Korean J Physiol Pharmacol.  2014 Aug;18(4):341-346. 10.4196/kjpp.2014.18.4.341.

Effects of Lubiprostone on Pacemaker Activity of Interstitial Cells of Cajal from the Mouse Colon

• Jiao HY
• Kim DH
• Ki JS
• Ryu KH
• Choi S
• Jun JY

• KMID: 2285529
• DOI: http://doi.org/10.4196/kjpp.2014.18.4.341

Abstract

Lubiprostone is a chloride (Cl-) channel activator derived from prostaglandin E1 and used for managing constipation. In addition, lubiprostone affects the activity of gastrointestinal smooth muscles. Interstitial cells of Cajal (ICCs) are pacemaker cells that generate slow-wave activity in smooth muscles. We studied the effects of lubiprostone on the pacemaker potentials of colonic ICCs. We used the whole-cell patch-clamp technique to determine the pacemaker activity in cultured colonic ICCs obtained from mice. Lubiprostone hyperpolarized the membrane and inhibited the generation of pacemaker potentials. Prostanoid EP1, EP2, EP3, and EP4 antagonists (SC-19220, PF-04418948, 6-methoxypyridine-2-boronc acid N-phenyldiethanolamine ester, and GW627368, respectively) did not block the response to lubiprostone. L-NG-nitroarginine methyl ester (L-NAME, an inhibitor of nitric oxide synthase) and 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ, an inhibitor of guanylate cyclase) did not block the response to lubiprostone. In addition, tetraethylammonium (TEA, a voltage-dependent potassium [K+] channel blocker) and apamin (a calcium [Ca2+]-dependent K+ channel blocker) did not block the response to lubiprostone. However, glibenclamide (an ATP-sensitive K+ channel blocker) blocked the response to lubiprostone. Similar to lubiprostone, pinacidil (an opener of ATP-sensitive K+ channel) hyperpolarized the membrane and inhibited the generation of pacemaker potentials, and these effects were inhibited by glibenclamide. These results suggest that lubiprostone can modulate the pacemaker potentials of colonic ICCs via activation of ATP-sensitive K+ channel through a prostanoid EP receptor-independent mechanism.

Keyword

Colon; Interstitial cells of Cajal; Lubiprostone; Pacemaker potentials

MeSH Terms

Alprostadil
Animals
Apamin
Calcium
Colon*
Constipation
Glyburide
Interstitial Cells of Cajal*
Membranes
Mice*
Muscle, Smooth
Nitric Oxide
Patch-Clamp Techniques
Pinacidil
Potassium
Tetraethylammonium
Lubiprostone
Alprostadil
Apamin
Calcium
Glyburide
Nitric Oxide
Pinacidil
Potassium
Tetraethylammonium

Figure

• 

  Fig. 1 Cultured interstitial cells of Cajal (ICCs) from the mouse colon. The tunica muscularis of the colon was digested with collagenase, and the dispersed cells were cultured for 2 days. Confocal microscopic image of Kit-immunopositive ICCs network in culture.

• 

  Fig. 2 Effects of lubiprostone on pacemaker potentials in cultured interstitial cells of Cajal (ICCs) from the mouse colon. (A~C) Pacemaker potentials from ICCs exposed to lubiprostone (10 nM to 1 µM) in current clamping. Lubiprostone caused a concentration-dependent hyperpolarization of the membrane and inhibited the generation of pacemaker potentials. The responses to lubiprostone are summarized in D and E. Bars represent mean values±standard error (SE). *(p<0.05) Significantly different from the untreated control. The dotted lines indicate the resting membrane potentials.

• 

  Fig. 3 Effects of prostanoid EP receptor antagonists on lubiprostone-induced responses of pacemaker potentials in cultured interstitial cells of Cajal (ICCs) of the mouse colon. (A~D) Pacemaker potentials from ICCs exposed to lubiprostone (100 nM) in the presence of prostanoid EP receptor antagonists. SC-19220 (an EP1 receptor antagonist; 5 µM), PF-04418948 (an EP2 receptor antagonist; 10 µM), 6-methoxypyridine-2-boronc acid N-phenyldiethanolamine ester (an EP3 receptor antagonist; 10 µM), and GW627368 (an EP4 receptor antagonist; 10 µM) did not block the lubiprostone-induced responses on pacemaker potentials. The responses to lubiprostone in the presence of EP receptor antagonists are summarized in (E) and (F). Bars represent mean values±standard error (SE). The dotted lines indicate the resting membrane potentials (SC, SC-19220; PE, PF-04418948; EP3A, 6-methoxypyridine-2-boronc acid N-phenyldiethanolamine ester; GW, GW627368).

• 

  Fig. 4 Effects of L-NG-nitroarginine methyl ester (L-NAME) and H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ) on lubiprostone-induced responses on pacemaker potentials in cultured interstitial cells of Cajal (ICCs) of the mouse colon. (A, B) Pacemaker potentials from ICCs exposed to lubiprostone (100 nM) in the presence of L-NAME and ODQ. L-NAME (100 µM) and ODQ (10 µM) depolarized the membrane and increased the pacemaker potential frequency. However, lubiprostone continued to inhibit the pacemaker potentials in the presence of L-NAME and ODQ. The responses to lubiprostone in the presence of L-NAME and ODQ are summarized in (C) and (D). Bars represent mean values±standard error (SE). The dotted lines indicate the resting membrane potentials.

• 

  Fig. 5 Effects of tetraethylammonium (TEA) and apamin on lubiprostone-induced responses on pacemaker potentials in cultured interstitial cells of Cajal (ICCs) of the mouse colon. (A) TEA (5 mM) and (B) apamin (100 nM) did not block the effects of lubiprostone (100 nM) on pacemaker potentials. The responses to lubiprostone in the presence of TEA and apamin are summarized in (C) and (D). Bars represent mean values±standard error (SE). The dotted lines indicate the resting membrane potentials.

• 

  Fig. 6 Effects of glibenclamide on lubiprostone-induced responses and pinacidil effects on pacemaker potentials in cultured interstitial cells of Cajal (ICCs) of the mouse colon. (A) Glibenclamide (10 µM) inhibited the lubiprostone-induced responses on pacemaker potentials. (B) Pinacidil (100 nM) hyperpolarized the membrane and inhibited the pacemaker potentials, which were blocked by glibenclamide. The responses to glibenclamide in the lubiprostone- or pinacidil-induced effects are summarized in (C) and (D). Bars represent mean values±standard error (SE). The dotted lines indicate the resting membrane potentials (pina, pinacidil).

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