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Effects of clopidogrel on the platelet activation response in horses

Marjory B. BrooksDepartment of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850.

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Thomas J. DiversDepartment of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850.

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Ashlee E. WattsDepartment of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850.

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Sally L. NessDepartment of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850.

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Amelia H. FryeDepartment of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850.

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Tracy StokolDepartment of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850.

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Susan L. FubiniDepartment of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14850.

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Abstract

Objective—To evaluate the platelet activation response before and after treatment with clopidogrel in horses.

Animals—12 healthy adult mares.

Procedures—In a masked study, horses (6/group) were randomly allocated to alternately receive placebo or clopidogrel via nasogastric tube at a loading dose of 4 mg/kg followed by 2 mg/kg every 24 hours. Blood samples were collected before and 72 hours after initiation of treatment for ADP- and collagen-induced light transmission aggregometry; determination of closure time in collagen-ADP cartridges; modified thrombelastography for comparison of maximal amplitudes generated by kaolin, reptilase, and reptilase plus ADP activation; and flow cytometric tests to detect platelet fibrinogen binding, P-selectin expression, and phosphatidylserine externalization before and after ex vivo stimulation with thrombin, convulxin, thrombin with convulxin, and calcium ionophore.

Results—Clopidogrel administration induced a significant decrease in mean aggregation response to 5μM and 10μM ADP stimulation; however, 2 horses had resistance to clopidogrel's inhibitory action. Significant differences after clopidogrel treatment were not found in any other tests of platelet function.

Conclusions and Clinical Relevance—Assays using commercially available reagents were configured to measure different variables of the platelet activation response; however, clopidogrel's platelet inhibitory action was only detected by ADP-induced light transmission aggregometry. Results also suggested that horses, like humans, have interindividual variability in response to clopidogrel that may influence the drug's clinical efficacy as an antiplatelet agent.

Abstract

Objective—To evaluate the platelet activation response before and after treatment with clopidogrel in horses.

Animals—12 healthy adult mares.

Procedures—In a masked study, horses (6/group) were randomly allocated to alternately receive placebo or clopidogrel via nasogastric tube at a loading dose of 4 mg/kg followed by 2 mg/kg every 24 hours. Blood samples were collected before and 72 hours after initiation of treatment for ADP- and collagen-induced light transmission aggregometry; determination of closure time in collagen-ADP cartridges; modified thrombelastography for comparison of maximal amplitudes generated by kaolin, reptilase, and reptilase plus ADP activation; and flow cytometric tests to detect platelet fibrinogen binding, P-selectin expression, and phosphatidylserine externalization before and after ex vivo stimulation with thrombin, convulxin, thrombin with convulxin, and calcium ionophore.

Results—Clopidogrel administration induced a significant decrease in mean aggregation response to 5μM and 10μM ADP stimulation; however, 2 horses had resistance to clopidogrel's inhibitory action. Significant differences after clopidogrel treatment were not found in any other tests of platelet function.

Conclusions and Clinical Relevance—Assays using commercially available reagents were configured to measure different variables of the platelet activation response; however, clopidogrel's platelet inhibitory action was only detected by ADP-induced light transmission aggregometry. Results also suggested that horses, like humans, have interindividual variability in response to clopidogrel that may influence the drug's clinical efficacy as an antiplatelet agent.

Contributor Notes

Dr. Watts’ present address is Department of Large Animal Clinical Sciences, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX 77845.

Supported by the Harry M. Zweig Memorial Fund for Equine Research.

The authors thank Scott Baxendell for technical assistance.

Address correspondence to Dr. Brooks (mbb9@cornell.edu).