There are 27 million cases of Salmonella Typhimurium (STM) reported worldwide annually, which have resulted in 217,000 deaths to date. Thus, there is an urgent requirement to develop novel antibacterial agents to target the multidrug-resistant strains of STM. We evaluated the inhibitory effect of the chloroform extracts of Atractylodes chinensis (Ac-CE) on the virulence of STM in vitro and develop it as a potential antibacterial agent. First, we determined the in vitro effects of Ac-CE on STM biofilm formation, and swimming, swarming, and adhesion to mucin. Further, we evaluated the effect of Ac-CE on the adhesion and invasion of STM at the gene level. Lastly, we evaluated the inhibitory effect of Ac-CE on STM infectivity at the cellular level. Ac-CE could attenuate both the adhesion and invasion abilities of STM in vitro. At the gene level, it could inhibit the expression of flagella, pilus, biofilm, SPI-1, and SPI-2 genes, which are related to the adhesion and invasion ability of STM in cells. Ac-CE significantly downregulated the expression of inflammatory cytokines and the TLR4/MyD88/NF-κB pathway in an STM infection cell model. It also significantly recovered the expression of intestinal barrier–related genes and proteins in intestinal cells that are damaged during STM infection. Ac-CE is effective as an antivirulence agent in alleviating STM infection. Although the main components of Ac-CE were analyzed.We have not demonstrated the antivirulence effect of the active ingredients in Ac-CE. And the antivirulence effect of Ac-CE and its active ingredients warrant further in vivo studies.