In traditional Chinese herb medicine, Alismatis rhizoma has been used in treating edema, inflammation and increasing urine flow. Mechanism of Alismatis rhizoma¡¦s effect on these functions has not been elucidated. Since diuresis has been considered closely related to the reabsorption of sodium ion via the epithelium of tubule and collecting duct in kidney, we suspected that Alismatis rhizoma may influence transportation of salt and water. The measurement of short circuit current ( Isc ) has been used widely to estimate the ion transportation between mucosal and serosal side of epithelium. In the present experiment, we used the voltage ¡V clamp technique to demonstrate the effect of Alismatis rhizoma¡¦s extracts ( ARE ) on Isc and sodium ion conductance in frog skin. Our result showed that in control stage, the potential difference ( PD ) of frog skin is 64.81 ¡Ó 2.44 mV, the Isc is 59.82 ¡Ó 3.58 £gA / cm2 and the conductance is 1.09 ¡Ó 0.18 £gA / cm2 / mV. After ARE was applied to mucosal side of the frog skin, its Isc decrease from 62.63 ¡Ó 5.31 £gA / cm2 to 47.92 ¡Ó 5.41 £gA / cm2, which could further go down to 3.36 ¡Ó 1.06 £gA/cm2 by adding amiloride. Treating serosal side of frog skin with ARE decreased approximately 10% of its Isc. No apparent changes in conductance were observed by adding ARE to mucosal ( 0.98 ¡Ó 0.24 £gA / cm2 / mV ) or serosal side ( 0.96 ¡Ó 0.23 £gA / cm2 / mV ). Adding L-arginine ( the precursor of nitric oxide ) to the serosal side of the skin epithelium elevated the Isc for 17.10 ¡Ó 9.30 £gA/cm2. This effect can be inhibited by applying ARE or NG-L-nitro-arginine methyl ester ( L-NAME, NO synthase inhibitor ) before application of L-arginine. In summary, Alismatis rhizoma could affect Isc on both mucosal and serosal sides of the frog skin. Its effect on lowering Isc was more obvious when applied to the mucosal side than to the serosal side. The ARE may exert its effect on mucosal side by affecting amiloride¡Vsensitive sodium channel and on serosal side by affecting the NO signal transduction pathway.

Rhei Rhizoma, also named as rhubarb or Da Huang, has been used widely in oriental traditional medicine in treating constipation and edema. However, though much affection has been paid to the make of components on pharmaceutical mechanisms, few studies have been conducted to reveal chemical and physical mechanism of these effects. Studies have shown that diarrhea causes imbalance of chloride and sodium ion movements via epithelium, we wondered if similar mechanism may apply to Rhei Rhizoma, a herbal drug which has been used to treat constipation in oriental medicine for thousands of years. The measurement of short-circuit current (Isc) has been used widely to estimate the ion transportation between mucosal and serosal side of epithelium. In this study, we used Ussing chamber technique to examine the alternation in membrane potential and short-circuit currents. The result shows, at default, the Isc of frog skin we used was at 59.23¡Ó5.58£gA/cm², and the conductance was at 1.11¡Ó0.50£gA/cm²¡EmV. The lnjection of 1ml RRE to mucosal side of the frog skin leaded to a 90% elevation of the Isc. Followed by the application of Amiloride (sodium channel inhibitor) and Chlorothiazide (chloride channel inhibitor) to mucosal side of the epithelial skin, the observed Isc were then reduced 136% and 33% respectively. If RRE were applied after the adding of Amiloride or Chlorothiazide to the frog skin, then the Isc of the skin elevated only 24% and 70% respectively. These results show that Rhei Rhizoma Extract (RRE) significantly increases Isc upon application to the mucosal side of the skin epithelium. Amiloride and Chlorothiazide will both inhibit the Isc induced by RRE, indicating activation of chloride channel and Amiloride-sensitive sodium channel of the epithelial tissue by RRE. After the regular Ringer solution used in the preparation was replaced by Na-free and Cl-free Ringer solution, the inhibition of Isc by RRE application could still be observed although the inhibition was trivia. These results indicate that RRE acts dominantly on mucosa side of the epithelium and can be used to enhance sodium transport and to stimulate the secretion of Cl- in the epithelium.