Role of Pomegranate Juice and Atorvastatin in Ameliorating Spinal Neurotoxicity of Wistar Rats Maternally Fed on Hypercholesterolemic Diet

Abstract

Background: The contribution of high dietary cholesterol to encephalomyelitis and retinal diseases give more attention to overcome its neurotoxicity. These led physician and scientists of using anti-lepidemic statin drugs, however their long-term treatment may develop complication. Today many views recommended supplements of natural products especially fruit to get demand of needed antioxidant to scavenge bad free radicals liberated from inflammation. Pomegranate juice supplementation is used in the present study in combination with atorvastatin drug hoping to give synergistic effects and improve neurotoxicity. Material and methods: Eighty pregnant Wistar albino rats weighing 180 g to 200 g body weight were used. They were arranged into eight groups (n=8) such as control, atorvastatin (1 mg/kg), pomegranate (0.5 ml 50%/ rat), atorvastatin and pomegranate, hyper ccholesterolemic group (3% cholesterol diet for 6 weeks before onset of gestation), hypercholesterolemia and atorvastatin and/or pomegranate. All treatments were maintained throughout gestation and lactation period till 2 and 3 week-old. Offspring were sacrificed by diethyl ether and their cervical spinal cord were dissected and separated. The spinal cord was subjected for histological and transmission electron microscopical investigations, biochemical assays of neurotransmitters (dopamine, serotonin and γ-Aminobutyric acid), vascular endothelial growth factors, 8-hydroxydeoxyguanosine, caspase 3 and caspase 7 as well as comet assay). Results: The present findings revealed massive cell death and necrosis within ependymal canal as well as pyknosis and edematous lesions of neuronal cells in offspring of hypercholesterolemic mothers. At ultrastructural level, there is a detected demyelination and vacuolar degeneration of myelinated axons. Many of sensory and motor neuronal cells exhibited compacted chromatin materials of their nuclei. These were associated with depletion of assayed neurotransmitters (DA and 5-HT), overexpression of vascular endothelial growth factors, caspases 3 and 7 and 8-hydroxydeoxyguanosine. Stretched DNA damage (comet assay) with obvious head and tail region was detected in neuronal cells of offspring of hypercholesterolemic mothers and decreased post-pomegranatesupplementation. Atorvastatin and pomegranate supplementation exhibited apparent improvement comparing with single atorvastatin or pomegranate manifesting highly synergistic effects. Conclusion: Feeding pregnant on hypercholesterolemic diet induced spinal cord injury of their offspring and these can be improved by pomegranate juice supplementation in combination with atorvastatin- treatment.