Effects of Panax notoginseng saponins on proliferation and differentiation of rat embryonic cortical neural stem cells

J Chin Med Assoc. 2016 May;79(5):256-63. doi: 10.1016/j.jcma.2015.10.011. Epub 2016 Feb 22.

Abstract

Background: We aimed to study the effect of Panax notoginseng saponins (PNS) on the proliferation, differentiation, self-renewal, and expressions of basic fibroblast growth factor (bFGF) and brain-derived neurotrophic factor (BDNF) in rat embryonic neural stem cells (NSCs).

Methods: Cortical stem cells were isolated from rat embryos on Embryonic Day 17 (E17) and identified by nestin expression. Subsequently, primary culture, subculturing, and single cell cloning were performed on the cells. After the first cell passage (P1), the cells were resuspended and divided into a control group and a treatment group. Control cells were cultured in serum-free basal culture medium with B27 and dulbecco's modified eagle medium (DMEM)/F12. The same medium supplemented with PNS (100 μg/mL) was used to culture cells in the treatment group. Both groups were incubated at 37°C in a 5% CO2 incubator. Immunocytochemistry was performed 4 days after incubation.

Results: Primary, P1, and P2 cells in the treatment group formed neurospheres, as did single cell clones of the P1 cells in this group. After being cultured for 4 days, the number of nestin-, proliferating cell nuclear antigen (PCNA)-, Tuj-1-, neurofilament (NF)-, vimentin-, glial fibrillary acidic protein (GFAP)-, bFGF-, and BDNF-positive cells significantly increased in the treatment group in comparison to the control group. All positively stained cells could form clear clusters.

Conclusion: PNS can promote rat embryonic cortical NSC survival, self-renewal, proliferation, and differentiation through neurotrophic factors by autocrine or paracrine signaling.

Keywords: Panax notoginseng saponins; basic fibroblast growth factor; brain-derived neurotrophic factor; cell culture; embryonic rat; neural stem cells.

MeSH terms

  • Animals
  • Brain-Derived Neurotrophic Factor / analysis
  • Cell Differentiation / drug effects
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / drug effects*
  • Glial Fibrillary Acidic Protein / analysis
  • Neural Stem Cells / cytology
  • Neural Stem Cells / drug effects*
  • Panax notoginseng*
  • Proliferating Cell Nuclear Antigen / analysis
  • Rats
  • Rats, Wistar
  • Saponins / pharmacology*
  • Vimentin / analysis

Substances

  • Brain-Derived Neurotrophic Factor
  • Gfap protein, rat
  • Glial Fibrillary Acidic Protein
  • Proliferating Cell Nuclear Antigen
  • Saponins
  • Vimentin