors. The present study was undertaken to investigate the role of calcium chelators (TMB-8 & Quin 2) in melittin-induced nociceptive responses. Changes of mechanical threshold and spontaneous flinching behaviors were measured at a given time point following intraplantar injection of melittin ($30{\mu}g/paw$). Intrathecal or intraplantar pre-administration and intrathecal posttreatment of TMB-8 and Quin 2 significantly prevented the melittin-induced reduction of mechanical threshold, and intraplantar or intrathecal pre-treatment of TMB-8 and Quin 2 suppressed melittininduced flinching behaviors. These results indicate that calcium ion in the spinal dorsal horn neurons and peripheral nerves plays an important role in the production and maintenance of mechanical allodynia and spontaneous pain by melittin.

• Journal of Acupuncture Research
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• 제23권2호
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• pp.33-46
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• 2006

Alzheimer's disease (AD) is the most prevalent form of neurodegenerative disease associated with aging in the human population. This disease is characterized by the extracellular deposition of beta-amyloid peptide $(A{\beta})$ in cerebral plaques. $A{\beta}$ is derived from the ${\beta}-amyloid$ precursor protein (APP) by the enzymes, ${\beta}-$ and ${\eta}o-secretase$. Compounds that ${\beta}-$ or ${\eta}o-secretase$ inhibit activity, can reduce the production of $A{\beta}$ peptides, and thus have therapeutic potential in the treatment of AD. Increasing body of evidence has been demonstrated that Bee Venom(BV) Acupuncture could compete with complex protein involving in multiple step of $NF-{\kappa}B$ activation and exert the anti-inflammatory potential of combined inhibition of the prostanoid and nitric oxide synthesis systems by inhibition of IKK and $NF-{\kappa}B$. In this study, I investigated possible effects of BV on memory dysfunction caused by lipopolysaccharide (LPS) and $A{\beta}$ through inhibition of secretases activities and $A{\beta}$ aggregation. I examined the improving effect of BV on the LPS (2.5 mg/Kg, i.p.)-induced memory dysfunction using passive avoidance response and water maze tests in the mice. BV (0.84, $1.67\;{\mu}g/ml$) reversed the LPS-induced memorial dysfunction in dose dependent manner. BV also dose-dependently attenuated LPS-induced ${\beta}$ and ${\eta}o-secretase$ activities in cerebral cortex and hippocampus of the mice brain. This study therefore suggests that BV acupuncture method may be useful for prevention of development or progression of AD.

• International Journal of Industrial Entomology and Biomaterials
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• 제35권1호
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• pp.39-44
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• 2017

The venome of Phoneutria nigriventer spider has been shown to have side effects including severe painful erections that last for hours. PnTx2-6, a toxin from P. nigriventer spider venom, modulates voltage gated $Na^+$ channels and activation of nitric oxide (NO) production. NO is essential for the regulation of blood flow and pressure. Therefore, PnTx2-6 is expected to be effective not only for erectile dysfunction but also for cardiovascular diseases. A previously has reported cDNA clone for PnTx2-6 toxin, which was expressed in E. coli cytoplasm. We created the same clone and expressed it in a bacterial expression system. PnTx2-6 increased the genes expression of superoxide dismutase 1, glutathione peroxidase 1, and sulfiredoxin 1. We hypothesized that recombinant PnTx2-6 may indirectly regulate blood flow and pressure, resulting in NO production in human umbilical vein endothelial cells (HUVEC). These data suggest differential regulation of the vascular ageing process, which may contribute to the anatomic heterogeneity of atherosclerosis. The results of this study may be used for the emergency treatment of sudden cardiovascular disease caused by ageing.