The purpose of the present investigation is to study how berberine (Ber) affect the intracellular free calcium concentration ( Ca(2+) (i)) of the smooth muscle cells of guinea pig colon by means of biwavelength spectrophotometry with Fura 2 AM in a cell suspension specimen. In the resting state, Ca(2+) (i) in HEPES Ringer solution (CaCl2 1.5 mmol/L) was 108 9.4 nmol/L (n=7), which was not significantly affected by Ber. On the other hand, Ber inhibited the increased Ca(2+) (i) induced by 60 mmol/L KCl in a dose dependent manner, with a value of IC(50) being 34.09 micromol/L. 30, 100 micromol/L Ber also inhibited 10 micromol/L ACh evoked Ca(2+) (i) elevation in a dose dependent manner either in the presence or absence of extracellular Ca(2+). In addition, Ber inhibited cyclopiazonic acid (CPA) stimulated Ca(2+) (i) elevation dose dependently. This effect was more potent in HEPES Ringer solution (IC(50)=37.79 micromol/L) than in Ca(2+) free medium (IC(50)=49.70 micromol/L). The above results suggest that Ber exerts an inhibitory effect on the extracellular Ca(2+) influx and the Ca(2+) release from intracellular stores.