To investigate the pharmacological mechanism of the traditional Chinese medicine, Pulsatilla decoction (PD), the levels of nitric oxide (NO), endothelin-1 (ET-1), tumor necrosis factor-alpha (TNF-alpha), and interleukin-1 alpha (IL-1 alpha) secreted by cultured rat intestinal microvascular endothelial cells (RIMECs) were determined after treatment with PD and its seven active ingredients, namely anemoside B(4), anemonin, berberine, jatrorrhizine, palmatine, aesculin, and esculetin. RIMECs were challenged with lipopolysaccharide (LPS) at 1 microg ml(-1) for 3 h and then treated with PD at 1, 5, and 10 mg ml(-1) and its seven ingredients at 1, 5, and 10 microg ml(-1) for 21 h, respectively. The results revealed that PD, anemonin, berberine, and esculetin inhibited the production of NO; PD, anemonin, and esculetin inhibited the secretion of ET-1; PD, anemoside B(4), berberine, jatrorrhizine, and aesculin downregulated TNF-alpha expression; PD, anemoside B(4), berberine, and palmatine decreased the content of IL-1 alpha. It showed that PD and its active ingredients could significantly inhibit the secretion of NO, ET-1, TNF-alpha, and IL-1 alpha in LPS-induced RIMECs and suggested they would reduce inflammatory response via these cytokines.