Clin Exp Otorhinolaryngol.  2008 Jun;1(2):75-79. 10.3342/ceo.2008.1.2.75.

Modulators of Ion Transport in Nasal Polyps: An in situ Measurement of Short-Circuit Current

• Lee JH
• Rhee CS
• Kim DW
• Lee CH

• KMID: 1486057
• DOI: http://doi.org/10.3342/ceo.2008.1.2.75

Abstract

OBJECTIVES
To examine possible modulators of the ion transport through the apical membrane of the nasal polyps. METHODS: The study was conducted using the freshly-excised nasal polyps from the patients with chronic sinusitis. A voltage-sensitive vibrating probe technique was introduced to monitor the short-circuit current across the apical membrane of the polyp at 37degrees C. RESULTS: In the presence of amiloride, Adenosine 5'-triphosphate induced 4,4'-Diisothiocyanatostilbene-2,2'-disulfonic acidsensitive chloride current. Uridine 5'-diphosphate was less potent than Uridine 5'-triphosphate, and adenosine increased chloride secretion, which was blocked by the antagonist, 8-(p-sulfophenyl) theophylline on adenosine receptor. Based on the pharmacologic profiles, multiple purinergic receptors, including P2Y(2), P2Y(6), and P1 receptors, were functionally expressed. However, P2X receptor agonists (alpha,beta-methyleneadenosine 5'-triphosphate and 2'- & 3'-O-[4-benzoyl-benzoyl] adenosine 5'-triphosphate), Cystic fibrosis conductance regulator (CFTR) activator (genistein), nitric oxide substrate (L-arginine), and nitric oxide donor (sodium nitroprusside) had no significant effect on the short circuit current. CONCLUSION: Among tested drugs, P2Y receptor agonists were major modulators of ion transport in nasal polyps in situ.

Keyword

Nasal polyps; CFTR; Purinergic receptors; Genistein; Nitric oxide; Vibrating Probe

MeSH Terms

Adenosine
Amiloride
Cystic Fibrosis
Genistein
Humans
Ion Transport
Membranes
Nasal Polyps
Nitric Oxide
Organothiophosphorus Compounds
Polyps
Receptors, Purinergic
Receptors, Purinergic P1
Sinusitis
Theophylline
Tissue Donors
Uridine
Adenosine
Amiloride
Genistein
Nitric Oxide
Organothiophosphorus Compounds
Receptors, Purinergic
Receptors, Purinergic P1
Theophylline
Uridine

Figure

• 

  Fig. 1 Effects of DIDS, DPC, and ATP in the presence of amiloride on the Isc. (A) A raw trace representative of 17 experiments was shown. The majority of the remained Isc in the presence of amiloride is accounted for by DPC-sensitive chloride conductance. (B) Effects of DIDS and DPC on the Isc after activation of chloride secretion by ATP in the presence of amiloride. A raw trace representative of 7 experiments was shown.DPC: N-phenylanthranilic acid; DIDS: 4,4'-Diisothiocyanatostilbene-2,2'-disulfonic acid disodium salt hydrate.

• 

  Fig. 2 Dose-response relationships of ATP (A) and UTP (B) in the presence of amiloride. (C) EC50 of ATP (dashed line) and UTP (continuous line) was 5.3 and 7.3 µM, respectively (n=5).

• 

  Fig. 3 Effects of ATP, UTP, UDP, and adenosine on the Isc. (A) A representative trace of 5 experiments. UDP acting on P2Y6 receptors was less potent than UTP on P2Y2 receptors. UDP was preincubated with hexokinase in the presence of glucose to eliminate small amount of UTP contamination. (B) Effect of adenosine in the presence of amiloride on the Isc. Adenosine (Ad) also activated chloride secretion, which was inhibited by a nonspecific blocker (8-SPT).Ad: Adenosine; 8-SPT: 8-(p-Sulfophenyl) theophylline.

• 

  Fig. 4 Effects of P2X agonists, genistein (A), and nitric oxide donors (B) in the presence of amiloride on the Isc. None of these agents had an effect on the nasal polyps in situ. (A) αβmeATP, BzATP (100 µM), and genistein (20 µM) were used. (B) L-arginine (L-arg, 1 mM), sodium nitroprusside (SNP, 1 mM), and ATP (100 µM) were used.Alpha-beta-meATP: Alpha-beta-methyleneadenosine 5'-triphosphate; BzATP: 2'- & 3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate.

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