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Cardiac fibrosis is more likely to promote the development of heart failure in the elderly. However, effective prevention and improvement of cardiac fibrosis are lacking. Here, we investigated the ameliorative effect of lactoferrin (LF) on aging cardiac fibrosis and its possible mechanism. We treated aged mice with LF and measured its effects on cardiac function, cardiac aging, cardiac fibrosis, and inflammatory response. LF effects on proliferation, transformation, collagen synthesis, and related signalling pathways of fibroblasts were observed in vitro, indicating that LF could delay heart aging, improve heart function, reduce fibrosis, and inhibit inflammation. LF inhibited proliferation, transformation, collagen synthesis, and inflammation associated with nuclear factor kappa-B (NF-κB) via 5′-AMP-activated protein kinase (AMPK) activation in myocardial fibroblasts, suggesting that LF ameliorates cardiac fibrosis and protects the heart in aged mice via AMPK activation. Thus, LF is a functional food for preventing and improving heart disease associated with cardiac fibrosis.