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The purpose of this study was to investigate the effect of nardosinone(Nar) on the proliferation,migration and differentiation of mouse neural stem cells(NSC).The NSC were isolated from mice embryos at gestational day E14 and cultured using the neurosphere culture system.Nar or vehicle was added to the culture media at the start of culture.Cell viability and proliferation were measured with MTT and BrdU incorporation assays respectively.Cell cycle was analyzed by flow cytometric cell sorting.The NSC migration and differentiation were monitored with immune-fluorescence techniques.ERK/CREB signaling pathway was detected with Western blotting analysis.The results demonstrate that after 3 d incubation with the cells,Nar 50 μmol·L-1 increased the viability,BrdU incorporation,and the proportion of cells in S and G2 phase(P<0.05).Exposure to Nar for 24 h promoted cell migration in a dose-dependent manner.Quantitative analyses revealed Nar 50 μmol·L-1 and 25 μmol·L-1 increased the proportion of neurons and oligodendrocytes differentiated from NSC after 7 d and 14 d(P<0.05).Nar,co-incubation with the NSC in proliferation media for 3 d,increased the level of phosphorylated ERK1/2(p-ERK1/2) and phosphorylated CREB(p-CREB) in NSC(P<0.05) without altering the expression of ERK1/2 and CREB.Nar 50 μmol·L-1 and 25 μmol·L-1 also increased the level of p-ERK1/2 and p-CREB in NSC after 7 d co-incubation in differentiation media,however,only increased p-ERK1/2 was observed after 14 d treatment.Together,these findings suggest that Nar promotes the proliferation and migration of NSC,and influence the preference of NSC differentiation.These effects may relate to the action on ERK/CREB signaling pathway.
The purpose of this study was to investigate the effect of nardosinone (Nar) on the proliferation, migration and differentiation of mouse neural stem cells (NSC). The NSCs were isolated from mice embryos at gestational day E14 and cultured using the neurosphere culture system. Nar or vehicle was added to the culture media at the start of culture. Cell viability and proliferation were measured with MTT and BrdU incorporation assays respectively. Cell cycle was analyzed by flow cytometric cell sorting. NSC migration and differentiation were monitored with immune-fluorescence techniques. ERK / CREB signaling pathway was detected with Western blotting analysis. The results demonstrate that after 3 d incubation with the cells, 50 μιηοΐ ^ L-1 increased viability, BrdU incorporation, and the proportion of cells in S and G2 phases (P <0.05) .Exposure to Nar for 24 h promoted cell migration in a dose-dependent manner. Quantitative analyzes revealed 50 μmol·L-1 and 25 μmol·L-1 increased the propor tion of neurons and oligodendrocytes differentiated from NSCs after 7 d and 14 d (P <0.05) .Nar co-incubation with the NSC in proliferation media for 3 d, increased the level of phosphorylated ERK1 / 2 (p-ERK1 / 2) and phosphorylated CREB (p-CREB) in NSC (P <0.05) without altering the expression of ERK1 / 2 and CREB.Nar 50 μmol·L-1 and 25 μmol·L-1 also increased the level of p-ERK1 / 2 and p-CREB in NSC after 7 d co-incubation in differentiation media, however, only increased p-ERK1 / 2 was observed after 14 d treatment. Together, These findings suggest that Nar promotes the proliferation and migration of NSC, and influence the preference of NSC differentiation. These effects may relate to the action on ERK / CREB signaling pathway.