Embryonic stem cell telomere-mitochondrial peptides (EST-Mips) are a group of peptide molecules derived from bovine embryonic stem cells, closely associated with telomere maintenance and mitochondrial function. In this study, based on 736 EST-Mips-regulated protein targets provided by users, we employed network pharmacology and bioinformatics approaches to systematically elucidate their molecular mechanisms in delaying the aging process. By screening target-aging associations using platforms such as PubChem, GeneCards, Human Aging Genomic Resources (HAGR/GenAge), Open Genes, and AgeAnno, we identified a total of 179 shared aging-related targets (accounting for 24.3% of all targets). Key targets include hub proteins with node connectivity ≥ 50, such as TP53, AKT1, SRC, EGFR, TNF, IL6, MAPK1, and MAPK3. GO functional enrichment analysis revealed that EST-Mips primarily participate in cell cycle regulation, apoptosis signaling, oxidative stress response, and inflammatory responses. KEGG pathway analysis demonstrated that EST-Mips exert their anti-aging effects by modulating aging-related signaling pathways, including PI3K-Akt, FoxO, p53, MAPK, JAK-STAT, and NF-κB. Comprehensive analysis indicates that EST-Mips exert their anti-aging effects through a multidimensional network involving “telomere maintenance—mitochondrial homeostasis—metabolic remodeling—immune modulation” synergistically.