The influence of sanguinarine(SA) on contraction of smooth muscle cells isolated from rat intestines using collagenase digestion was measured by inverted phase–contrast microscope. The result showed that rat intestinal smooth muscle cells isolated by enzyme digestion showed high purity, high density and “peak–valley” arrangement which is unique to smooth muscle cells. SA and atropine showed significant inhibition on the contractions of the isolated cells, the inhibition rates were 32.22% and 34.99% respectively. The inhibition rate of SA together with atropine on the contraction of the isolated cells was 68.8% which was significantly different from those when SA and atropine were used separately. SA showed significant inhibition on the contractions of intestinal smooth muscle cells induced by ACh, HA or KCl. The results suggested that the inhibition effects of SA on contraction of rat intestinal smooth muscle cells were achieved mainly through M receptor, H1 receptor and calcium ions channels on the cell membrane.