The present study investigated the underlying cellular mechanism in the effect of ligustrazine (tetramethylpyrazine, TMP) on the anion secretion of colonic mucosa in rats using a short-circuit current (I(sc)) technique in conjunction with "tool drugs." (i) After a pretreatment of the tissues by bathing the bilateral surface with Cl(-)-free Krebs-Henseleit (K-H) solution for over an hour, a basolateral application of 1 mmol/l TMP produced an increase in I(sc), and the total charges transported for 30 min were about 8.7 +/- 1.4 mC/cm(2); an apical pretreatment of DPC and a basolateral addition of acetazolamide decreased the TMP-induced I(sc) by about 60% (P < 0.01) and 45% (P < 0.05), respectively; a basolateral application of 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), the inhibitor of Na(+)-HCO(3)(-) cotransporter (NBC), did not alter the TMP-induced I(sc). (ii) After the bilateral surface of mucosa was bathed with HCO(3)(-)-free K-H solution for over an hour, a basolateral application of 1 mmol/l TMP produced an increase in I(sc), and the total charges transported in 30 min were about 8.3 +/- 1.9 mC/cm(2); an apical pretreatment of DPC (1 mmol/l), the inhibitor of Cl(-) channels, decreased the TMP-induced Isc by about 84% (P < 0.01). The basolateral presence of bumetanide (0.1 mmol/l), the inhibitor of Na(+)-K(+)-Cl(-) cotransporter (NKCC), significantly reduced the TMP-evoked I(sc) by about 86% (P < 0.01). In conclusion, (i) ligustrazine could promote colonic mucosa secretion Cl(-) via apical Cl(-) channels and basolateral NKCC; (ii) ligustrazine could promote colonic mucosa secretion HCO(3)(-) via apical Cl(-) channels and the basolateral diffusion of CO(2).