Journal of Pharmacopuncture (대한약침학회지)

KOREAN PHARMACOPUNCTURE INSTITUTE (대한약침학회)
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The Analgesic Effects of Apitoxin and its Mechanism via JOR and Measuring Expression of mRNA in Phospholipase and TPH using RT-PCR

Jaw Opening Reflex 및 RT-PCR을 이용한 봉독의 진통효과

  • Cho, Kwang-Ho (Dept. of Acupuncture & Moxibustion Oriental Medical College, KyungHee University) ;
  • Lee, Jae-Dong (Dept. of Acupuncture & Moxibustion Oriental Medical College, KyungHee University) ;
  • Park, Dong-Suk (Dept. of Acupuncture & Moxibustion Oriental Medical College, KyungHee University) ;
  • Ahn, Byoung-Choul (Dept. of Acupuncture & Moxibustion Oriental Medical College, KyungHee University)
  • Published : 2000.07.30
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Abstract

The purpose of this study is to prove the analgesic effects of apitoxin and its mechanism via jaw-opening reflex(JOR) and measuring expression of mRNA in Phospholipase and Tryptophan hydroxylase(TPH) using RT-PCR. The experiments were carried out on Sprague-Dawley rats(300-400g) and mastocytoma(P-185 HTR) for JOR and RT-PCR, respectively. Rats anesthetized with thiopental sodium (80mg/kg) were used in the Tooth Pulp stimulation induced JOR. The amplitude of a digastric electromyogram (dEMG) was recorded during the stimulation at an intensity of 1.5 times the threshold for JOR. Apitoxin used in this experiment was diluted with normal saline by 1:1000. Apitoxin was injected intravenously into the test group while normal saline to the control group. However, it was injected directly into the cell of mastocytoma. We referred to base sequence registered in Genbank in designing primers for RT-PCR. The results were as follows; (1)Compared with control group, analgesic effect started to show right after Sprague-Dawely rats were treated with apitoxin($71.50{\pm}8.08$) and lasted for 50 minutes. (2)As a result of the experiment of RT-PCR, we witnessed significant changes in the degree of expression of phospholipase or rate-limiting enzyme of biosynthesis of prostaglandins with $10{\mu}g/ml$ apitoxin.($31.74{\pm}18.98%$, P<0.05) (3)As a result of the experiment of RT-PCR, we witnessed significant changes in the degree of expression of TPH or rate-limiting enzyme in biosynthesis of serotonin with $10{\mu}g/ml$ apitoxin.($131.37{\pm}16.87%$, P<0.05). These results suggest that $10{\mu}g/ml$ apitoxin have the most analgesic effects. This study showed that apitoxin has analgesic effects and held good for 50 minutes. The injection of apitoxin has brought out changes in the degree of expression of phospholipase and TPH. These results strongly suggest that analgesic mechanism by apitoxin is closely related to prostaglandins and serotonin.

