Impairment of intestinal epithelium barrier is a hallmark of gut pathology. Cell death can compromise barrier function and impair epithelial restitution directly or indirectly in inflammatory bowel disease (IBD). Our previous work demonstrated that glucomannan from Aloe vera gel (AGP) protected mice from DSS-induced colitis, with unclear mechanism of AGP-intestinal barrier interactions. Here, AGP maintained the integrity of intestinal barrier in colitis mice. RNA-Sequencing results indicated that pathways related to anoikis (apoptosis induced by loss of cell-matrix interaction), mitochondrial function and oxidative stress were significantly altered in the process of AGP-intestinal barrier interaction. Further experiments confirmed that AGP activated Nrf2, decreased ROS levels, mitigated mitochondrial dysfunction and anoikis of colonic epithelial cells in mice. Intriguingly, AGP reversed oxidative stress and mitochondrial dysfunction induced by knockdown or inhibitor (ML385) of Nrf2 in IEC-6 cells, which indicated the essential role of Nrf2-mitochondrial axis in the intestinal protective function of AGP. In addition, AGP alleviated anoikis caused by impaired mitochondrial function. Hence, this current work indicated that AGP might maintain intestinal barrier integrity by mitigating anoikis mediated by Nrf2-mitochondria axis. These findings provide new evidence into the effect of polysaccharides maintaining intestinal barrier integrity.