Introduction: Hypercortisolism, as seen in the majority of patients with major depression, may be associated with the generation of platelets that show signs of increased activation. Methods: Within a study using a placebo-controlled double-blind cross-over design, 9 healthy subjects ingested hydrocortisone (daily dose = 40 mg) or placebo on 7 consecutive days. At the end of each study segment, we analyzed platelets for the surface activation markers P-selectin, glycoprotein 53, and PAC-1 and measured platelet-leucocyte aggregates in serum. Results: Hydrocortisone ingestion was not associated with changes in the platelet activation markers P-selectin and PAC-1 or the number of circulating platelet-leucocyte aggregates but with a trend (p = 0.056) toward higher expression of glycoprotein 53 on the platelet surface. Conclusions: Induction of hypercortisolism in healthy volunteers was not associated with a major increase in platelet activation markers.