Oleanolic acid significantly inhibited neurosphere formation in a dose-dependent manner, and achieved a maximum effect at 10 nM. OA also reduced the 5-ethynyl-2’-deoxyuridine (EdU) incorporation into NSCs, indicating an inhibition on proliferation of NSCs. Next, in western blotting analysis, the protein expression of neuron-specific marker tubulin-βⅢ (TuJ1) and Mash1 was increased, while the astrocyte-specific marker glial fibrillary acidic protein (GFAP) decreased. In immunofluorescence analysis, OA significantly elevated the percentage of TuJ1-positive cells and reduced GFAP-positive cells. Using DNA microarrays, 183 genes were found to be differentially regulated by OA. Neural stem cells derived form mouse embrynic brains were treated with Oleanolic acid or not. Each group had 4 biological relicates each from a mouse.
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