Neuroprotective effect of 2-hydroxy arachidonic acid in a rat model of transient middle cerebral artery occlusion

Abstract

Stroke modifies the composition of cell membranes by eliciting the breakdown of membrane phospholipids whose products, such as arachidonic acid (AA), are released in the cytosol. The action of enzymes such as cyclooxygenases on AA leads to inflammatory stimuli and increases the cell oxidative stress. We report here the neuroprotective effect of 2-hydroxyarachidonic acid (2OAA), a cyclooxygenase inhibitor derived from AA, as a promising neuroprotective therapy against stroke. The effect of a single dose of 2OAA, administered intragastrically 1h after the ischaemic insult, in a rat model of transient middle cerebral artery occlusion (tMCAO) was tested after 24h of reperfusion. Infarct volume was measured by TTC method to evaluate the neuroprotective effect. Levels of phospholipids and neutral lipids were measured by thin-layer chromatography. The expression of cPLA2 and sPLA2 phospholipases responsible for the cleavage of membrane phospholipids, as well as the expression of antioxidant enzymes, was measured by qPCR. Lipid peroxidation was measured as the concentration of malondialdehyde and 4-hydroxynonenal. The treatment with 2OAA reduced the infarct volume and prevented ischaemia-induced increases in transcription levels of free fatty acid (FFAs), as well as in both phospholipases A2 (cPLA2 and sPLA2). The lipid peroxidation and the transcription levels of antioxidant enzymes induced by ischaemia were also decreased by this treatment. We conclude that 2OAA treatment results in a strong neuroprotective effect that seems to rely on a decrease in PLA2 transcriptional activity. This would reduce their action on the membrane phospholipids reducing reactive oxygen and nitrogen species generated by FFAs. Based on the transcriptional activity of the antioxidant enzymes, we conclude that the treatment prevents oxidative stress rather than promoting the antioxidant response. This article is part of a Special Issue entitled: Membrane Lipid Therapy: Drugs Targeting Biomembranes edited by Pablo V. Escribá.

Post‐ischemic salubrinal administration reduces necroptosis in a rat model of global cerebral ischemia

“…Previous studies allow us to estimate that we could detect changes at p ≤ 0.05 level with a statistical power of ≥ 80% with n ≥ 5 rats/ group (Llorerente et al ; Llorerente et al ; Anuncibay‐Soto et al ; Ugidos et al ; Anuncibay‐Soto et al ). Thus, prior to experimental procedure, the number of animals to be used in each group was established in five.…”

The Implications for Cells of the Lipid Switches Driven by Protein–Membrane Interactions and the Development of Membrane Lipid Therapy

“…Accordingly, 2-hydroxy arachidonic acid (2-OAA) is a rationally designed derivative of AA known to be a competitive inhibitor of COX-1 and COX-2, and thus, it can be used in LPS-treated mice to decrease proinflammatory cytokines in serum (reviewed in [212]). When assessed for the treatment of stroke using S-D rats, 2-OAA treatment in the first hour of reperfusion after the stroke produced a neuroprotection [213]. At the molecular level, 2-OAA decreased phospholipase A2 (PLA2) in the cell membrane with a subsequent decrease in FFA release.…”

“…A decrease in oxidative stress also occurs upon 2-OAA treatment in the first hour of reperfusion after stroke. Thus, the use of rationally designed lipids would seem to be a promising new stroke therapy, with an important advantage that they cross the Blood-Brain Barrier [213].…”

Edad y respuesta a las proteínas mal plegadas en el accidente cerebro vascular. El efecto de la edad = Age and unfolded protein response enhancement in stroke-induced inflammation

“…The Sal treatment included antiinflammatory changes that suggested that its combination with anti-inflammatory agents could result in positive synergic effects against stroke. In this regard, the neuroprotective effect of some anti-inflammatory agents such as meloxicam or 2-OAA has been proved in global and other cerebral brain ischemia models (Llorente et al, 2015;De los Reyes and Céspedes 2014;Ugidos et al, 2017). The development of preferential COX-2 inhibitors such as "coxibs" has resulted in surprisingly different effects, making it necessary to test for each coxib member the risks and benefits in each application (Cairns, 2007).…”

“…In fact, a primary contribution of COX-1 and a delayed contribution of COX-2 in the progression of the damage have been reported in a global cerebral ischemia model (Candelario-Jalil et al, 2003). Thus, the use of NSAIDs with activity against COX-1 and COX-2, which have proved to have a strong neuroprotective effect in tMCAO models, has been claimed (Candelario-Jalil and Fiebich, 2008;Ugidos et al, 2017). Traditional NSAIDs seem to be able to balance the inhibition of the synthesis of platelet tromboxane (anti-trombotic activity) and the synthesis of prostacyclin in endothelial cells (prothrombotic activity).…”