Evaluation of the Effectiveness of Post-Stroke Metformin Treatment Using Permanent Middle Cerebral Artery Occlusion in Rats.

Gintare Zemgulyte,Katryna Pampuscenko,Norio Sakai,Izumi Hide,Daiva Rastenyte,Vilmante Borutaite,Shigeru Tanaka

doi:10.3390/ph14040312

Abstract

Stroke is the second leading cause of death worldwide. Treatment options for ischemic stroke are limited, and the development of new therapeutic agents or combined therapies is imperative. Growing evidence suggests that metformin treatment, due to its anti-inflammatory action, exerts a neuroprotective effect against ischemia/reperfusion-induced brain damage. Experimental assessment has typically been performed in models of cerebral transient ischemia followed by long-term reperfusion. The aim of this study was to evaluate the neuroprotective effect of metformin treatment after permanent middle cerebral artery occlusion (pMCAO) without reperfusion in rats. Neurological deficits were assessed using the Longa scale, which offers a graded scale on body movement following pMCAO. Both infarct size and brain oedema area were measured by staining with 2,3,5-triphenyltetrazolium chloride. The number of neurons and total and activated microglia, as well as interleukin 10 (IL-10) production, in brain sections were evaluated by immunohistochemical staining. Our results show that metformin treatment improves the neurological state and reduces infarct size after 120 h of pMCAO. Metformin also prevents neuronal loss in the ischemic cortex but not in the striatum after 48 h of pMCAO. Moreover, post-stroke treatment with metformin significantly decreases the number of total and activated microglia at 48 h. The anti-inflammatory effect of metformin is associated with increased IL-10 production at 48 h after pMCAO. The results of the present study suggest that post-stroke treatment with metformin exerts anti-inflammatory and neuroprotective effects in a pMCAO model.

Highlights

Stroke is the second leading cause of death worldwide [1]
Infarct size and area of brain oedema at 48 h and 120 h after permanent middle cerebral artery occlusion (pMCAO) were measured by TTC staining
At 120 h, infarct size in metformin-treated rats decreased by 55% (p = 0.008) compared to the pMCAO group with metformin at 48 h and was reduced by 38% (p = 0.016) compared with the pMCAO

Summary

Introduction

Stroke is the second leading cause of death worldwide [1]. Reperfusion therapy, including intravenous thrombolysis with tissue plasminogen activator and mechanical thrombectomy, is the only approved treatment for acute ischemic stroke [2]. Since the time from stroke onset is a key determinant of patient selection for reperfusion therapy [3], only a portion of ischemic stroke patients are eligible for it [4]. Over the last three decades, various pharmacologic agents and non-pharmacological therapies have been evaluated to protect the brain from ischemic injury [5]. The development of new neuroprotective agents or combined therapies is imperative [10]

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