Prasugrel, a Platelet P2Y ₁₂ Receptor Antagonist, Improves Abnormal Gait in a Novel Murine Model of Thrombotic Hindlimb Ischemia

Abstract

BackgroundThe efficacy of P2Y12 inhibition for the prevention of cardiovascular events in patients with peripheral arterial disease (PAD) has been established. However, the therapeutic effects on ischemic limb complications are less clear. Accordingly, we aimed to develop a novel murine model of thrombotic hindlimb ischemia to reflect that found in patients with PAD exhibiting ischemic limb symptoms. We further investigated the effects of P2Y12 deficiency and P2Y12 inhibition by prasugrel in this model.Methods and ResultsThrombus formation induced by application of ferric chloride to the femoral artery resulted in a significant reduction in blood flow in the injured limb. In gait analysis using the CatWalk system, moderate difficulties in grounding and weight bearing of the ischemic limb, including reduction of maximum contact area and stance phase duration and increasing in swing phase duration in the ischemic limb, were observed in this model. Blood flow reduction and gait abnormalities gradually recovered over 21 days to levels present before arterial injury. Compared to wild‐type (WT) mice, significant increases in blood flow and improvement in gait were observed in P2Y12‐deficient mice. In addition, daily oral administration of prasugrel (3 mg/kg per day) to WT mice resulted in significant inhibition of blood flow reduction and gait abnormalities to levels found in P2Y12 deficient mice.ConclusionsAcute femoral artery thrombosis resulted in hindlimb ischemia and moderate gait abnormalities in mice. In addition, the present study suggests a possible role of P2Y12 in the complications with thrombotic limb ischemia.

Induction of Diabetes Abolishes the Antithrombotic Effect of Clopidogrel in Apolipoprotein E–Deficient Mice

“…These models of atherothrombosis in mice have been in use for several years. 30 31 32 33 34 35 All studies involving mice were reported in accordance with the ARRIVE guidelines. 37 38…”

“…30 31 32 33 The FeCl 3 -induced thrombosis model is widely used as a standard method by thrombosis researchers. 34 35 In this article, we used this thrombosis model to study the antithrombotic effects of clopidogrel in STZ-treated apoE-deficient mice (diabetic apoE-deficient mice) compared with its antithrombotic effects in wild-type (WT) and nondiabetic apoE-deficient mice (apoE-deficient mice). Further, we determined the antiplatelet effects of clopidogrel in these three groups of mice.…”

Intravascular hemolysis triggers ADP-mediated generation of platelet-rich thrombi in precapillary pulmonary arterioles

“…Some mice were given intravascular heparin (300 U/kg) or eptifibatide (10 mg/kg) 15 minutes before initiation of pulmonary thrombosis (66,67). Prasugrel was suspended in a 5% solution of gum arabic (68) and administered once a day for 2 days and, additionally, approximately 4 hours before qFILM to each mouse by PO with a volume of 10 mL/kg. Physiological saline was intravascularly administered to mice as a vehicle in control experiments.…”