Chinese herbal medicines (CHMs), known for their safety, low toxicity, and minimal impact on the environment, are often used as a natural feed additive in aquaculture to prevent or control diseases of aquatic animals. However, their effect on cell responses of Chinese soft-shelled turtle (Pelodiscus sinensis) remains unexplored. In this study, the extract mixed with four Chinese herbs of Astragalus membranaceus, Lycium barbarum, Rhizoma polygonati and Codonopsis pilosula in proportions (7:8:3:2) was used as a feed additive for 60 days to study their effects on the growth performance and immune response of P. sinensis in greenhouse mode. Results showed that the weight gain rate and the specific growth rate in the CHMs group were 46.63 % and 0.64 %, which were 1.13- and 1.15-fold of that in the control group, respectively. Besides, CHMs could significantly improve the survival rate by 76.94 % after artificial infection of Aeromonas hydrophila. The CHMs group also exhibited significant increases in the activities of total superoxide dismutase (27.76 %), catalase (41.28 %), glutathione peroxidase (14.62 %), and glutathione sulfur transferase (36.75 %) compared to the control group. Conversely, malondialdehyde concentration was reduced to 67.82 % of that in the control group. Moreover, the concentration of lysozyme and complement C3, as well as the activity of acid phosphatase, rose by 16.43 %, 24.30 %, and 27.68 %, respectively. Comparative transcriptome analysis revealed that growth, immunity, and lipid metabolism-related pathways are involved in the response to CHMs. Among these, the two pathways of the cell cycle and the cytokine-cytokine receptor interaction may play important roles in the growth and immunity of P. sinensis. SMOX, STAT3, and HMGCS1 may serve as key genes in CHMs-stimulated immune response in P. sinensis; GADD45B, ARG1, ACVR1C, and BMP7 may be crucial genes in promoting the growth of P. sinensis. This study provides a theoretical and practical basis for the green and efficient culture of P. sinensis.