Reduced infarct size and accumulation of microglia in rats treated with WIN 55,212-2 after neonatal stroke

doi:10.1016/j.neuroscience.2012.01.008

Neuroscience

Volume 207, 5 April 2012, Pages 307-315

https://doi.org/10.1016/j.neuroscience.2012.01.008 Get rights and content

Abstract

Cannabinoids have emerged as brain protective agents under neurodegenerative conditions. Many neuroprotective actions of cannabinoids depend on the activation of specific receptors, cannabinoid receptor type 1 (CB1R) and type 2 (CB2R). The aim of the present study was to determine whether the CB2R and CB1R agonist WIN 55,212-2 (WIN) protects neonatal brain against focal cerebral ischemia-reperfusion and whether anti-inflammatory mechanisms play a role in protection. Seven-day-old rats were subjected to 90-min middle cerebral artery occlusion (MCAO), and injured rats were identified by diffusion-weighted MRI during the occlusion. After reperfusion, rats were subcutaneously administered 1 mg/kg of WIN or vehicle twice daily until sacrifice. MCAO led to increased mRNA expression of CB2R (but not CB1R), chemokine receptors (CCR2 and CX3CR1), and cytokines (IL-1β and TNFα), as well as increased protein expression of chemokines MCP-1 and MIP-1α and microglial activation 24 h after MCAO. WIN administration significantly reduced microglial activation at this point and attenuated infarct volume and microglial accumulation and proliferation in the injured cortex 72 h after MCAO. Cumulatively, our results show that the cannabinoid agonist WIN protects against neonatal focal stroke in part due to inhibitory effects on microglia.

Highlights

▶WIN 55,212-2 protects the neonatal brain against focal ischemia. ▶Expression of CB2R, but not of CB1R, is induced after neonatal focal ischemia. ▶WIN 55,212-2 prevents microglia/macrophage accumulation after neonatal stroke. ▶WIN 55,212-2 does not affect expression of chemotactic receptors in microglia. ▶WIN does not affect expression of cytokines/chemokines after neonatal stroke.

Section snippets

Animal model of neonatal stroke

All animal research was approved by the University of California San Francisco Institutional Animal Care and Use Committee and was performed in accordance with the Guide for the Care and Use of Laboratory Animals (US Department of Health and Human Services, Publication Number 85-23, 1985). Sprague–Dawley dams with a dated litter of pups were purchased from Charles River Laboratories (Wilmington, MA, USA). Transient 90 min right middle cerebral artery occlusion (MCAO) was performed in 7-day-old

WIN reduces infarct size 72 h after neonatal stroke

The extent of injury was determined 72 h after 90-min MCAO in P7 rats by stereological evaluation of infarct volume (Cavalieri probe) in Nissl-stained brain serial sections. Rats treated with WIN had a significantly smaller infarct volume than rats that received vehicle (11.04±6.02% vs. 19.37±9.01% of injured hemisphere in MCAO+WIN and MCAO+veh, respectively) (Fig. 1A, B vs. 1C, D; Fig. 1F). The volumes of tissue with abnormal DWI during MCAO, that is, tissue at risk, were similar in both

Discussion

We show for the first time that the mixed CB1R and CB2R agonist WIN significantly reduces brain injury during the reperfusion phase after neonatal focal stroke. We also demonstrate that reduction of injury size produced by WIN is associated with decline in proliferation and accumulation of activated microglia in the injured cortex.

It is well known that expression of CB2R is up-regulated in microglia and peripheral monocytes in response to inflammatory stimuli (Maresz et al., 2005, Mukhopadhyay

Acknowledgments

This work was supported by grants from NIH RO1 NS055915 and NIH ROI NS44025 (Z.S.V.); Spanish Ministry of Science and Innovation SAF2009-08145 (M.A.M.) and SAF2008-03122 (I.L.); and Spanish Ministry of Health RENEVAS RD06/0026/0005 (I.L.).

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Neurodegeneration, Neuroprotection, and Disease-Oriented NeuroscienceResearch PaperReduced infarct size and accumulation of microglia in rats treated with WIN 55,212-2 after neonatal stroke

Abstract

Highlights

Section snippets

Animal model of neonatal stroke

WIN reduces infarct size 72 h after neonatal stroke

Discussion

Acknowledgments

Neurosci Lett

Thromb Res

Brain Res

Mol Cell Neurosci

Cytokines

Cell

Pediatr Neurol

Neuroscience

Mol Cell Neurosci

J Neuroimmunol

J Neuroimmunol

Microvasc Res

Neuroscience

Cannabinoid control of neuroinflammation related to multiple sclerosis

Br J Pharmacol

Macrophages are comprised of resident brain microglia not infiltrating peripheral monocytes acutely after neonatal stroke

J Neurochem

Evolution of brain injury after transient middle cerebral artery occlusion in neonatal rats

Stroke

Microglial cells contribute to endogenous brain defenses after acute neonatal focal stroke

J Neurosci

Characterization of the neuroprotective effect of the cannabinoid agonist WIN-55,212 in an in vitro model of hypoxic-ischemic brain damage in newborn rats

Pediatr Res

The cannabinoid agonist WIN55,212 reduces brain damage in an in vivo model of hypoxic-ischemic encephalopathy in newborn rats

Pediatr Res

The cannabinoid WIN55,212-2 promotes neural repair after neonatal hypoxia-ischemia

Stroke

Minocycline confers early but transient protection in the immature brain following focal cerebral ischemia-reperfusion

J Cereb Blood Flow Metab

Erythropoietin enhances long-term neuroprotection and neurogenesis in neonatal stroke

Dev Neurosci

Neurodegeneration, Neuroprotection, and Disease-Oriented Neuroscience
Research Paper
Reduced infarct size and accumulation of microglia in rats treated with WIN 55,212-2 after neonatal stroke