The aim of present study was to investigate the effect of baicalin on hypoxia/reoxygenation (H/R) injury in cardiomyocytes and the mechanisms involved, particularly in relation to cytokines. The cardiomyocytes for the H/R groups were placed into a hypoxic chamber for 12 h and then underwent reoxygenation for 1 h. The cells in the TNF- groups were administered 100 ng/mL rrTNF- and incubated for 13 h under normal conditions. The cells in the baicalin pretreatment groups were administered 10 μM baicalin 30 min prior to exposure to H/R or TNF-. It was observed that pretreatment with baicalin (10 μM) significantly reduced the cell damage and death induced by H/R or TNF-. In the culture medium of the H/R cells, the SOD activity increased, while TNF- was decreased by baicalin. The levels of IL-6 in culture medium for H/R or TNF- treated cells were suppressed by baicalin pretreatment. In contrast, the levels of IL-10 in culture medium for H/R or TNF- treated cells were significantly elevated by baicalin. Moreover, baicalin inhibited the nuclear translocation of NF-κB induced by H/R or TNF-α. In conclusion, baicalin may protect cardiomyocytes from H/R injury through an anti-inflammatory mechanism.