TDZD-8 pre-treatment in transient middle cerebral artery occlusion

Abstract

There is an unmet clinical need to develop neuroprotective agents for cerebrovascular procedures requiring transient cerebral artery occlusion. This study aims to investigate the effects of a single pre-treatment dose of 4-benzyl-2-methyl-1,2,4-thiadiazolidine-3,5-dione (TDZD-8), a glycogen synthase kinase-3β (GSK-3β) inhibitor, in a transient focal cerebral ischemia model. Twenty-eight male adult Wistar rats were subjected to right middle cerebral artery (MCA) occlusion via intraluminal thread technique for 60 min under continuously cortical perfusion monitoring by laser-Doppler flowmetry. Rats were divided into two groups: control or treatment groups. In the treated group, TDZD-8 (5 mg/kg; intravenously) was administered 10 min before the onset of the MCA ischemia. At 24-h reperfusion, the following parameters were evaluated: neurological deficits, brain infarct volume, ipsilateral hemispheric oedema, neuron specific enolase (NSE) plasma levels, parenchyma histology (H–E staining), Fluoro-Jade positive neurons, p-Akt and total Akt expression by western blot analysis and p-Akt-positive nuclei by immunohistochemistry. Infarct volume (P < 0.001) and neurological deficits severity (P < 0.001) were reduced in TDZD-8 treated group. TDZD-8 attenuated hemispheric oedema (P < 0.001), prevented the NSE plasma level increase (P < 0.001) and diminished the number of degenerated neurons in the infarct area (P < 0.001), as shown by Fluoro-Jade staining. TDZD-8 treated rats showed few signs of perivascular oedema when compared to control group. No variations in total Akt and p-Akt expression were observed; instead immunohistochemistry showed increased p-Akt nucleus translocation in TDZD-8 treated rats (P < 0.05). TDZD-8 is a potential pre-treatment intraoperative drug to prevent neuronal injury induced by transitory artery occlusion during cerebrovascular procedures.