[논문]Collagen-induced Arthritis Rat Model에서 염증성 통증에 대한 봉독약침의 진통효과 및 기전연구: 5HT-3 & Muscarinic Cholinergic Mechanisms에 대한 연구

서병관; 박동석; 백용현

[국내논문] Collagen-induced Arthritis Rat Model에서 염증성 통증에 대한 봉독약침의 진통효과 및 기전연구: 5HT-3 & Muscarinic Cholinergic Mechanisms에 대한 연구
Antinociceptive Effect and the Mechanism of Bee Venom Pharmacopuncture on Inflammatory Pain in the Rat Model of Collagen-induced Arthritis: Mediation by 5HT-3 & Muscarinic Cholinergic Receptors 원문보기

大韓鍼灸學會誌= The journal of Korean Acupuncture & Moxibustion Society, v.28 no.1, 2011년, pp.37 - 44

서병관 (경희대학교 한의과대학 침구학교실) , 박동석 (경희대학교 한의과대학 침구학교실) , 백용현 (경희대학교 한의과대학 침구학교실)

초록
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배경 및 목적 : 봉독약침요법(bee venom pharmacopuncture, BVP)은 rheumatoid arthritis(RA)의 치료에 활용되고 있으나, RA로 인한 염증성 통증에 대한 봉독약침의 진통효과와 specific mechanism은 아직까지 명확하게 밝혀지지 않았다. 이에 본 연구에서는 RA animal model로서 collagen-induced arthritis(CIA) rat model에서 봉독약침의 a1-adrenergic, 5HT-3 그리고 muscarinic cholinergic mechanism을 확인하고자 한다. 방법 : CIA를 유도하기 위하여 male Sprague-Dawley rat에 freund's incomplete adjuvant에 유화(乳化)시킨 bovine type II collagen을 주입하고 14일 후 booster injection 시행하였다. 진통효과는 tail flick latency (TFL)로 평가하였다. 결과 : 관절염의 유도 이후 염증성 통증 역치는 시간이 지나면서 낮아지며, 5주 이후로는 통증 역치에 큰 변화가 없이 유지되었다. 첫 번째 immunization으로부터 5주 경과 후 족삼리($ST_{36}$)에 봉독약침처치(0.25 mg/ kg)를 시행하여 유의한 진통효과를 관찰하였다. 또한 봉독약침의 진통효과는 ondansetron(5HT-3 receptor antagonist, 0.5mg/kg, i.p.), atropine(muscarinic cholinergic receptor antagonist, 1mg/kg, i.p.)의 전처치에 의하여 억제되었으나, prazosin(a1-adrenergic receptor antagonist, 1mg/kg, i.p.)의 전처치에 의해서는 억제되지 않았다. 결론 : 봉독약침은 CIA로 인한 염증성 통증에 유의한 진통효과를 나타내며 그 analgesic mechanism은 5HT-3와 muscarinic cholinergic receptor에 의하여 매개되며 a1-adrenergic receptor에 의하여 매개되지는 않았다.

Keyword

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문제 정의

While Bee Venom Pharmacopuncture (BVP) shows the analgesic effect in RA, the antinociceptive mechanisms related with the inflammatory pain by using the bovine type II collagen-induced RA model have not been fully studied. The primary purpose of this study was to determine whether Bee Venom Pharmacopuncture (BVP) is able to show the antinociceptive effect on inflammatory pain in the rat model of collagen-induced arthritis (CIA). A second goal of this study was to clarify whether the antinociceptive effect of BVP is related to the activation of α1-adrenoceptor, 5HT-3 and muscarinic cholinergic receptors.

가설 설정

Several types of acupoint stimulation techniques including electroacupuncture, Bee venom pharmacopuncture, moxibustion and acupressure have been used in order to produce antinociceptive effects that are selectively mediated by the endogenous descending modulatory systems ^20-22). In this study, we intended to examine the relation with monoaminergic systems including adrenalin, serotonin and acetylcholine. So, to reveal the antinociceptive effect of BVP on the inflammatory pain induced by CIA, agonists and antagonists of α1-adrenoceptor, 5HT-3 and muscarinic cholinergic receptors were pretreated before the initiation of BVP.

제안 방법

A second goal of this study was to clarify whether the antinociceptive effect of BVP is related to the activation of α1-adrenoceptor, 5HT-3 and muscarinic cholinergic receptors.
In order to reveal the related analgesic mechanism, the selective α1-adrenoceptor agonist phenylephrine (ICN Biomedicals, Ohio, USA; 2mg/kg, i.p.) and antagonist prazosin (ICN Biomedicals, Ohio, USA; 1mg/kg, i.p.), 5HT-3 agonist m-chlorophenyl-biguanide (ICN Biomedicals, Ohio, USA; 1mg/kg, i.p.) and antagonist ondansetron (ICN Biomedicals, Ohio, USA; 0.5mg/kg, i.p.), muscarinic cholinergic agonist neostigmine (ICN Biomedicals, Ohio, USA; 100㎍g/kg, i.p.) and antagonist atropine (ICN Biomedicals, Ohio, USA; 1mg/kg, i.p.) were injected intraperitoneally 20 minutes before BVP treatment.
5 weeks after the first administration of bovine type II collagen, the behavioral test was performed with the tail flick unit prior to (as baseline test) and 10, 20, 30, 45 and 60min after BVP treatment.

데이터처리

The significance of statistical differences were determined using non-parametric Friedman’s rank test followed by Dunnett’s posthoc test in a group, non-parametric Mann-Whitney U-test between two groups and non-parametric Kruskal-Wallis ANOVA followed by Dunnett’s posthoc test among groups.

이론/모형

The room was light/dark (08 : 00～20 : 00h light, 20 : 00～08 : 00h dark) controlled and kept at 2124℃. All experiments were conducted in accordance with the guidelines of the International Association for the Study of Pain (IASP)¹²⁾.

성능/효과

5. There were statistically significant differences in TFL between the antagonist atropine pretreatment Zusanli BVP treatment group (BVP+Atrop, n=10) and the DMSO pretreatment Zusanli BVP treatment group (BVP+DMSO, n=10) at 10, 20, 30 and 45min after the initiation of BVP.
4. There were statistically significant differences in TFL between the antagonist ondansetron pretreatment Zusanli BVP treatment group (BVP+ondan, n=10) and the DMSO pretreatment Zusanli BVP treatment group (BVP+DMSO, n=10) at 10, 20, 30 and 45min after the initiation of BVP. This results show that the 5HT-3 receptor antagonist ondansetron pretreatment significantly blocked the BVP antinociceptive effects.

후속연구

To examine the whole mechanism related with BVP antinociception in chronic inflammatory pain induced by RA, further study should be done in relation with other types of opioid receptors (δ-, κ-), serotonergic receptor (5HT-4) and nicotinic cholinergic receptor.
To examine the whole mechanism related with BVP antinociception in chronic inflammatory pain induced by RA, further study should be done in relation with other types of opioid receptors (δ-, κ-), serotonergic receptor (5HT-4) and nicotinic cholinergic receptor. And, in relation with the interactive effects among receptors on BVP-induced antinociception, we also think the further study is needed to confirm this fact.

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