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References

  1. 분자의학연구기법 양인명;최영길
  2. 생리학 30명의 의과대학교수(편)
  3. 임상약리학 유지수;황애란
  4. 약리학강의 이우주
  5. 임상약리학 49명의 의과대학교수(편)
  6. 경희의학 v.8 no.3 上丘刺戟이 흰쥐의 開口反射에 미치는 영향 尹相協;曺眞榮;閔炳一
  7. 경희한의대 석사학위논문 捻轉法이 병행된 침자극 및 전침자극이 진통효과에 미치는 영향 김지훈
  8. 경희의과대학 석사논문 RT-PCR을 이용한 흰쥐 송과선 trytophan hydroxylasse유전자 발현의 정량적 분석 박순희
  9. 대한침구학회지 침구학에서의 진통 .소염에 대한 소고 손관영;박영배;강성길
  10. 경희한의대논문집 v.7 金絲注入 자극이 抗炎鎭痛 및 항피로작용에 미치는 효능에 관한 실험적 연구 고형균;최용태
  11. 경희한의대논문집 v.7 當歸水針刺戟이 鎭痛效果에 미치는 영향에 대한 연구 朴快煥
  12. 경희한의대 석사학위논문 附子藥鍼이 鎭痛 및 消炎作用에 미치는 영향에 대한 연구 鄭善喜
  13. 대한한의학회지 v.13 no.1 봉침독요법이 항염, 진통 및 해열에 미치는 효능에 관한 연구 고형균
  14. 경희한의대논문집 v.15 no.1 중완 및 족삼리의 蜂毒요법이 진통효과에 미치는 영향 이종석
  15. 대한한의학회지 v.20 no.1 흰쥐에서 합곡혈 蜂毒약침자극에 의한 개구반사의 반응 김이화;노식;이재동;민병일
  16. 대한침구학회지 v.11 no.1 蜂針에 대한 고찰 권기록;고형균;김창환
  17. 대한침구학회지 v.15 no.1 蜂毒요법에 대한 한의학 최초의 문헌기록: 마왕퇴의서의 蜂毒요법 2례 인창식;고형균
  18. 대한침구학회지 v.14 no.2 蜂毒요법의 최신 연구동향에 대한 고찰 김지영;고형균;김용석(외)
  19. 대한침구학회지 v.16 no.2 Extrusion type의 요추 추간판탈출증 환자의 蜂毒요법을 병행한 한의학적 치료의 임상보고 이병철
  20. 대한침구학회지 v.15 no.1 蜂毒약침요법의 항염증작용에 관한 실험적 연구 김지영;고형균;김용석(외)
  21. 대한침구학회지 蜂毒요법이 항염, 진통작용에 미치는 효능에 관한 실험적 연구 권기록;고형균
  22. 경희한의대 박사학위논문 침, 구 및 홍화수침자극이 흰쥐 뇌부위별 Serotonin 및 Catecholamine함량에 미치는 영향 김영진
  23. 경희대학교 박사학위논문 침, 전침, 자락 및 유향수침자극의 진통작용 및 흰쥐 뇌부위별 serotonin 및 Catecholamine함량에 미치는 영향 김정곤
  24. 경희대학교 석사학위논문 침자가 동통유발된 흰쥐의 TFL, Glucose 및 내분비대사에 미치는 영향 김이화
  25. 경희대학교 박사학위논문 고빈도전침자극의 파형 및 자극시간에 따른 중추신경계 신경세포 및 뇌간의 Serotonin성 신경세포의 활성변화에 미치는 영향 손성세
  26. Textbook of Medical Physiology v.8 Guyton, Arthur C.
  27. Brain Res Brain Res Protoc v.2 no.4 Measurement of trytophan hydroxylase mRNA levels by competitive RT-PCR Clark, M.S.;Russo, A.F. https://doi.org/10.1016/S1385-299X(98)00004-X
  28. Physiol. Rev. v.72 Structure and function of the brain serotonin system Jacobs, B.L.;Azmitia, E.C. https://doi.org/10.1152/physrev.1992.72.1.165
  29. Methods Mol Biol v.120 Determination of phospholipase A2s expression in mast cells by reverse-transcriptase polymerase chain reaction Fonteh, A.N.;Reed, W.;Samet, J.M.
  30. Prostaglandins v.16 no.2 Effect of honey bee venom on prostaglandin levels in mouse skin Schmidt, D.K.;Destephano, D.B.;Brady, U.E. https://doi.org/10.1016/0090-6980(78)90024-2
  31. Brain Res. v.171 Microinjection of morphine into the periaqueductal gray evokes the release of serotonin from spinal cord Yaksh, T.L.;Tyce, G.M. https://doi.org/10.1016/0006-8993(79)90747-9
  32. Allergy Clin. Immunol. v.101 no.6 Successful immunotherapy with T-cell epitope peptides of bee venom phospholipase A2 induces specific T-cell anergy in patients allergic to bee venom Muller, U.;Akdis, C.A.;Fricker, M.;Akdis, M.;Blesken, T.;Bettens, F.;Blaser, K. https://doi.org/10.1016/S0091-6749(98)70402-6
  33. Int. Arch. Allergy Immunol v.117 no.1 Determinants and mechanisms of human immune responses to bee venom phospholipase A2 Blaser, K.;Carballido, J.;Faith, A.;Crameri, R.;Akdis, C.
  34. 중국중서의결합잡지 v.13 no.4 Comparison of anti-inflammatory, analgesic activities, anaphylactogenicity and acute toxicity between bee venom and its peptides Chen, C.Y.;Chen, W.X.;Sun, X.
  35. Agents Actions v.9 no.2 Anti-arthritic effect of bee venom Chang, Y.H.;Bliven, M.L. https://doi.org/10.1007/BF02024736
