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Updated: Dec 9, 2025
Ferric Chloride-induced Murine Thrombosis Models
Published on: September 5, 2016
AMPK inhibition protects against arterial thrombosis while sparing hemostasis through differential modulation of
Paresh P Kulkarni1, Vijay K Sonkar2, Deepa Gautam1
1Centre for Advanced Research on Platelet Signaling and Thrombosis Biology (ICMR), Department of Biochemistry, Institute of Medical Sciences, Banaras Hindu University, Varanasi, Uttar Pradesh, India.
Abstract:
AMP-activated protein kinase (AMPK) is a metabolic master switch that has critical role in wide range of pathologies including cardiovascular disorders. As AMPK-α2 knockout mice exhibit impaired thrombus stability, we asked whether pharmacological inhibition of AMPK with a specific small-molecule inhibitor, compound C, could protect against arterial thrombosis without affecting hemostasis. Mice pre-administered with compound C exhibited decreased mesenteric arteriolar thrombosis but normal tail bleeding time compared to vehicle-treated animals. Compound C potently restricted platelet aggregation, clot retraction and integrin activation induced by thrombin and collagen. It impaired platelet spreading on both immobilized fibrinogen and collagen matrices; it, however, had no significant effect on thrombin-induced phosphatidylserine exposure that is characteristic of procoagulant platelets. In parallel, compound C brought about significant drop in thrombin-induced phosphorylation of myosin light chain (MLC) and MLC phosphatase (MYPT1) as well as abrogated rise in level of RhoA-GTP in thrombin-stimulated platelets. Thus, effects of compound C on agonist-induced platelet responses could be at least in part attributed to modulation of cytoskeletal changes mediated by RhoA-MYPT1-MLC signaling. An ideal antithrombotic drug would spare hemostatic responses that maintain vascular integrity while preferentially protecting against thrombosis. The present study suggests that AMPK could be one such potential therapeutic target.
Insights
Pharmacological inhibition of AMP-activated protein kinase (AMPK) with compound C reduced arterial thrombosis in mice without impacting hemostasis. This suggests AMPK is a potential therapeutic target for antithrombotic drugs.
Area of Science:
- Biochemistry
- Pharmacology
- Cardiovascular Research
Background:
- AMP-activated protein kinase (AMPK) is a key metabolic regulator implicated in various pathologies, including cardiovascular disorders.
- AMPK-α2 knockout mice show compromised thrombus stability, indicating a role in thrombosis.
Purpose of the Study:
- To investigate if pharmacological inhibition of AMPK using compound C could offer protection against arterial thrombosis.
- To determine if AMPK inhibition affects hemostasis, the process that stops bleeding.
Main Methods:
- Mice were pre-treated with compound C or a vehicle control.
- Mesenteric arteriolar thrombosis and tail bleeding times were assessed.
- Platelet aggregation, clot retraction, integrin activation, and spreading were analyzed in vitro.
- Phosphorylation of myosin light chain (MLC) and MYPT1, and RhoA-GTP levels in platelets were measured.
Main Results:
- Compound C significantly reduced mesenteric arteriolar thrombosis but did not alter tail bleeding time.
- Compound C inhibited thrombin- and collagen-induced platelet aggregation, clot retraction, and integrin activation.
- Platelet spreading was impaired by compound C, but phosphatidylserine exposure remained unaffected.
- Compound C decreased thrombin-induced MLC and MYPT1 phosphorylation and reduced RhoA-GTP levels in platelets.
Conclusions:
- AMPK inhibition via compound C demonstrates potential as an antithrombotic strategy.
- The antithrombotic effects are partly mediated by modulating RhoA-MYPT1-MLC signaling pathways involved in cytoskeletal changes.
- Targeting AMPK may offer a way to develop antithrombotic drugs that spare hemostasis.
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