Evaluation of Embryotoxicity of Radix scutellariae   Based on Embryonic Stem Test

Purpose: To determine the potential embryotoxicity of Radix scutellariae

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(RS) extract using an embryonic stem cell test (EST) and to evaluate its effect on the differentiation of mouse embryonic stem (ES) cells.
Methods: All the test samples were obtained by water extraction method. The embryotoxicity of RS was assessed with cytotoxicity assays, namely, embryonic stem (ES) cells (IC₅₀ES) and 3T3 fibroblasts (IC₅₀3T3), as well as cardiac differentiation inhibition assay (ID₅₀ES). The expression of specific genes and proteins was analyzed by quantitative reverse transcription – polymerase chain reaction (RT-PCR) and Western blot.
Results: RS was weakly embryotoxic with IC₅₀ES, IC₅₀3T3 and ID₅₀ES of 0.1524, 0.1061, and 0.4169 mg/ml, respectively. Also RS had discordant effects on the expression of tissue-specific genes and proteins in three germ layers, promoting differentiation of the ectoderm (⋆p < 0.05; ⋆⋆p < 0.01) and endoderm (⋆p < 0.05; ⋆⋆p < 0.01; ⋆⋆⋆p < 0.001), while inhibiting mesoderm differentiation (⋆p < 0.05; ⋆⋆p < 0.01; ⋆⋆⋆p < 0.001). The effect of RS on the embryonic development of the three germ layers was concentration-dependent.
Conclusion: These results indicate that RS possesses weak embryotoxicity. The variability in the effects of RS may be responsible for its weak embryotoxicity.

Keywords
Embryonic stem test; Radix scutellariae; Embryotoxicity; Cardiac differentiation inhibition assay; Ectoderm; Endoderm; Mesoderm