Involvement of intracellular and extracellular Ca2+ in tetramethylpyrazine-induced colonic anion secretion.
In the present study we investigated the role of Ca(2+) in tetramethylpyrazine (TMP)-induced anion secretion in the human colonic epithelial cell line, Caco-2, using the short-circuit current (I(SC)) technique in conjunction with intracellular Ca(2+) measurements. The results showed that TMP-induced I(SC) response was significantly reduced by 58.8% and 38.3% after inhibiting Ca(2+) ATPase of endoplasmic reticulum (ER) with thapsigargin and mobilizing ER stored Ca(2+) release with ATP, respectively. Conversely, thapsigargin- and ATP-evoked I(SC) responses were also significantly reduced by pretreatment with TMP by 43.2% and 38.5%, respectively. Conversely, removal of extracellular Ca(2+), apical but not basolateral, or the presence of the Ca(2+) chelator (EGTA) significantly increased TMP-induced I(SC) by 47.1% and 37.8%, respectively. Similar to TMP, thapsigargin-induced current increase was also enhanced by chelating extracellular Ca(2+) or in Ca(2+) free solution; however, removal of extracellular Ca(2+) did not significantly affect 3-isobutyl-1-methylxanthine (IBMX)- and forskolin-induced transepithelial current. Measurement of the intracellular concentration of free Ca(2+) ([Ca(2+)](i)) with fura-2/AM showed that TMP could induce an increase in [Ca(2+)](i) but pretreatment with TMP significantly reduced thapsigargin-evoked, but not ATP-induced, [Ca(2+)](i) increase. These results suggest that the effect of TMP on colonic anion secretion is partly mediated by TMP-increased [Ca(2+)](i) by acting on a target similar to thapsigargin. The observed inhibitory effect of extracellular Ca(2+) on Ca(2+)-dependent anion secretion represents a novel mechanism by which Ca(2+)-dependent regulation of epithelial electrolyte transport may be fine-tuned by extracellular Ca(2+) in the apical domain.
Zhu
Jin-Xia
JX
Department of Physiology, Basic Medical College, Capital University of Medical Sciences, Beijing, PR China.
Zhang
Gui-Hong
GH
Yang
Ning
N
Connie Wong
Hau-Yan
HY
Chung
Yiu-Wa
YW
Chan
Hsiao-Chang
HC