Zheng Xiaobei, Li Juan, Chen Zhengwang, Liu Yimei, Chen Keli
Department of Identification and Assessment of TCM, Hubei University of Traditional Chinese Medicine, Key Laboratory of TCM Resource and TCM Compound Co-constructed by Hubei province and Ministry of Education, New products of TCM Senile Diseases Co-Innovation Center of Hubei ; Department of Quality Control, Wuhan Institute of Biological Products Co., Ltd., China.
Department of Identification and Assessment of TCM, Hubei University of Traditional Chinese Medicine, Key Laboratory of TCM Resource and TCM Compound Co-constructed by Hubei province and Ministry of Education, New products of TCM Senile Diseases Co-Innovation Center of Hubei.
Pharmacogn Mag. 2015 Jul-Sep;11(43):444-8. doi: 10.4103/0973-1296.160446.
Dried Whitmania pigra is used for the treatment of cardiovascular and cerebrovascular diseases in traditional Chinese medicine and hot water and alcohol extracts also have anticogulant activity. However, a lower molecular weight and more stable anticogulant is needed.
The objective of the following study is to purify and characterize of an anticoagulant oligopeptide from Hirudo (Whitmania pigra Whitman).
Gel filtration on Sephadex G-50, ion exchange on diethylaminoethyl-cellulose, and semi-prepared high-performance liquid chromatography were used to purify Hirudo. Automated coagulation analyzer was used for evaluating anticoagulant activity. Molecular weight was measured by Matrix-assisted laser desorption ionization time of flight mass spectrometry. Amino acid sequence of the oligopeptide was measured by amino acid sequence analyzer.
A new anticoagulant, named whitide, isolated from Hirudo was purified, with a molecular weight 1997.1 Da. Amino acid sequence of the oligopeptide was identified as Gly-Pro-ALa-Gly-Hyp-Val-Gly-Ala-Hyp-Gly-Gly-Hyp-Gly-Val-Arg-Gly-Leu-Hyp-Gly-Asp-Arg-Gly. The results revealed that its amino acid sequence had strong homology to various types of collagen.
Whitide might be an orally anticoagulant for its hot and trypsin stable.
水蛭干在传统中药中用于治疗心脑血管疾病，其热水提取物和乙醇提取物也具有抗凝活性。然而，需要一种分子量更低且更稳定的抗凝剂。
以下研究的目的是从水蛭（宽体金线蛭）中纯化并鉴定一种抗凝寡肽。
采用Sephadex G - 50凝胶过滤、二乙氨基乙基纤维素离子交换和半制备高效液相色谱法纯化水蛭。使用自动凝血分析仪评估抗凝活性。通过基质辅助激光解吸电离飞行时间质谱法测定分子量。用氨基酸序列分析仪测定寡肽的氨基酸序列。
从水蛭中分离出一种新的抗凝剂，命名为水蛭肽，其分子量为1997.1 Da。该寡肽的氨基酸序列被鉴定为甘氨酸 - 脯氨酸 - 丙氨酸 - 甘氨酸 - 羟脯氨酸 - 缬氨酸 - 甘氨酸 - 丙氨酸 - 羟脯氨酸 - 甘氨酸 - 甘氨酸 - 羟脯氨酸 - 甘氨酸 - 缬氨酸 - 精氨酸 - 甘氨酸 - 亮氨酸 - 羟脯氨酸 - 甘氨酸 - 天冬氨酸 - 精氨酸 - 甘氨酸。结果显示其氨基酸序列与各种类型的胶原蛋白具有高度同源性。
水蛭肽因其热稳定性和对胰蛋白酶的稳定性，可能是一种口服抗凝剂。
