The Experimental Research Of Effect Mechanism Of The Macelignan On Alzheimer's Disease

Alzheimer's disease is an irreversible, progressive neurodegenerativedisorder that occurs gradually and results in memory loss and a decline inthinking abilities. Macelignan, extracted from Myristica fragrans Houtt(Myristicaceae), known as pala in Indonesia, has been used traditionally asthe spice and the medicinal purposes. To contribute development of theeffective natural medicine for Alzheimer's disease for the theoretical andexperimental foundation, the curative mechanism of Macelignan onAlzheimer's disease was investigated by means of molecular biology,behavior neuroscience and immuno-histology.The effects of lipopolysaccharide(LPS) and Macelignan on proteinexpression of iNOS, COX-2, p-p38, p38, CREB, NF-kB p65 were measuredby the Western blotting technology in vitro, and the signal transmittingpathway in cell of nervous inflammatory and nervous protective effect ofMacelignan when glutamate-induced neurotoxicity were examined. Then theLPS was infused to the fourth ventricle of rats to prepare the animal modelof the senile dementia and Macelignan (10mg/kg/day) was inoculated to therat stomach after the operation. And the effect of LPS and Macelignan onthe learning and memory was assessed by the atrial-unique matching-to-place task in the Morris water maze and the pathological changes in theprepared animal model were checked by immuno-histochemical method inFischer-344 rats.The results are as follow:1. The treatment of Macelignan did not affect cell viability of the BV2cells by WST-8 cell counting kit.2. Glutamate (5ug/ml) made the BV2 cells swell, intercellular spaceclearer, cell multiplication slower and fragments around the cell increased.And the cell group treated with maceligna and glutamate grew flourishing,had compact intercellular structures, and was similar with the control groupin morphology.3. The Western Blotting examination revealed that Macelignan couldinhibit the protein expression of iNOS, COX-2, p-p38, p38 and CREBinduced by LPS. There were significant differences between LPS group and the control group, between Macelignan group and LPS group.4. There was no significant difference of the NF-kB p65 expressionamong the groups in Western blot examination by LPS and Macelignan.5. The animal neuroethology examination revealed that Macelignancould inhibit the extending of escape latent period induced by LPS. Therewere significant differences between LPS group and the control group(P＜0.01), and between Macelignan group and LPS group (P＜0.05) from thethird cycle. Probe examination revealed that there was difference betweenMacelignan group and LPS group in swimming time at the fourth quadrantwith hided platform (P＜0.05).6. The immunohistochemistry examination revealed that Macelignancould inhibit the microglia activated by LPS and stained by OX-6.The conclusions are as follow:1. Macelignan could inhibit damnification of the spatial learning andmemory function induced and the inhibit cytotoxicity of glutamate and byLPS, so has its neuroprotective effect.2. Macelignan could inhibit the activation of microglia induced ahd theexpression of COX-2 induced by LPS in BV2 cell and the by LPS, so has itsanti-inflammatory effect on neurodegenerative disorder.3. Macelignan could inhibit the expression of iNOS induced by LPS, sohas its good anti-oxidation effect.4. Macelignan could inhibit the expression of p-p38 and CREB inducedby LPS. LPS and Macelignan modulate the production of inflammatorymedium by means of effect to MAPK signal pathway.5. Macelignan could not modulate the production of inflammatorymedium by means of effect to NF-kB signal pathway.6. Macelignan could act to microglia as the anti-iflammatory, theanti-oxidation and neuroprotection, so is the exploring potential naturalmedicine.