Saikosaponin-d improved the stemness of mouse neural stem cells and increased their thermotolerance potential.

Purpose: To investigate the effect of saikosaponin-d (Ssd) on proliferation, differentiation, and stemness of neural stem cells (NSCs), and to observe whether Ssd has a protective effect on NSCs at medium-high and high temperature.

Materials And Methods: NSCs were extracted from 15-day fetal mice. After subculture, Ssd treatment was performed. Cell cycle and apoptosis rate were detected by flow cytometry. Western Blot and immunofluorescence assay were used to detect the expression and spatial distribution of Nestin, NSE, GFAP, Oct4, and SOX2. Cell growth morphology was observed under a microscope; the concentration of extracellular lactate dehydrogenase (LDH) was determined by ELISA.

Results: Compared with the control group, the proportion of NSCs in the G0/G1 phase increased in the Ssd treatment group; on the contrary, the proportion in the G2/M phase significantly decreased. Microscopically, our results also suggested the sphere-formation rate increased significantly. Besides, the percentage of dead cells in the Ssd group at 38.5, 40°C were reduced, and the level of LDH release was dropped.

Conclusion: Ssd improved the stemness of NSCs, inhibited their differentiation into neural cells, and reduced cell damage under high temperature. Therefore, we speculate that Ssd can improve the thermotolerance of NSCs and protect the nervous system of children with fever.