Preclinical evidence of neuroprotective effects of endocannabinoids and related lipid molecules in ischemic stroke: a systematic review and meta-analysis of animal studies

Abstract

Ischemic stroke is an acute cerebrovascular disorder caused by interruption of cerebral blood supply, frequently resulting in severe neurological deficits. However, effective therapeutic strategies remain limited. Recent studies have demonstrated that endocannabinoids (eCBs) and their related N-acylethanolamines (NAEs) exhibit neuroprotective properties as lipid signaling molecules, offering a promising therapeutic approach for ischemic stroke. This meta-analysis aims to systematically evaluate the efficacy and underlying mechanisms of these lipid mediators in animal models of ischemic stroke, thereby providing novel therapeutic targets for intervention. A systematic search was performed across Web of Science, PubMed, Embase, ScienceDirect, and the Cochrane Library for animal studies investigating eCBs and related lipid molecules in the treatment of ischemic stroke, covering publications from 2000 to 2025. The risk of bias for included studies was assessed using the SYRCLE tool. Data were pooled using the appropriate statistical model, with subgroup analysis conducted to explore sources of heterogeneity. A total of 24 studies were included in this analysis. The results demonstrated that eCBs and related lipid molecules significantly reduced cerebral infarct volume, improved neurological function score (NFS), and preserved blood–brain barrier (BBB) integrity, as evidenced by decreased BBB permeability and alleviated cerebral edema. Additionally, these interventions modulated neuroinflammation by inhibiting the phosphorylation of NF-κB and ERK1/2 proteins, reducing TNF-α, IL-1β, and COX-2 levels, and increasing IκBα levels. They also regulated apoptotic markers, decreasing TUNEL-positive cell counts and the levels of Bax and caspase-3 while upregulating Bcl-2. Subgroup analysis revealed significant therapeutic effects with AEA, N15, OEA, PEA, and Δ3-NAE interventions. Moreover, the results obtained were more robust with isoflurane anesthesia, intraperitoneal injection, and post-occlusion administration. The eCBs and NAEs exert anti-inflammatory and anti-apoptotic effects by modulating NF-κB and MAPK signaling pathways, as well as restoring Bax/Bcl-2 balance, thereby demonstrating neuroprotective potential in animal models of ischemic stroke. Future high-quality studies are warranted to further validate the therapeutic value of these lipid molecules, offering novel avenues to overcome the current limitations in ischemic stroke treatment.