Effects of GYKI 52466 on Early Vasospasm in Rats

Abstract

Objective: The pathogenesis and treatment of cerebral vasospasm (CV) after subarachnoid hemorrhage (SAH) remains a matter of discussion. The authors investigated the efficacy of GYKI 52466, a 2,3-benzodiazepine that is a selective and potent α-amino-3-hydroxy-5-methylisoxazole-4-proprionic acid (AMPA) receptor antagonist, in a rat femoral artery vasospasm model.

Methods: Twenty-seven Wistar albino rats were used in this study. The animals were divided into 3 groups of 9 animals each: sham-operated control (group 1), vasospasm group (group 2), and vasospasm-plus-treatment group (group 3). In groups 2 and 3, autologous blood (0.1 mL) was applied to a 1-cm segment of the femoral artery, which was then wrapped with a silicone cuff. One minute after blood application, the rats in group 3 received an intraperitoneal injection of 15 mg/kg GYKI 52466 every 12 h for 24 h. Responses to blood application and treatment were evaluated with light and electron microscopy examinations of femoral artery specimens at 72 h.

Results: On light microscope examination, the mean diameters of the arterial lumens in groups 1, 2, and 3 were 514.47±15.3, 317.63±12.1, and 503.91±9.6 μm, respectively. At 24 h, the mean arterial wall thickness in group 1 was 77.69±4.2 μm. This mean thickness in group 2 increased to 164.82±9.1 μm. After GYKI treatment in group 3, the mean arterial wall thickness measured 95.37±5.3 μm. In group 2 rats, electron microscopy demonstrated various changes including marked luminal narrowing and increased wall thickness in the femoral arterial wall. The most striking finding were the degenerative changes in the endothelium, which presented as a corrugated appearance of the internal elastic lamina. Rats in group 3 had endothelia that were slightly constricted and smooth muscle cells that were relaxed; changes in the vessel wall and internal elastic lamina were less prominent in these rats than in the rats of group 2.

Conclusions: The results of the present study suggest that GYKI 52466 inhibited AMPA receptors and induced relaxation of smooth muscle cells in the wall of the femoral artery in a rat model. This substance may be a protective and therapeutic agent in the treatment of cerebral vasospasm.

Zusammenfassung

Ziel: Die Pathogenese und die Behandlung des zerebralen Vasospasmus (CV) nach der Subarachnoidalblutung (SAB) bleibt Gegenstand der Diskussion. Die Autoren untersuchten die Wirksamkeit von GYKI 52466, ein 2,3-Benzodiazepin, welches ein selektiver und potenter α-Amino-3-Hydroxy-5-Methylisoxazol-4-Proprionsäure (AMPA) Rezeptorantagonist ist, im Rattenmodell des Vasospasmus der Femoralarterie.

Methoden: 27 Wistar Albinoratten wurden in dieser Studie eingesetzt. Die Tiere wurden in 3 Gruppen von jeweils 9 Tieren eingeteilt: Kontroll-operierte Tiere (Gruppe 1), Vasospasmus-Gruppe (Gruppe 2) und Vasospasmus-Behandlungsgruppe (Gruppe 3). In den Gruppen 2 und 3 wurde autologes Blut (0,1 ml) auf ein 1 cm-Segment der Femoralarterie aufgebracht, welche anschließend in eine Silikonhülse eingewickelt wurde. Eine Minute nach der Blutanwendung erhielten die Ratten der Gruppe 3 eine intraperitoneale Injektion von 15 mg/kg GYKI 52466 alle 12 Stunden für 24 Stunden. Die Reaktionen auf die Blutanwendung und die Behandlung wurden mit licht- und elektronenmikroskopischen Untersuchungen des Femoralarterien-Segments nach 72 Stunden evaluiert.

Ergebnisse: Bei lichtmikroskopischer Untersuchung betrugen die mittleren Durchmesser des Arterienlumens in den Gruppen 1, 2 und 3 jeweils 514,47±15,3, 317,63±12,1 und 503,91±9,6 μm. Nach 24 Stunden betrug die mittlere Wandstärke in Gruppe 1 77,69±4,2 μm. Die mittlere Wandstärke in Gruppe 2 stieg an auf 164,82±9,1 μm. Nach der GYKI-Behandlung in Gruppe 3 betrug die mittlere Wandstärke 95,37±5,3 μm. Bei Ratten der Gruppe 2 ergab die elektronenmikroskopische Untersuchung verschiedene Ver-änderungen, einschließlich deutlicher Lumeneinengung und verstärkter Wanddicke in der Femoralarterienwand. Am bemerkenswertesten waren die degenerativen Veränderungen im Endothel, die sich als wellenförmige Erscheinungen der Lamina elastica interna darstellten. Ratten der Gruppe 3 hatten Endothelien, die leicht zusammengezogen waren und glatte Muskelzellen, die entspannt erschienen. Veränderungen der Gefäßwand und der Lamina elastica interna waren weniger deutlich in diesen Ratten im Vergleich zu den Ratten der Gruppe 2 (Vasospasmus-Gruppe).

Schlussfolgerungen: Diese Ergebnisse zeigen, dass GYKI 52466 die AMPA Rezeptoren hemmt und eine Relaxation der glatten Muskelzellen in der Wand der Femoralarterie im Rattenmodell erzeugt. Diese Substanz könnte ein protektiver oder therapeutischer Wirkstoff bei zerebralem Vasospasmus sein.

Key words

α-amino-3-hydroxy-5-methylisoxazole-4-proprionic acid receptor - cerebral vasospasm - femoral artery - 2,3-benzodiazepine - subarachnoid hemorrhage

Schlüsselwörter

α-Amino-3-Hydroxy-5-Methylisoxazol-4-Proprionsäure-Rezeptor - zerebraler Vasospasmus - Arteria femoralis - 2,3-Benzodiazepin - Subarachnoidalblutung

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