PUBLICATION
Identification of cryptotanshinone from Tongmai to inhibit thrombosis in zebrafish via regulating oxidative stress and coagulation cascade
- Authors
- Sheng, J., Meng, Q., Yang, Z., Guan, J., Zhao, Y., Zhang, J., Wang, Y., Zhao, L., Wang, Y.
- ID
- ZDB-PUB-200621-7
- Date
- 2020
- Source
- Phytomedicine : international journal of phytotherapy and phytopharmacology 76: 153263 (Journal)
- Registered Authors
- Keywords
- Coagulation cascade, Cryptotanshinone, Oxidative stress, Phenylhydrazine, Thrombosis, Zebrafish
- MeSH Terms
- none
- PubMed
- 32563016 Full text @ Phytomedicine
Citation
Sheng, J., Meng, Q., Yang, Z., Guan, J., Zhao, Y., Zhang, J., Wang, Y., Zhao, L., Wang, Y. (2020) Identification of cryptotanshinone from Tongmai to inhibit thrombosis in zebrafish via regulating oxidative stress and coagulation cascade. Phytomedicine : international journal of phytotherapy and phytopharmacology. 76:153263.
Abstract
Background Thromboembolic events are leading causes of mortality and morbidity all over the world. Tongmai (TM) is a botanical drug with valid clinical efficacy and safety in the management of thrombosis and ischemic cardiovascular diseases, however, its active compounds and underlying mechanism are largely unclear.
Purpose To investigate the endogenous effects, therapeutic mechanism and active compounds of TM in thrombus formation.
Study design Combined with transgenic zebrafish models and high-content imaging system, this study evaluated the endogenous antithrombotic effects of TM and screened for the active compounds.
Methods The PHZ-induced thrombotic model in erythrocytes or platelets labeled transgenic zebrafish were established, to dynamically evaluate the antithrombotic effects of TM. The oxidative damage levels were analyzed by specific fluorescent probes, and the expression levels of key factors in coagulation cascades and platelet activation were examined by QPCR. TM were dissected into fractions by reverse phase chromatography and subsequently screened for their antithrombotic effects in the transgenic fish models. The compounds of the active TM fraction were then analyzed by UPLC-Q-TOF analysis and further verified for their antithrombotic effects and mechanisms.
Results In PHZ-induced zebrafish thrombotic model, TM incubation markedly increased cardiac blood flow, decreased peripheral erythrocytes aggregation, and recovered peripheral platelet circulation. Besides, the levels of oxidative stress and lipid peroxidation were increased in the PHZ-induced thrombotic fish, which were greatly decreased by TM treatment. Moreover, TM significantly reduced the expression of coagulation factor II (thrombin) and the downstream fibrinogen. In order to identify the active compounds of TM, four fractions were separated from the extract by reverse phase chromatography, which were subsequently screened for their antithrombotic effects in the fish model. As a result, fraction 4 showed the strongest effect in inhibiting thrombosis. Finally, through UPLC-Q-TOF analysis and endogenous screening, cryptotanshione was identified as the main active compound with antithrombotic effects.
Conclusion Our study demonstrated the endogenous antithrombotic effects of TM, which is possibly mediated by inhibiting oxidative stress and coagulation cascade. Cryptotanshione was identified as a major compound with antithrombotic activity and is a promising candidate for novel antithrombotic therapy.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping