This work aims to describe the purification and characterization of acidic-hydrolyzed corn silk polysaccharides (AH-CSP) and evaluate their protection on the HO-injured intestinalepithelial cells (IEC-6). Two fractions named AHP-1 and AHP-2 were obtained from AH-CSP, and physicochemical properties of them were investigated by gel permeation chromatography (GPC), gas chromatography (GC), nuclear magnetic resonance (NMR), fourier transform infrared (FT-IR) spectroscopy, scanning electronic microscopy (SEM), and Congo red test. Results showed that AHP-1 (2.80 × 10 Da) and AHP-2 (1.25 × 10 Da) were consisted of xylose, mannose, galactose, rhamnose, arabinose, and glucose. AHP-1 and AHP-2 had strong scavenging activities on 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-Azobis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS), and OH· free radicals. Furthermore, pretreatment with AHP-2 could protect the HO-injured IEC-6 cells by effectively scavenging the overproduced reactive oxygen species (ROS) and regulating of Kelch-like ECH-associated protein 1(Keap1)/ nuclear factor erythroid 2-related factor 2 (Nrf2) signal pathway.