Saponins are the major chemicals from Rosa laevigata Michx fruit. However, there have no papers to report the activities of the natural product against liver damage. In the present work, total saponins from R. laevigata Michx fruit (RLTS) with purity>70% was produced and then the protective effects of it against CCl4-induced acute liver injury in mice was tested. The results showed that RLTS decreased serum ALT and AST activities compared with model group, as well as the relative liver weight and histological findings. In addition, RLTS remarkably increased the levels of SOD, CAT, GSH-Px, GSH and decreased MDA, iNOS and NO levels in liver. Transmission electron microscopy and TUNEL assays showed that RLTS repaired fragmented DNA and mitochondrial change caused by CCl4. Further investigation demonstrated that RLTS pretreatment down-regulated the protein expression of CYP2E1, ATF6, GRP78, EIF2, COX-2, NF-κB, p53, Caspase-3, Caspase-9, Cytokeratin 18 and the levels of MAPKs phosphorylation, up-regulated Bcl-2 expression, and markedly decreased the gene expression of TNF-α, IL-6, Fas/FasL and Bax. This is the first time to reveal the hepatoprotective effect of total saponins from R. laevigata Michx fruit, which should be developed as a new drug for treatment of live injury in future.