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Effects of Sweet Bee Venom on the Central Nervous System in Rats -using the Functional Observational Battery-

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(1)ISSN 2093-6966. Journal of Pharmacopuncture. 19 http://dx.doi.org/10.3831/KPI.2011.14.3.019. Original Article. Sweet BV 시술이 Rat의 중추신경계에 미치는 영향 - 기능관찰 종합평가를 이용하여-. Received : May 31, 2011 Accepted : Jun 3, 2011. Effects of Sweet Bee Venom on the Central Nervous System in Rats -using the Functional Observational BatteryJoong Chul An 1, Ki Rok Kwon2*. KEY WORDS: Melittin; Sweet Bee Venom; functional observational battery (FOB); central nervous system. 1. 2. Sangji University Graduate School of Oriental Medicine, Wonju, Korea Research Center of Pharmacopuncture Medicine, Korean Pharmacopuncture Institute, Seoul, Korea. ABSTRACT 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.. cc This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original work is properly cited.. 2011 Korean Pharmacopuncture Instltute. Corresponding Author Ki Rok Kwon. Research Center of Pharmacopuncture Medicine, Korean Pharmacopuncture Institute, Seoul, Korea Tel: +82-2-2658-9051 E-mail: [email protected] http://pharmacopuncture.co.kr.

(2) 20. Journal of Pharmacopuncture 제 14권 제 3호( 2011년 9월). I. 서론. Sweet BV. 2. Apis melifera. 2.1 Sprague-Dawley SD rat. 106.7. 139.1g. 5. rat 1 anaphylactic shock Bee Venom Sweet BV 40-50 melittin. 10-0744755. melittin. Sweet BV. Sweet Sweet BV 2840 gel filtration. rat. 1. Appendix 2 20.3 23.6 40.5 58.5 Teklad Certified Irradiated Global 18 Protein Rodent Diet 2918C 2005-79 Good Laboratory Practice GLP. 2.2 36. Sweet BV. 32 8. rat battery. Sweet BV Functional observational. FOB. II. 재료 및 방법. 2.3 1 Sweet BV 60. 4. 0.1 0.007 100 0.7 0.175 0.35 Sweet BV normal saline. 1 Sweet BV 0.2 LA230S, CP323S, Sartorius, Germany Normal Saline vortex mixer 0.875, 1.75 3.5. 2. 4. 0.4. 1.

(3) 21. 3. 3.1 1 1. 1. data type: quantitative value way analysis of variance ANOVA p 0.05. 1. 2. Dunnett's tp 0.05 p 0.01 data type: rank Kruskal-Wallis p 0.05 Steel s. test. 1. test. 3.2. One-. 1 test. III. 결과 4. 1 4.1 1. 1. Sweet BV 0. 0.5, 1, 2. 4 Table 1-2. 4.2 Functional observational battery, FOB 1 2 2 Methods and criteria for evaluation of functional observational battery Attachment home cage open field hand held. 50 10 dB. 2. Home cage 0.175, 0.35 0.7 Sweet BV body posture behavior ease of removal from cage 0 0.5, 1, 2 4 Table 3-1, Appen -dix 3-1. 30 white noise 3. Open field parameters 0.175 2 counts. Sweet BV p 0.05 0.5. rearing p 0.01. 5. number of urine pools parameter SAS Ver. 9.1.3, SAS Institute Inc., U.S.A.. p 0.01. 0.

(4) Journal of Pharmacopuncture 제 14권 제 3호( 2011년 9월). 22. respiration rate stereotypy unusual behavior tremor convulsion gait arousal level number of fecal pellets 0.35 2 2. Sweet BV p 0.01 p 0.05. aerial righting reflex hindlimb landing foot splay forelimb grip strength hindlimb grip strength 0 0.5, 1, 2 4 Table 3-4, Appendix 3-4. 0.5 6. 0.175 0.35 rectal temperature 1, 2 4. 0.7 0.5. Sweet BV p 0.01. 0.7. Sweet BV 0. 0.5,. Sweet BV p 0.05. 1. Table 3-5, Appendix 3-5 Table 3-2, Appendix 3-2. IV. 고찰 4. Hand held 0.175, 0.35. 0.7. Sweet BV fur and skin appearance piloerection soiled lower abdominal region lacrimation salivation palpebral closure pupil size pupillary reflex muscle tone extensor-thrust reflex 0 0.5, 1, 2 4 Table 3-3, Appendix 3-3. 4. 3. testing protocol guideline 5. 0.175, 0.35 0.7 visual response click response. Sweet BV touch response tail pinch response. GLP.

