Bacterial diarrhea remains a significant global challenge in livestock farming, leading to serious animal health issues. The effects of Tupistra chinensis Baker therapy on intestinal alternations in Escherichia coli (E. coli)-infected mice are evaluated in this study. Thirty ICR mice were randomly divided into three groups: CK, PK, and MK. Both the CK and MK groups received normal saline intragastrical for 14 days, while the PK group was administered TCB polysaccharide solution for the same period. On 14^th day, the CK group was given PBS intragastrical while, both the MK and PK groups received an E. coli solution (8×10⁹ CFU). Serum inflammatory factors and oxidative stress indexes were determined by kits. Total fecal DNA was extracted for amplification. While amplified products were processed for sequencing by MiSeq sequencer. The histopathology analysis revealed that the intestinal injury in the PK group was mitigated, with increased villi height and less crypt depth compared to the MK group. The serum levels of inflammatory cytokines IL-1β, IL-6, and IL-10 were significantly restored in the PK group compared to the MK group (P<0.0001). Moreover, the antioxidant capacity was enhanced significantly, as indicated by increased SOD and GSH-px level. Additionally, intestinal microbiota sequencing identified 25 distinct genera, including butyrate-producing bacteria such as Dysosmobacter (P<0.05), Angelakisella (P<0.05), Lachnoclostridium_B (P<0.01), and Eubacterium_J (P<0.01). Conversely, pathogenic genera such as Alloprevotella (P<0.05), Adlercreutzia (P<0.05), and Muribaculum (P<0.05) were reduced in the PK group. In conclusion, TCB has the potential to protect the gut from oxidative stress and inflammation caused by E. coli infection.