Rapid Enrichment and Activity Research of Antithrombotic Proteins from Pheretima vulgaris
10.13422/j.cnki.syfjx.20211562
- VernacularTitle:通俗环毛蚓抗血栓活性蛋白成分的快速富集及活性考察
- Author:
Jing-wei ZHANG
1
;
Wan-qing YANG
1
;
Yun-nan MA
1
;
Wen-jie WANG
1
;
Wan-ling ZHONG
1
;
Jia-yao BI
1
;
Shou-ying DU
1
Author Information
1. School of Chinese Materia Medica，Beijing University of Chinese Medicine，Beijing 100029，China
- Publication Type:Research Article
- Keywords:
Pheretima;
Pheretima vulgaris;
anticoagulant activity;
fibrinolytic activity;
protein purification;
temperature;
antithrombotic activity
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(22):166-171
- CountryChina
- Language:Chinese
-
Abstract:
Objective:A strong antithrombotic protein component， named PvQ， was purified and enriched from total protein of Pheretima vulgaris， a traditional Chinese medicine. Moreover， we evaluated its fibrinolytic and anticoagulant activity， and expected to provide reference for the research on antithrombotic substances of Pheretima. Method:A rapid in vitro activity-oriented separation combined with the AKTA-Pure protein purification system conducted on P. vulgaris. Meanwhile， the fibrinolytic and anticoagulant activities of PvQ were measured by fibrin plate method and fibrinogen-thrombin time （Fibg-TT） method. And the in vitro thrombolysis assay was used for evaluating the lysis ability of PvQ to thrombus. Then the stability of PvQ was also analyzed for its anticoagulant activity at different pH and temperature. Result:The PvQ was successfully enriched and its activity was determined to have significant fibrinolytic and anticoagulant activities. And the result of in vitro thrombolysis assay revealed that PvQ could hydrolyze more than 80% of thrombus after 5 h of incubation at 37 ℃. In addition， the changes of temperature and pH had significant effects on antithrombotic activity， and this study showed that PvQ was rapidly inactivated at ≥60 ℃ or in acidic conditions （pH<7）. While， the activity of PvQ was unaffected or less affected at ≤50 ℃ and under alkaline conditions. Conclusion:A feasible preparation method of PvQ is established， and it can affect fibrin and fibrinogen at the same time， thus exerting a dual fibrinolytic effect and possessing significant fibrinolytic and anticoagulant activities. It provides a scientific interpretation for the treatment of thrombotic diseases by PvQ and a reference for the development of antithrombotic protein products of Pheretima.
