Alfaxalone activates Human Pregnane-X Receptors with greater efficacy than Allopregnanolone: an in-vitro study with implications for neuroprotection during anesthesia

BackgroundAlfaxalone is a fast acting intravenous anesthetic with high therapeutic index. It is an analogue of the naturally-occurring neurosteroid, allopregnanolone which has been implicated in causing neuroprotection, neurogenesis and preservation of cognition, through activation of pregnane X receptors in the central nervous system. This study investigated whether alfaxalone can activate human pregnane X receptors (h-PXR) as effectively as allopregnanolone.MethodsAllopregnanolone and alfaxalone were dissolved in dimethyl sulfoxide to make allopregnanolone and alfaxalone treatment solutions (serial 3-fold dilution concentration range, 50,000 – 206 nM). Activation of h-PXR by these ligand solutions compared with vehicle control was measured by an in-vitro method using human embryonic kidney cells (HEK293) expressing h-PXR hybridised and linked to the firefly luciferase gene. Ligand binding with and activation of h-PXR in those cells caused downstream changes in luciferase activity and light emission. That activity was measured as relative light units using a plate-reading luminometer, thus quantifying the changes in h-PXR activity caused by the ligand applied to the HEK293 cells. Ligand log concentration response curves were constructed to compare efficacy and potency of allopregnanolone and alfaxalone.ResultsAllopregnanolone and alfaxalone both activated the h-PXR to cause dose-related light emission by the linked firefly luciferase. Control solutions (0.1% dimethyl sulfoxide) produced low level light emissions. Equimolar concentrations of alfaxalone were more efficacious in activation of h-PXR: 50,000 nM, p = 0.0019; 16,700 nM, p = 0.0472; 5,600 nM, p = 0.0031 [Brown-Forsythe and Welch ANOVA].ConclusionsAlfaxalone activates human-pregnane X receptors with greater efficacy compared with the endogenous ligand allopregnanolone. These results suggest that alfaxalone sedation and anesthesia may be accompanied by beneficial effects normally caused by the physiological effects of allopregnanolone, namely neuroprotection, neurogenesis, and preservation of cognition.

Meclizine ameliorates memory deficits in streptozotocin-induced experimental dementia in mice: role of nuclear pregnane X receptors

Pregnane X receptors (PXRs) regulate the expression of ATP-binding cassette proteins transporters and organic anion transporting polypeptides responsible for influx/efflux of xenobiotics across the brain. Ligand activation of PXR augments the expression of P-gp and promotes amyloid-β clearance across the blood–brain barrier. Dementia was induced in mice by intacerebroventricular administration of streptozotocin (STZ) followed by treatment with meclizine, a PXR agonist, and subsequently exposed to the Morris water maze test and biochemical and histopathological analysis to evaluate the effect on cognition. STZ-treated mice exhibited significant enhancement in brain thiobarbituric acid reactive species, interleukin-1β, tumour necrosis factor-α, myeloperoxidase, and acetylcholinestrase activity in addition to diminution in glutathione levels and superoxide dismutase activity in comparison to untreated mice. Administration of meclizine to STZ mice recuperated cognition and biochemical alterations. Concomitant administration of ketoconazole, a PXR antagonist, with meclizine prevented the protective effects. The upshots of our study proclaim that meclizine protects cognitive deficits by virtue of its antioxidant, anticholinesterase, and antiinflammatory properties. Results also signify the potential of PXR in neuroprotective actions of meclizine in dementia.