Objective To investigate the effect of theca cells on granulosa cells injured by chemotherapy and the intervention effect of serum containing Zuogui pill ( ZGW). Methods Serum containing Zuogui pill components was prepared. Granulosa and theca cells were cultured and divided into 8 groups, including blank control group, coculture group, ZGW serum group, ZGW serum+coculture group, model group, model+coculture group, model+ZGW serum group and model+ZGW serum+coculture group. The expression levels of Bax, Caspase-3, Beclin 1, p62 and LC3B were detected by qRT-PCR and Western blot. Results Beclin-1, LC3B, Bax and Caspase-3 were highly expressed in the model group at the transcriptional and translational levels, which were downregulated after adding 10% serum containing Zuogui pill components or co-culturing with theca cells. The gene expression of p62 was lower in the model group than that in the control group, and was upregulated by adding 10% serum containing Zuogui pill components or co-culturing with theca cells. When the experimental group was co-cultured with theca cells, the expression of these genes changed more substantially after the addition of 10% serum containing Zuogui pill components. Conclusions Theca cells alleviated the chemotherapy-induced injury of granulosa cells in vitro and demonstrated a synergistic effect with serum containing Zuogui pill components. These effects were related to the inhibition of autophagy and reduction in apoptosis of granulosa cells.