ABSTRACT
Objective To study the effect and mechanism of senegenin on proliferation and differentiation of neural stem cells (NSCs). Methods The primary cultured NSCs were divided into the high-dose, medium-dose, low-dose group and normal control group (NC). The complete medium containing 10, 20 and 40 μmol/L senegenin was added to senegenin low-, middle-, and high- dose groups, and the NC group was routinely cultured. After 4 days of culture, CCK8 assay was used to detect cell viability, and microscopy was performed and the number of neurospheres was counted. Western blot was used to detect the expression of Nestin, TUJ1, GSK-3β, and p-GSK-3β (Ser9), and immunofluorescence staining was used to visualized Nestin and TUJ1. Results Compared with the control group, the number of NSCs neurospheres (32.78 ± 6.30, 40.93 ± 8.34, 45.37 ± 7.96 vs. 26.48 ± 5.19) and the proliferation (127.50% ± 9.31%, 138.13% ± 6.88%, 151.25% ± 9.38% vs. 100.00% ± 5.63%) in the low-, middle- and high-doses of senegenin group significantly increased (P<0.05 or P<0.01).The expression of TUJ1(2.21 ± 0.14,3.10 ± 0.16,3.30 ± 0.15 vs.1.00 ± 0.00)in the low-,middle- and high-doses of senegenin group significantly increased (P<0.05); and the expression of Nestin (0.36 ± 0.04,0.53 ± 0.05,0.46 ± 0.05 vs.1.00 ± 0.00)significantly decreased(P<0.05).The ration of p-GSK-3β(Ser9)/GSK-3β(2.31 ± 0.17,3.41 ± 0.11,3.59 ± 0.16 vs.1.00 ± 0.00)in the low-,middle-and high-doses of senegenin group significantly increased(P<0.01).The cell number of Nestin+(50.29 ± 3.18,45.28 ± 6.23,38.72 ± 5.31 vs. 75.27 ± 6.03) in the low-, middle- and high-doses of senegenin group significantly decreased (P<0.05 or P<0.01), and the cell number of TUJ1+(32.23 ± 4.36,38.23 ± 6.01,46.23 ± 4.36 vs.20.31 ± 5.23)significantly increased (P<0.01). Conclusions The senegenin may promote the proliferation and differentiation of NSCs through the activation of Wnt pathway.