Objective The main aim of the study was to verify that digoxin was able to cause peptic ulcer. Methods Digoxin injection was prepared by dissolving digoxin powder in normal saline for injection containing 25% alcohol. Kunming mice were subcutaneously injected with normal saline for injection containing 25% alcohol， or with digoxin of 10， 20 and 40 mg/kg， respectively. In an oral administration experiment， mice were administrated with normal saline， or with oral digoxin solution of 10 and 20 mg/kg via free drink， respectively. Mice were treated for 5 days， and their body weight was recorded. In the end of the experiment， animals were killed， and acidity of the stomach and duodenum juice was detected. The general morphology of gastric antrum and duodenum was observed， and then， the tissues were fixed in 4% methanal， embedded in paraffin， and stained with hematoxylin-eosin. Results Mice injected with digoxin grew more slowly than those in control group， pH of stomach juice in mice treated with digoxin of 10 and 20 mg/kg significantly decreased， and mucosa of gastric antrum and duodenum congested. In the oral experiment， when mice administrated with digoxin solution， their body weight decreased， pH of stomach juice in the mice treated with digoxin of 10 mg/kg significantly decreased， mucosa of gastric antrum and duodenum also congested， and especially， mucosa of duodenum bled obviously. Conclusion Digoxin was able to induce peptic ulcer， and the effect of the drug administrated through oral was more powerful than that through injection.