Objective To investigate the protective effects of ethanol extract of Chuangxiong tea on the 2,2'-azobis(2-methylpropionamidine) dihydrochloride (AAPH) (1 mmol/L) on oxidative stress injury of proximal renal tubule epithelial cells of pigs. Methods LLC-PK1 cells were co-cultured with ethanol extract of Chuanxiong tea (labeled CXTEE) with different mass concentrations ranging from 10 to 100 μg/mL for 24 h, and then placed in Dulbecco's modified eagle medium (DMEM) containing AAPH for 4 h to establish the cell damage model. The cell survival rate was measured by tetramethyl azazole blue (MTT) method. The content of malondialdehyde (MDA) and the level of total reactive oxygen species (ROS) were determined by thiobarbituric acid colorimetry and 2',7'-dihydrodichlorofluorescein yellow diacetate (DCFH-DA) probe technique, respectively. In addition, the activity of several antioxidant enzymes, including catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione S-transferase (GST), γ-glutamylcysteine synthetase (γ-GCs), and the concentration of glutathione (GSH), were also evaluated using specific kits. Results The results showed that the cells pretreated with CXTEE showed a higher survival rate, and the total ROS level and MDA production in the cells were significantly reduced. At the same time, the extract also enhanced the activity of antioxidant enzymes (such as CAT, SOD, GSH-Px and GST) in the damaged cells, promoted the activity of γ-GCS and increased the content of GSH. Further experiments showed that CXTEE could also up-regulate the mRNA expression levels of SOD, GSH-Px and CAT in oxidative damage LLC-PK1 cells. Conclusion The ethanol extract of Chuangxiong tea may reduce the levels of MDA and ROS by strengthening the antioxidant defense system of cells, thus alleviating the oxidative stress damage caused by AAPH to LLC-PK1 cells. This study can provide a basis for further application and development of Chuangxiong tea.