Abstract:

Objective To incubate rat neural stem cells (NSCs) with selection-specific inhibitor (LY294002) of PI3-K, and explore the effects of PI3-K/AKT signal transduction pathway in NSCs. Methods NSCs derived from E15-16 rats were isolated and cultured. After incubation of NSCs with different concentrations of LY294002 (0 to 40 μmol/L), cell survival (WTS-8 assay), proliferation (BrdU immunohistochemical identification), differentiation (β Tubulin-Ⅲ immunohistochemical identification) and AKP phosphorylated protein (Western blotting) were detected. Results The effects of LY294002 on the survival, proliferation and differentiation of NSCs was concentration-dependent. The survival of the NSCs decreased significantly with a higher concentrations of LY294002 (25, 30, 35, 40 μmol/L)(P<0.05). The numbers of BrdU positive cells with LY294002 of 30, 35 and 40 μmol/L were significantly smaller than that of the control (LY294002 0 μmol/L)(P<0.05). The numbers of β Tubulin-Ⅲ positive cells with LY294002 of 35 and 40 μmol/L were significantly smaller than that of the control (P<0.05). The expression of AKT was blocked by LY294002 in a concentration-dependent manner. Conclusion PI3-K/AKT signal transduction pathway plays a vital role in the survival, proliferation and differentiation of NSCs.

Key words: phosphatidylinositol 3 kinase, protein kinase B, neural stem cells, rat