Neuroprotective Effect of IRL-1620, an Endothelin B Receptor Agonist, on a Pediatric Rat Model of Middle Cerebral Artery Occlusion

Enrique G Cifuentes, Mary G Hornick, Suresh Havalad, Ramona L Donovan, Anil Gulati, Enrique G Cifuentes, Mary G Hornick, Suresh Havalad, Ramona L Donovan, Anil Gulati

Abstract

Objective: The purpose of this study was to determine the potential neuroprotective effect of endothelin B (ETB) receptor agonist IRL-1620 treatment in a pediatric model of ischemic stroke. Design: A prospective, animal model study. Setting: An experimental laboratory. Subjects: Three-month-old male Wistar Han rats. Interventions: The rats underwent permanent middle cerebral artery occlusion (MCAO). At 2, 4, and 6 h post MCAO, they were treated with saline, IRL-1620 (5 μg/kg, IV), and/or ETB antagonist BQ788 (1 mg/kg, IV). Measurements and Main Results: The rats were evaluated over the course of 7 days for neurological and motor deficit, cerebral blood flow (CBF), and infarct volume. Young rats treated with IRL-1620 following MCAO improved significantly in neurological and motor assessments as compared to the vehicle-treated group, as measured by neurological score (P = 0.00188), grip test (P < 0.0001), and foot-fault error (P = 0.0075). CBF in the infarcted hemisphere decreased by 45-50% in all groups immediately following MCAO. After 7 days, CBF in the infarcted hemisphere of the IRL-1620 group increased significantly (P = 0.0007) when compared to the vehicle-treated group (+2.3 ± 23.3 vs. -45.4 ± 10.2%). Additionally, infarct volume was significantly reduced in IRL-1620-treated rats as compared to vehicle-treated rats (P = 0.0035, 41.4 ± 35.4 vs. 115.4 ± 40.9 mm3). Treatment with BQ788 blocked the effects of IRL-1620. Conclusions: IRL-1620 significantly reduced neurological and motor deficit as well as infarct volume while increasing CBF in a pediatric rat model of cerebral ischemia. These results indicate that selective ETB receptor stimulation may provide a novel therapeutic strategy in the treatment of pediatric ischemic stroke as has been demonstrated in adult ischemic stroke.

Keywords: cerebral blood flow; cerebral infarction; endothelin B receptor agonists; ischemic penumbra; reperfusion; stroke.

Figures

Figure 1
Figure 1
Percent change in cerebral blood flow (CBF). Treatment with IRL-1620 lessens the reduction in CBF after middle cerebral artery occlusion (MCAO). This effect was blocked by treatment with ETB antagonist BQ788. Groups I-III: n = 6, Group IV: n = 5, Group V: n = 4. Values are presented as mean ± SD. P < 0.05: @, compared to vehicle group; #, compared to IRL-1620 group.
Figure 2
Figure 2
Pericam PSI Laser Speckle Contrast Analysis (LASCA) imaging of cerebral blood flow. Top: Group II (vehicle), left to right: brightfield image, baseline, 1 h post MCAO, 7 days post MCAO. Bottom: Group III (IRL-1620), left to right: brightfield image, baseline, 1 h post MCAO, 7 days post MCAO.
Figure 3
Figure 3
Effect of IRL-1620 and BQ788 on infarct volume (mm3). Groups I-III: n = 6, Group IV: n = 5, Group V: n = 4. Values are presented as mean ± SD. P < 0.05: @, compared to vehicle group; #, compared to IRL-1620 group.
Figure 4
Figure 4
Representative images of infarct. Two-mm coronal sections of brains were stained with 2% 2,3,5-triphenyltetrazolium chloride (TTC) to visualize the infarct area. Arrows denote area of infarct. Red, normal tissue; white, infarcted tissue.

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