森林山蛭（Haemadipsa sylvestris）中两个天然组分的纯化、结构及生物活性研究
文献类型：学位论文
| 作者 | 龙承波 |
| 学位类别 | 硕士 |
| 答辩日期 | 2015-05 |
| 授予单位 | 中国科学院研究生院 |
| 授予地点 | 北京 |
| 导师 | 赖仞 |
| 关键词 | 森林山蛭 TTX-R钠离子通道 PGE1 血小板 |
| 其他题名 | Purification, structure and biological activities of two natural components from leech Haemadipsa sylvestris |
| 学位专业 | 生物工程 |
| 中文摘要 | 吸血蛭类在医疗领域的应用历史悠久。西方水蛭疗法最早记载见于埃及法老墓壁画（公元前1500）和印度梵文（公元前1300）中。我国药用蛭记载最早见于《神农本草经》。吸血蛭类具有抗凝血、镇痛和炎症调节等作用。现今水蛭的两种主要用法为直接运用活体水蛭的水蛭疗法和以干水蛭为配伍药的中成药。水蛭疗法主要流行于西方国家，而干水蛭入药的用法主要是在中国。 临床治疗效果研究和观察发现吸血蛭类在抗血栓、镇痛和炎症调节等方面的治疗效果比常规药物治疗效果好很多。但目前还没有关于镇痛或炎症调节活性的单一成分的报道，抗凝血成分也仅仅局限于多肽蛋白类。由于缺乏对这些生物学活性的物质基础的研究，在很大程度上限制了吸血蛭类的开发和应用。森林山蛭（Haemadipsa sylvestris）具有个体小，吸血速度快，抗凝血、镇痛和炎症调节作用强的特点。本论文以此为切入点，通过各种分离纯化手段对森林山蛭具有抗凝血、镇痛和炎症调节作用的组分进行跟踪纯化。 首次从森林山蛭中分离纯化到具有选择性抑制TTX-R钠离子通道抑制活性的多肽HSTX-Ⅰ。其对大鼠DRG上TTX-R钠离子通道抑制作用IC50为15.13±2.40 μM。通过同源扩增的方法还从山蛭头部cDNA文库中克隆到13条与HSTX-Ⅰ同源的序列，命名为HSTX-II—HSTX-XIV。在NCBI数据库中比对，未发现任何同源序列。因此推测该类多肽为新的多肽家族。由于TTX-R钠离子通道和疼痛相关，推测在山蛭取食宿主血液时，HSTX-Ⅰ可能会在一定程度上抑制宿主的疼痛反应。 同样，我们也是首次从山蛭体内分离纯化到具有多种生物学功能的PGE1。其对5 μM ADP诱导的血小板聚集抑制作用的IC50为21.81±2.24 nM。PGE1普遍存在于哺乳动物体内，其具有扩张血管、调节炎症反应和抑制血小板聚集的作用，具此推测山蛭很可能通过利用PGE1来帮助其从宿主吸血。 |
| 英文摘要 | Leech therapy has a long and widespread tradition, as Pharaonic tomb paintings (1500 BC) and Sanskrit writings (1300 BC). Record of medicinal leech was first seen in "Shen Nong Ben Cao Jing" in China. Anticoagulation, analgesic and inflammation regulation are mainly functions of bloodsucking leeches. Today, live leeches used in hirudotherapy and medicinal materials are largely bloodsucking leeches. Hirudotherapy is popular mainly in western countries, and as medicinal material usage mainly in China. Clinical observations showed that bloodsucking leech therapy were better than conventional therapies on anticoagulation, analgesic and inflammation regulation. But with no report of a single component having analgesic and regulation of inflammation and only a few anticoagulant polypeptide /proteins discovered, the application of blood-sucking leeches is greatly limited. H. sylvestris is small and sucks blood rapidly with efficacy features of anticoagulant, analgesic and regulation of inflammation. In this paper, regarding these points, we tracked the activities along with a variety of purification methods. We first isolated a novel peptide from H.sylvestris. It could selectively inhibit the tetrodotoxin-resistive(TTX-R) sodium channels in rat dorsal root ganglion(DRG) with an IC50 of 15.13±2.40 μM. Thirteen homologous sequences of HSTX-Ⅰ, named as HSTX-II to HSTX-XIV, were cloned from the cDNA library of H.sylvestris. Blast search did not find any homologous sequence, suggesting that these sequences belong to a new polypeptide family. Because TTX-R sodium channels are often associated with pain, HSTX-Ⅰmay inhibit pain response of the host when H. sylvestris feeds on blood. In addition, we isolated PGE1 from H. sylvestris. PGE1 has many biological functions. The purified PGE1 showed strong ability to inhibit platelet aggregation induced by ADP and the IC50 was 21.81 ± 2.24 nM. PGE1 was commonly found in mammals, which can dilate vessels, regulate inflammation and inhibit platelet aggregation. PGE1 may facilitate H. sylvestris to get blood meals from their hosts. |
| 语种 | 中文 |
| 源URL | [http://159.226.149.26:8080/handle/152453/10198]  |
| 专题 | 昆明动物研究所_动物毒素室 |
| 推荐引用方式 GB/T 7714 | 龙承波. 森林山蛭（Haemadipsa sylvestris）中两个天然组分的纯化、结构及生物活性研究[D]. 北京. 中国科学院研究生院. 2015. |
入库方式： OAI收割
来源：昆明动物研究所
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