皖南地区眼镜蛇毒镇痛组分对大鼠炎性痛作用机制探讨

中国临床药理学与治疗学 ›› 2013, Vol. 18 ›› Issue (10): 1093-1099.

皖南地区眼镜蛇毒镇痛组分对大鼠炎性痛作用机制探讨

王海华^1,2, 张根葆², 闵志雪^1,2, 黄璐², 崔凤娟^1,2

¹皖南医学院生理教研室,
²蛇毒研究所,芜湖 241002,安徽

收稿日期:2013-05-06 修回日期:2013-09-10 出版日期:2013-10-26 发布日期:2013-09-30
通讯作者: 张根葆,男,教授,硕士生导师,研究方向:心血管病理生理学,蛇毒毒理学。Tel: 0553-0932435 E-mail: zgb858@163.com
作者简介:王海华,男,副教授,硕士生导师,研究方向:痛觉及其调制、心血管病理生理学。Tel: 13675536187 E-mail: wanghaihua9972@sina.com
基金资助:
安徽省自然科学研究重点项目(KJ2013A251);皖南医学院重点科研项目培育基金(WK2012Z01);活性生物大分子研究安徽省重点实验室(科基[2012]126号)

Study on analgesic effect of active component from crude cobra venom in Wannan Area

WANG Hai-hua^1,2, ZHANG Gen-bao², MIN Zhi-xue^1,2, HUANG Lu², CUI Feng-juan^1,2

¹Department of Physiology,
²Institute of snake venom,Wannan Medical College,Wuhu 241002, Anhui, China

Received:2013-05-06 Revised:2013-09-10 Online:2013-10-26 Published:2013-09-30

摘要/Abstract

摘要： 目的:探讨皖南地区眼镜蛇毒镇痛组分(CVAF)对大鼠炎性痛作用效应及其可能机制。方法:40只雄性SD大鼠在测定基础痛阈后随机分为4组(n=10):正常对照组(NC 组)、炎性痛模型组(CFA 组)、炎性痛模型+CVAF组(CFA+CVAF 组)和炎性痛模型+吗啡组(CFA +Morphine 组)。随后将36只SD雄性大鼠随机分成6组(n=6):正常对照组(NC 组)、炎性痛模型组(CFA 组)、炎性痛模型+CVAF组(CFA+CVAF 组)、炎性痛模型+阿托品＋CVAF组(CFA+Atr＋CVAF)、炎性痛模型+纳洛酮＋CVAF组(CFA+Nal＋CVAF)和炎性痛模型+L-NAME＋CVAF组(CFA+L-NAME＋CVAF)。测定各组大鼠的热痛阈潜伏期(TWL)和机械痛阈值(MWT),制备脊髓匀浆,用于检测其中IL-1和TNF-α的表达。结果:与NC组相比,CFA组大鼠第1天开始TWL和MWT明显降低,出现热痛觉过敏和机械痛觉过敏,持续 14 d 仍存在;CFA组大鼠脊髓匀浆IL-1和TNF-α表达均明显增高(P<0.01)。与CFA组相比,CFA组大鼠腹腔注射CVAF后其TWL和MWT均升高,表明炎性痛大鼠热痛觉过敏和机械痛觉过敏得到改善;脊髓匀浆中IL-1、TNF-α表达显著减少(P<0.01),显示CVAF通过抑制炎性因子的产生而发挥镇痛作用。与CFA+CVAF组大鼠相比,L-NAME+CVAF组大鼠明显增强了CVAF对炎性痛大鼠的镇痛作用,表现为TWL和MWT的明显升高(P<0.01);而CFA+Atr＋CVAF组和CFA+Nal＋CVAF组大鼠TWL和MWT均有不同程度的降低(P<0.01)。结论:初步阐明CVAF通过减少炎性因子的释放对炎性痛大鼠具有镇痛效应。阿片肽受体系统和胆碱能受体系统均部分参与了其对炎性痛大鼠的镇痛作用。

关键词: 眼镜蛇毒, 分离纯化, 大鼠, 镇痛机制

Abstract: AIM: To study on analgesic effect of active component from crude cobra venom in Wannan Area(CVAF) and investigate its possible mechanism of analgesic effect.METHODS: Inflammatory pain model of rat (CFA group) was established using CFA (150 μL, s.c.). 40 male SD rats were randomly divided into 4 groups (n=10): NC, CFA, CFA+CVAF and CFA+Morphine groups. Then another 36 male SD rats were randomly divided into 6 groups (n=6): NC, CFA, CFA+CVAF, CFA+Atr+CVAF, CFA+Nal+CVAF and CFA+L-NAME+CVAF groups, the changes of thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT), induced by CVAF, were determined by Spurs pain and mechanical pressure methods, respectively. The levels of interleukin (IL)-1 and tumor necrosis factor (TNF)-α in spinal cord were detected by enzyme-linked immunosorbent assay (ELISA). Analgesic activity of CVAF was also evaluated in the CFA group after administrating with atropine, naloxone or L-NAME.RESULTS: Compared with NC group, TWL and MWT were decreased from 1st day to 14th day in CFA group; the levels of IL-1 and TNF-α from spinal cord in CFA group were significantly increased (P<0.01). Compared with the CFA group,the TWL and MWT were increased after injecting CVAF,thermal hyperalgesia and mechanical allodynia were also improved simultaneously,the levels of IL-1 and TNF-α were decreased (P<0.01), the result showed CVAF played a role in analgesic activity via inhibition of inflammatory factor.Compared with CFA+CVAF group, atropine or naloxone could return the increased TWL and MWT by CVAF (P<0.01). The results indicated that both opioid peptide receptor system and cholinergic receptor system could be related to analgesic effect of CVAF for inflammatory pain.CONCLUSION: CVAF plays a role in analgesic effect with inflammatory pain through inhibition of inflammatory factor. Both opioid peptide receptor system and cholinergic receptor system are partially involved in the analgesic effect of CVAF for inflammatory pain.

Key words: Cobra venom, Separation purification, Rat, Analgesic mechanism

中图分类号:

R965.2

王海华, 张根葆, 闵志雪, 黄璐, 崔凤娟. 皖南地区眼镜蛇毒镇痛组分对大鼠炎性痛作用机制探讨[J]. 中国临床药理学与治疗学, 2013, 18(10): 1093-1099.

WANG Hai-hua, ZHANG Gen-bao, MIN Zhi-xue, HUANG Lu, CUI Feng-juan. Study on analgesic effect of active component from crude cobra venom in Wannan Area[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2013, 18(10): 1093-1099.

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