(5) 23. 4. Functional observational battery, FOB FOB. Fig 2. The practical aspect of functional observational battery(FOB) The FOBs were conducted by trained observers working together in pairs; apart from two persons being required for several of the tests within the FOB, this also enabled a consensus view of any equivocal observations.. 30 ICHS7A FOB. home cage home cage. FOB Apis melifera. rat. center circle open field activity, sensorimotor reflex test/physiological measurements Fig. 1-2. 40. onidase. phospholipase A2 PLA2 histamine. hyalurmelittin. Fig 1. The process of functional observational battery(FOB) The sequence of observations in the FOB is consist of five parameter. The parameter is next follows; home cage observations, home cage removal and handling, open field activity, sensorimotor reflex test/physiological measurements and observations throughout FOB.. Sweet BV. B. NF- B. melittin nuclear factorlipopolysaccharide.

(6) Journal of Pharmacopuncture 제 14권 제 3호( 2011년 9월). 24. LPS tumor necrosis factor- TNFum nitroprusside SNP melittin 4. sodi-. 0. 0.7. Sweet BV 0.07. melittin. 0.1. 0.4. 0.007 Sweet BV melittin 50-100. Sweet BV human 60 SD 0.175, 0.35 0.7. FOB. Sweet BV Sweet BV 0.175. FOB 0.175 BV. Sweet. 2. V. 결론. 0.5 0 parameters 0.35 Sweet BV 2 0.5. FOB rat. Sweet BV. 2 parameters 0.7 Sweet BV 0.5 parameters 0.175, 0.35 0.7 0.5. BV. Sweet BV. 1. Home cage Sweet 2. Open field. Sweet BV. Sweet BV. 3. Hand held. 4. Sweet BV rat. Sweet BV. 50. 5.. Sweet BV. 100 0.175. Sweet BV. Sweet BV.

(7) 25. VI. References 1. Academic committee of KPI. Pharmacopuncturology. Elsevier Korea. 2008;167-199. 2. Kwon KR, Koh HK. An Experimental Study with Bee Venom Therapy on Anti-inflammatory and Analgesic Effects. The Journal of Korean Acupuncture & Moxibustion Society. 1998;15(2): 97-103. 3. Ko HK. Experimental Studies on the Effect of Bee Venom Theraphy on the Analgesic, Antipyretic and Anti-inflammatory Action. Korean Journal of Oriental Medicine. 1992;13(1):283-292. 4. Kwon KR, Koh HK, Kim YS, Park YB, Kim CH, Kang SK, The Effect of Bee Venom Aqua -acupuncture on the Antitumor and Immune Response in the Epithelioma by 3 - MCA. The Journal of Korean Acupuncture & Moxibustion Society. 1997;14(2):151-172. 5. Lee SN, Hong SY, Jo HC, Byeon IJ, Song HS,. Kim GH, The Clinical Study on Bee Venom Acupuncture Treatment on Osteoarthritis of Knee Joint. The Journal of Korean Acupuncture & Moxibustion Society. 2003;20(5):73-8. 6. Wang WH, Ahn KB, Lim JK, Jang HS, Clincal Investigation Compared with the Effects of the Bee Venom - Acupuncture on Knee Joint with Osteoarthritis. The Journal of Korean Acupuncture & Moxibustion Society. 2001;18(3):35-47. 7. Kim TH. Kang KS, Kwon KR. Three cases of affections of the hip treated with Korean BeeVenom therapy. Journal of Korean Pharmacopuncture Institute. 2001;4(3):122-130. 8. Lee SH, Lee HJ, Baek YH, Kim SY, Park JK, Hong SJ, Yang HI, Kim KS, Lee JD, Choi DY, Lee DI. Effects of Bee Venom on the pain,. edema, and acute inflammatory reactant of Rheumatoid Arthritis patients. The Journal of Korean Acupuncture & Moxibustion Society. 2003;20(2):77-84. 9. Lee SH, Hong SJ, Kim SY, Yang HI, Lee JD, Choi DY, Lee DI, Lee YH. Randomized Controlled Double Blind Study of Bee Venom Therapy on Rheumatoid Arthritis. The Journal of Korean Acupuncture & Moxibustion Society. 2003;20(6):80-88. 10. Kim KU, Seo BM, Yun JS, Lee YK, Choi SH, Lee KM, Lim SC, Seo JC, Jung TY, Han SW. The Comparison of Bee Venom Herbal-acupuncture Therapy between Neighboring Acupuncture Points and Neighboring-Remote Acupuncture Points on the Treatment of Lumbar Spine Herniation of Nucleus Pulpous. The Journal of Korean Acupuncture & Moxibustion Society. 2005;22(6):181-187. 11. Lee KM, Lee KS, Yem SC, Jang JH, Yun JY, Hwang BC, Kug YS, Jang JY, Choi JS, Kim YJ, Park JU. A Clinical study of Bee-venom acupuncture treatment on protrusion disc Patients. The Journal of Korean Acupuncture & Moxibustion Society. 2004;21(5):13-25. 12. Jun HJ, Hwang U, Kim JS, Nam SS, Kim YS. Clinical Evaluation of Herniation of nucleus purposus patients treated by Bee venom therapy. The Journal of Korean Acupuncture & Moxibustion Society. 2003;20(5):63-72. 13. Lee DY, Lee KM, Yeom SC, Kim DH, Kim DJ. A Clinical Study of Bee Venom Acupuncture Therapy on Shoulder Pain Patients in Stroke Sequelae. The Journal of Korean Acupuncture & Moxibustion Society. 2006;23(4):69-80. 14. Jeoung KS, Kim SH, Park SK, Lim HJ, Yoon HS, Ahn HJ. Clinical Study on the Effect of Bee Venom Acupuncture Therapy on the Poststroke Pain. The Journal of Korean Acupuncture & Moxibustion Society. 2005;22(3):69-75. 15. Choi YC, Kwon KR, Choi SH. Purification of.

