Objective: To investigate the molecular targets of Modified GanDou decoction (MGDD) treatment in Cyt C/Caspase signaling pathway in neurons of TX neonatal mice

and explore its corresponding mechanism on regulating Cyt C/Caspase signaling pathway. Method: neurons of TX neonatal mice were obtained by using primary culture method

and were divided into normal group (normal DL neonatal mice neurons

cultured by complete medium)

model group (TX neonatal mice neurons

cultured by 10% blank rabbit serum)

MGDD group (TX neonatal mice neurons

continuously cultured by rabbit serum containing 5%

10%

15% and 20% MGDD)

and Butylphthalide group (TX neonatal mice neurons

cultured by rabbit serum containing 10% Butylphthalide). Atomic absorption method was used to detect the effect of different concentrations of MGDD rabbit serum on microelement in the neurons of TX neonatal mice after 24 h treatment flow cytometry was used to analyze the expression of reactive oxygen (ROS) in the neurons of TX neonatal mice after treatment with rabbit serum containing MGDD; Western blot was used to detect the protein expression levels of Cyt C

Caspase-9 and Caspase-3 in the neurons of TX neonatal mice after treatment with rabbit serum containing MGDD. Result: As compared with the model group

MGDD could reduce the contents of copper and iron in the neurons of TX neonatal mice

and increase the content of zinc in a dose-dependent manner (P<0.01). Flow cytometry results showed that MGDD can significantly reduce the release of ROS as compared with model group (P<0.01). Western blot results indicated that MGDD significantly decreased the protein expression levels of Cyt C

Caspase-9

and Caspase-3 in the neurons of TX neonatal mice as compared with the model group (P<0.01). Conclusion: MGDD could relieve the high copper-induced neurons damage

probably by promoting the copper excretion and inhibiting the expression levels of Cyt C

Caspase-9 and Caspase-3 in TX neonatal mice neurons. MGDD achieves the treatment effect by reducing copper content in brain and regulating Cyt C/Caspase signaling pathway.