Objective To investigate the effect of Yantiao Fang on apoptosis of intestinal epithelial cells in mice with acute gastrointestinal injury caused by sepsis by regulating the Rho / ROCK signaling pathway. Methods Seventy BALB / c mice were randomly divided into normal, sham, and model groups. Except for normal and sham groups, mice were subjected to cecal ligation and perforation to establish a mouse model of acute gastrointestinal injury caused by sepsis.The mouse models were randomly divided into model, Low, Medium, and High dose of Yantiao Fang, and ROCK inhibitor groups. Histopathological changes of the ileum were observed by HE staining. Serum levels of IL-1β, IL-6, TNF-α, and IL-10 were measured by ELISA. PCNA and Ki-67 expression was detected by immunohistochemistry. Cleaved caspase3 and Bax expression was detected by Western blot. ROCK and MLC mRNA expression in the ileum was measured by RTqPCR. Results Compared with normal and sham groups, Chiu’ s pathological score, proinflammatory factor ( IL-1β, IL6, and TNF-α) levels, cleaved caspase3 and Bax protein expression, and ROCK and MLC mRNA levels were increased in the model group (P<0. 05) . Moreover, anti-inflammatory mediator IL-10 and expression of PCNA and Ki-67 in the ileum were decreased in the model group (P<0. 05) . Compared with the model group, histopathological changes of the ileum in all Yantiao Fang groups were improved by various degrees with the increase in dose. Chiu’ s pathological score, IL-1β, IL6, and TNF-α serum levels, cleaved caspase3 and Bax protein expression, and ROCK and MLC mRNA levels were decreased in Yantian Fang groups ( P< 0. 05) . The IL-10 level and expression of PCNA and Ki-67 in the ileum were increased in Yantian Fang groups ( P< 0. 05) . Conclusions Yantiao Fang may inhibit apoptosis of intestinal epithelial cells in mice with acute gastrointestinal injury due to sepsis by regulating the Rho / ROCK signaling pathway, thereby alleviating intestinal tissue inflammation and ultimately preventing intestinal mucosal tissue injury.