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银环蛇蛇毒中具有镇痛活性的多肽成分的生化分离和活性鉴定*
Biochemical isolation and activity identification of polypeptide components with analgesic activity in Bungarus multicinctus Venom
投稿时间:2019-02-27  修订日期:2019-04-08
DOI:
中文关键词:  银环蛇  银环蛇毒  纯化  镇痛
英文关键词:Bungarus multicinctus  Bungarus multicinctus venom  Purification  Analgesic effect
基金项目:
作者单位E-mail
刘金满 广州中医药大学临床药理研究所 广州 liujinman77@163.com 
卢悟广 南京中医药大学附属中西医结合医院  
陈永根 广州中医药大学临床药理研究所 广州  
王明远 南京中医药大学附属中西医结合医院  
王奇 广州中医药大学临床药理研究所 wqitcm@qq.com 
曹鹏 南京中医药大学附属中西医结合医院  
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中文摘要:
      目的:研究银环蛇(Bungarus multicinctus)毒的镇痛活性,并对其中的活性单体进行分离、纯化与鉴定。方法:采用热痛、醋酸扭体动物模型检测银环蛇(Bungarus multicinctus)蛇毒毒的镇痛作用,通过分子筛、高效液相(HPLC)等纯化手段从银环蛇毒中分离出镇痛单体,然后通过肽谱鉴定,氨基酸测序及序列比对鉴定其结构特征。结果:本研究发现银环蛇粗毒具有显著的镇热痛活性,与生理盐水组相比,热痛阈提高近2℃。与吗啡相比,其镇痛作用持续时间更长,在给药7h后仍有显著镇痛效果。通过分子筛及高效液相分离,我们发现其镇痛活性成分主要为小分子多肽,其中活性最强的成分是α-bungarotoxin。α-bungarotoxin有与银环蛇粗毒类似的热镇痛活性,并且能够显著抑制醋酸导致的急性炎性痛。序列鉴定及结构分析表明α-bungarotoxin是一种含有74个氨基酸,5对二硫键的三指肽,能够与乙酰胆碱受体结合。结论:银环蛇(Bungarus multicinctus)蛇毒具有显著的镇痛作用。其中作用于乙酰胆碱受体的三指肽α-bungarotoxin是其最主要的镇痛活性成分。
英文摘要:
      OBJECTIVE: This research was performed to study the analgesic activity of Bungarus multicinctus and isolation, purification as well as identification of the active component. METHODS: The analgesic effect of Bungarus multicinctus on mouse was detected by using published heat pain and acetic acid inflammatory pain mouse models. The Bungarus multicinctus venom was purified by a series section, for example, Molecular exclusion and High Performance Liquid Chromatography (HPLC). Analgesic components were isolated from Bungarus multicinctus venom and its structural features identified by peptide mapping, comparison amino acid sequences with standard. RESULTS: we found that compared with the saline group, the Bungarus multicinctus venom increased 2℃ at the thermal pain threshold test with a very low concentration, and it remains effective analgesic effect after 7 hours of administration, which is much longer than morphine. We further seperate one of the Bungarus multicinctus venom components G50 - P4 by a series section, for example, Molecular exclusion and HPLC and We found that the analgesic active components is mainly comprised of small molecule polypeptides , and the most active one is α-bungarotoxin, which has a similar thermal analgesic activity as the Bungarus multicinctus, and acute inflammatory pain induced by acetic acid can be significantly inhibited. By peptide mapping and amino acid sequence comparison,we found that α- bungarotoxin is a typical three-finger peptide, containing 74 amino acids and 5 pairs of disulfide bonds, and it is capable of binding to acetylcholine receptors were been clarified. CONCLUSION: Bungarus multicinctus venom has impressed analgesic effect, the three-finger peptide α-bungarotoxin, acting on the acetylcholine receptor is the main analgesic active component.
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