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Schwann cells play an important role in peripheral nerve injury, regeneration and repair.To determine whether Fasudil or RNAi-mediated RhoA gene silencing can optimize Schwann cells culture.Schwann cells were divided into 5 groups: control group; 5 μmol/L Fasudil group; 10 μmol/L Fasudil group; 15 μmol/L Fasudil group; 20 μmol/L Fasudil group; RhoA gene silencing group. Three days later, RT-PCR and Western blot were used to assess the expression of RhoA mRNA and RhoA protein. Cellular proliferation was determined by the cell growth curve (5 days) and MTT assay. The cell cycle was analyzed by flow cytometry.There was no difference between 20 μmol/L and 15 μmol/L Fasudil groups (P > 0.05). Fasudil and RNAi-mediated RhoA gene silencing can promote the proliferation of Schwann cells, and 15 μmol/L Fasudil and RhoA gene silencing have the best effect.