Isolation and Neuroprotective Effect of[4(3,4-Dihydroxybenzoyloxymethyl)Phenyl-Î-D Glucopyranoside] from Origanummicranthum on In Vitro Glutamate ToxicityModel of Sprague Dawley Rat Cortex Neurons

Glutamate toxicity is a common neurological disorder characterized by an increase in the glutamate (Glu) concentration leads to excitotoxicity which cause cellular damage and death in the brain. Origanum micranthum is an endemic species in Turkey. Origanum species have many polyphenolic compounds. Rich polyphenol containing material is known to be highly neuroprotective effect. Based on this subject, we aimed to evaluate the neuroprotective effect of Origanum micranthum against glutamate-induced toxicity in the primery neuron culture of rat brain cortex. For modeling glutamate toxicity the cells cultures were exposed to 10-5 M glutamate for 20 min. Later, glutamate-induced cell cultures were treated with methanol, chloroform, ethyl acetate and aqueous extracts of aerial parts of Origanum micranthum for 24 h. Cell viability was evaluate by using MTT assay. MTT assay showed that ethyl acetate extract in 10-4 and 10-5 mM concentration have greater neuroprotective effect. [4(3,4-dihydroxybenzoyloxymethyl) phenyl-s- D-glucopyranoside] was isolated majority from ethyl acetate extract of Origanum micranthum by using chromatographic method and evaluated by 1H-NMR (400 MHz), 13C-NMR (100 MHz) and ESI-MS. MTT assay and Anexin V apoptose assay kit show 25, 50 and 100 µgr/ml concentrations of [4(3,4-dihydroxybenzoyloxymethyl) phenyl-s-D-glucopyranoside] exhibited a high neuroprotective activity at microgram concentrations.