(8) 26. Journal of Pharmacopuncture 제 14권 제 3호( 2011년 9월). Peptide Components including Melittin from Bee Venom using gel filtration chromatography and propionic acid/urea polyacrylamide gel electrophoresis. Journal of Pharmacopuncture. 2006;9(2):105-112. 16. Barbara & Rudolf. Chemistry and Pharmacology of Honey Bee venom. Academic Press. 1986;329-402. 17. Park HJ, Lee SH, Son DJ, Oh KW, Kim KH, Song HS, et al. Antiarthritic effect of Bee Venom. Arthritis & Rheumatism. 2004;50(11):3504-3515. 18. Park HJ, Lee HJ, Choi MS, Son DJ, Song HS, Song MJ, et al. JNK pathway is involved on the inhibition of inflammatory target gene expression and NF- B activation by melittin. J. Inflammation. 2008;5(7):1-13. 19. Kim YJ, Lim CS, Kwon KR. Study of single dose test of Sweet Bee Venom in rats. Journal of Pharmacopuncture. 2009;12(4):5-32. 20. 200560 21. Irwin, S. Comprehensive behavioural assessment; A systematic quantitative procedure for assessing the behavioural and physiologic state of the mouse. Psychopharmacologia: 1968;13:222-257. 22. 1993;9-10, 185-186. 23. 200856 24. Redfern W.S., Wakefield I.D., Prior H., Pollard C.E., Hammond T.G., & Valentin J.P. Safety pharmacology-a progressive approach. Fundamental and Clinical Pharmacology. 2002;16:161-173. 25. Moser V.C., McCormick J.P., Creason J.P., and MacPhail R.C. Comparison of chlordimeform and carbaryl using a functional observational battery. Fundam. Appl. Toxicol. 1988;11:189-206.. 26. Anon. ICHS7A-Note for Guidance on Safety Pharmacology Studies for Human Pharmaceuticals. European Agency for the Evaluation of Medicinal Products, Committee for Proprietary Medicinal Products(CPMP), London, UK. Reference CPMP/ICH/539/00. 2000. 27. Hwang YJ, Lee BC. Clinical Study of Anaphylaxis on Bee-Venom Acupuncture. The Journal of Korean Acupuncture & Moxibustion Society. 2000;17(4):149-159. 28. Youn HM. The Clinical Observation of Anaphylaxis on Bee-Venom Acupuncture. The Journal of Korean Acupuncture & Moxibustion Society. 2005;22(4):179-188. 29. Kwon KR, Chu CS, Park HS, Kim MK, Cha BC, Lee E. Experimental studies of quantitative evalution using HPLC and safety of Sweet Bee Venom. Journal of Pharmacopuncture. 2007;10(2):81-86. 30. Kwon KR, Choi SH, Cha BC. Component Analysis of Sweet BV and Clinical Trial on Antibody Titer and Allergic Reactions. Journal of Pharmacopuncture. 2006;9(2):79-86. 31. Lee JS, Lee JY, Kwon KR, Lee HC. A Study on Allergic response between Bee Venom and Sweet Bee venom Pharmacopunture. Journal of Pharmacopuncture. 2006;9(3):61-77. 32. Renata MS, Jorge A, Hugo V. Structural and functional behavior of biologically active monomeric melittin. J. Molecular Graphics and Modelling. 2007;25:767-772..

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Fig 1. The process of functional observational battery(FOB) The sequence of observations in the FOB is consist of five parameter

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