  36. Drug Topics Serotonin: The Neurotransmitter for the '90s Ronald F.Borne
  37. Science v.230 no.4732 Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia Saike, R.K.;Scharf, S.;Faloona, F.;Mullis, K.B.;Horn, G.T.;Erlich, H.A.;Arnheim, N. https://doi.org/10.1126/science.2999980
  38. Science v.233 no.4768 Direct cloning and sequence analysis of enzymatically amplified genomic sequences Scharf, S.J.;Horn, G.T.;Erlich, H.A. https://doi.org/10.1126/science.3461561
  39. Science v.239 no.4839 Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase Saike, R.K.;Gelfand, D.H.;Stoffel, S.;Scharf, S.J.;Higuchi, R.;Horn, G.T.;Mullis, K.B.;Erlich, H.A. https://doi.org/10.1126/science.2448875
  40. Journal of Biological Chemistry v.264 no.11 Isolation, characterization, and expression in Escherichia coli of the DNA polymerase gene from Thermus aquaticus Lawyer, F.C.;Stoffel, S.;Saiki, R.K.;Myambo, K.;Drummond, R.;Gelfand, D.H.
  41. Life Sci. v.25 Electro-acupuncture analgesia could be mediated by at least two pain relieving mechanisms: Endorphin and non endorphin systems Cheng Rss;Pomeranz, B. https://doi.org/10.1016/0024-3205(79)90598-8
  42. Sci. Sin. v.22 the role of central 5-hydroxytryptamine in acupuncture analgesia Han, J.S.;Chou, P.H.;Lu, Z.C.;Yang, T.H.(et al.)
  43. International Journal of neuroscience v.29 A mesolimbic loop of analgesia. I. Activation by morphine of a serotonergic pathway from periaqueductal gray to nucleus accumbens Han, J.S.;Xuan, Y.T. https://doi.org/10.3109/00207458608985641
  44. Molecular Neurobiology of the mammlian Brain McGeer, P.L.;Eccles, J.C.;McGeer, E.G.
  45. Am. J. Chinese Med. v.2 Human medical college: The relation between Acupuncture, Analgesia and neurotransmitters in Rabbit brain Acupuncture Analgesia Research group
  46. JAP. J. Pharmacol. v.49 no.2 Further evidence for possible analgesic mechanism of electroacupuncture: Effects on neuropeptides and serotonergic neurons in rat spinal cord Tsai, H.Y.;Lin, J.G.;Inoki, R. https://doi.org/10.1254/jjp.49.181
  47. Brain Res. Rev. v.10 Is the Jaw-Opening Reflex a Valid Model of Pain? Mason, P.;Strassman, A.;Maciewicz, R. https://doi.org/10.1016/0165-0173(85)90003-7
  48. Experimental neurology v.93 Is Electrical stimulation of the Rat Incisor an Appropriate Experimental Nociceptive Stimulus? Rajaona, J.;Dallel, R.;Woda, A. https://doi.org/10.1016/0014-4886(86)90190-1
  49. Pain v.15 Is a selective stimulation of the rat incisor tooth pulppossible? Engstrand, P.;Shyu, B.C.;Andersson, S.A. https://doi.org/10.1016/0304-3959(83)90004-0
  50. Exp Neurol v.67 A problem in electrical stimulation of incisor tooth pulp in rats Hayashi, H. https://doi.org/10.1016/0014-4886(80)90243-5
  51. Jap. J. Physiol. v.28 Effects of electroacupuncture on rat jaw opening reflex elicited by tooth pulp stimulation Kazuo Toda https://doi.org/10.2170/jjphysiol.28.485
  52. Arch. unt. Pharmacodyn. v.270 Variations of the Jaw Opening Reflex Observed in Awake, Freely Moving Rats Pajot, J.;Vassel, A.;Aigouy, L.;Rajaona, J.;Woda, A.
  53. Prostanglandins Other Lipid Mediat v.57 no.5-6 Contribution of type II PLA2 to prostaglandin formation: a study using a type II PLA2 specific inhibitor SB 203347 Munns, M.J.;King, R.G.;Rice, G.E. https://doi.org/10.1016/S0090-6980(99)00012-X