• Citrullus colocynthis extract could improve the pathological state of Parkinson's disease in MPP + cell model and MPTP mouse model. • Bioinformatics analysis of the functional network of citrullus colocynthis revealed autophagy might involve in the anti-PD effect of citrullus colocynthis . • Citrullus colocynthis extract induced autophagy associated cell death via inhibited ROS-AMPK-ULK1 pathway in MPP + cell model. Citrullus colocynthis (CC), an ethnic medicine, distributed in Xinjiang Uygur Autonomous Region of China, is a traditional remedy for rheumatic arthritis or diseases caused by abnormal black biliary. Here, we investigated the therapeutic effect of CC on Parkinson's disease (PD) and its protective mechanisms. Firstly, the effects of CC on MPTP-induced PD mouse model were evaluated by a series of behavioral tests. Significant improvement of bradykinesia and memory reduction and reduction of climbing time were observed. Meanwhile, improvement of tyrosine hydroxylase (TH) and α-synuclein was also detected by immunohistochemical and western blot analysis. Additionally, it was also found that CC could relieve the oxidative stress in PD mouse brain tissues by decreasing activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and increasing activity of malondialdehyde (MDA). Moreover, possible protective mechanisms were analyzed by a set of combined bioinformatics methods. Subsequently, the mechanism was verified using immunofluorescence, flow cytometry and western blot. Results showed that CC could reduce oxidative damage by inhibiting the autophagy via ROS-AMPK-ULK1 signaling pathway in MPP + -induced SH-SY5Y cells. In conclusion, the present study validated that CC has a promising neuroprotective effect against PD, which can reduce oxidative stress and inhibiting autophagic cell death.