OBJECTIVE: To discuss the factors that influence serum digoxin concentration (SDC), and to provide reference for the clinical safe, effective and reasonable use of digoxin. METHODS: A retrospective analysis was conducted to analyze medical information of 334 patients who received Digoxin tablets (0.125 mg, once a day, for consecutive 7 days or above) and determination of steady-state blood concentration in the First Affiliated Hospital of Kunming Medical University during Jan. 2014-Dec. 2016. One-way ANOVA was used to analyze the effects of patient’s age, blood potassium level and creatinine clearance rate on SDC. Chi-square test was used to analyze the effects of gender, cardiac function grading and drug combination on SDC. Binary Logistic regression analysis was conducted for the drug combination with significant effects. RESULTS: One-way ANOVA showed that there was statistical significance in SDC among patients with different creatinine clearance rates (<30 mL/min, 30~<70 mL/min, ≥70 mL/min)(P<0.05). By Chi-square test, patients with creatinine clearance rate≥70 mL/min (normal renal function) combined with nonsteroidal anti-inflammatory drugs (NSAIDs), patients with creatinine clearance rate<70 mL/min (renal insufficiency) combined with β blockers, proton pump inhibitors, torasemide, spironolactone, budesonide and theophylline had significance effects on SDC (P<0.05). Results of Binary Logistic regression analysis showed that combined with spironolactone was positively correlated with SDC in renal insufficiency patients, and combined with theophylline was negatively correlated with SDC. CONCLUSIONS: When the renal function is basically normal, except for can not be combined with NSAIDs, other pathophysiological factors and drug combination hardly affect the SDC. For patients with renal insufficiency, combined use of digoxin and spironolactone can increase the probability of excessive high SDC; combined use of digoxin and theophylline can reduce the probability of excessive high SDC.