Abstract: 【Objective】To explore the mechanism of action of mogroside V（MV）in enhancing the developmental ability of cattle 8-cell embryos，which could provide theoretical basis for improving the developmental ability of cattle embryos and improving the quality of in vitro cultured embryos using plant extracts.【Method】Cattle 4/8/16-cell embryos were collected，and transcriptome sequencing was performed using the Smart-seq2 method to explore the transcriptional expression profile of mitochondrial-related pathways in cattle embryos，and mitochondrial staining verification，uniform manifold approximation and projection（UMAP）clustering and gene set enrichment analysis（GSEA）were conducted. Different concentrations（25，50，100 μmol/L）of MV were added during the in vitro culture of cattle 8-cell embryos， and the optimal MV concentration was screened by counting the 8-cell rate and blastocyst rate. Two treatment groups were set up，normal cattle embryos（CON）and the optimal MV concentration treatment was as experimental group（MV）， and immunofluorescence staining was used to detect mitochondrial function，ROS level，cell apoptosis level and DNA damage level in cattle 8-cell embryos.【Result】GSEA results showed that there were differences in the transcriptional expression profiles of mitochondrial-related pathways in cattle embryos at different developmental stages. Compared with the 8-cell embryos，the mitochondrial proteins and their complex pathways in the 4-cell and 16-cell embryos were significantly up-regulated（P<0.05，the same below）. Mitochondrial staining results showed that the mitochondrial fluorescence intensity of cattle 8-cell embryos was significantly lower than that at the 4-cell stage（P<0.01，the same below）and significantly lower than that at the 16-cell stage. Adding 50 μmol/L MV had the most significant effect on improving the developmental ability of cattle embryos. Compared with the CON group，the mitochondrial fluorescence intensity of cattle 8-cell embryos in the MV group was significantly increased，the ROS level was significantly decreased，the fluorescence intensity of Caspase-3 was significantly decreased，and the fluorescence intensity of γ-H2AX was significantly decreased. 【Conclusion】Compared with cattle 4-cell and 16-cell embryos，mitochondrial function in 8-cell embryos is reduced. During embryonic development，the addition of MV can enhance mitochondrial function and reduce oxidative stress，apoptosis and DNA damage levels in 8-cell embryos，thereby improving the embryonic development ability of cattle and alleviating the developmental block of 8-cell embryos.