Black rice (Oryza sativa L.) is often associated with blood lipid control. This study systematically assessed the inhibition of cholesterol absorption in vitro by black rice and explored cholesterol-lowering compounds present in this rice. Our results indicated that black rice extracts (BRE), which were aqueous, ethanol extracts and a fraction of macroporous resin caused the reduction of cholesterol absorption by inhibiting pancreatic lipase, decreasing the micellar cholesterol solubility and suppressing cholesterol uptake in Caco-2 cells. The inhibitory activity was positively associated with anthocyanin (cyanidin-3-glucoside (Cy-3-G) and peonidin-3-glucoside (Pn-3-G)) contents of the extracts. Therefore, the cholesterol absorption inhibiting properties of anthocyanins were further explored. The IC50 values of Cy-3-G and Pn-3-G against pancreatic lipase were 42.53 ± 4.45 and 18.13 ± 4.22 μg mL(-1), respectively. Kinetic analysis suggested that the enzymatic inhibitory mode of Cy-3-G and Pn-3-G belonged to the competitive type. In mixed micelles, Cy-3-G and Pn-3-G dose dependently reduced the solubility of cholesterol. Meanwhile, a potential mechanism of cholesterol reduction by anthocyanins was investigated. Results showed that anthocyanins led to precipitation of cholesterol from micellar solution, which may induce the reduction of cholesterol. In Caco-2 cells, Cy-3-G and Pn-3-G (40 μg mL(-1)) exhibited a significant reduction in cholesterol uptake, and the degree of this reduction was almost the same as that observed in the group treated with Ezetimibe at the same concentration. These findings provide important evidence that anthocyanins may partly contribute to the inhibitory effects of black rice on cholesterol absorption, and thus may be applied for the prevention and treatment of hypercholesterolaemia.