Effect of oryzanol, ketone body (acetone) and L-arginine on transient middle cerebral artery (tMCAO) occlusion

Abstract

Aim and Objectives: A stroke is the sudden death of brain cells in a localized area of brain due to inadequate blood flow, leads to hemiparesis, hemisensory deficits, visual field deficits, facial droop, ataxia, aphasia etc. Stroke is second leading cause of death in worldwide. Several report indicates that Oryzanol has free radical scavenging and anti-inflammatory property. Many studies report that ketone bodies have neuroprotective effect on brain because of alternate source of energy. Nitric Oxide may produce neurotoxic or neuroprotective effect, because of produce vasodilation and improve blood flow by reducing by hypoxic injury. This study is aimed to evaluate effect of Oryzanol, ketone bodies and L-Arginine on transient middle cerebral artery occlusion (tMCAO) induced ischemic stroke. Materials and Methods: Wistar rats were used for experiment. Rats are divided into 6 groups: Sham Operated control, tMCAO control, L-Arginine treatment (250mg/kg) in tMCAO, Acetone treatment (10mmol) in tMCAO, Acetone and L-Arginine treatment in tMCAO, and Oryzanol treatment (1μg/μl and not more than 60μl) in tMCAO. Acetone and L-Arginine treatment were started 4days before the hypoxia induction and Oryzanol treatment was started 10days before the Hypoxia induction. Cerebral ischemia was induced by tMCAO using 4-0 intraluminal filament coated with poly-l-lysin. After 24 hours of surgery animals get sacrificed and brain was isolated and infarct volume was measured. Brain homogenate was used for estimation of various biochemical parameters like Caspase-3, LDH, CRP, and antioxidant levels. Expression of apoptotic genes(BAK1 and BAX) and inflammatory gene(NFⱪb) was performed. Immunohistochemistry was performed to assess S100 protein(neurological marker) using rabbit S100 antibodies. Results: Oryzanol, Acetone and L-Arginine showed ameliorative effect on hypoxic injury to the brain due to transient middle cerebral artery occlusion. The drugs showed beneficial effects in brain reperfusion injury and improves the anti-oxidant, anti-inflammatory and anti apoptotic effects. However, combination of acetone and L-arginine did not show advantageous effect over alone acetone or L-arginine treatment.