Protective effects of chlorogenic acid against LPS-induced intestinal oxidative injury in mice via activation of the PI3K/Akt-Nrf2/HO-1 signaling axis

Introduction Oxidative stress is a key pathological cause of intestinal epithelial injury and related intestinal diseases. Chlorogenic acid (CGA), a natural polyphenol, has notable antioxidant activity. This study explored the multi-target protective effects of CGA on intestinal epithelial cells and mouse intestinal tissue against oxidative injury, as well as its underlying molecular mechanisms. Methods Network pharmacology was applied to screen core targets and enriched pathways of CGA against oxidative stress. H₂O₂-induced MODE-K cell oxidative damage model and LPS-induced mouse intestinal injury model were constructed. CCK-8, biochemical detection, qPCR, molecular docking, Western blotting and histopathological assay were performed to verify the protective effects and related signaling pathways. Results Network pharmacology predicted PI3K/Akt and Nrf2/HO-1 pathways as crucial targets. In vitro , CGA elevated cell viability, reduced oxidative injury markers, upregulated antioxidant factors, and regulated apoptosis-related proteins. CGA blocked Keap1-Nrf2 binding, activated Nrf2/HO-1 and promoted Akt phosphorylation. In vivo , CGA alleviated LPS-induced intestinal pathological injury, while PI3K inhibitor LY294002 reversed its protective effects, confirming the critical role of PI3K/Akt-Nrf2/HO-1 axis. Conclusion CGA exerts multi-target antioxidant and intestinal protective effects by inhibiting Keap1-Nrf2 interaction and activating PI3K/Akt-Nrf2/HO-1 signaling axis, mitigating intestinal oxidative injury. Our findings provide a mechanistic basis for the application of CGA as a functional food ingredient or dietary intervention to alleviate intestinal oxidative stress in livestock.