Abstract
Background and Aim: Local anesthetics which preferentially interact with voltage-gated sodium channels (VGSCs) were shown to have a clinical beneficial effect in ulcerative colitis and to regulate the secretion of inflammatory mediators from intestinal epithelial cells. However, expression of VGSCs in epithelial cells was not demonstrated. Herein we assessed whether intestinal epithelial cells express VGSCs. Methods: The expression of VGSCs in normal human colonic tissue and in the TNFα-treated or untreated intestinal epithelial cell line HT-29 was studied by immunofluorescence staining and FACS analysis, Western blot, immunohistochemistry, and RT-PCR using primers specific for several VGSC α subunits. The function of VGSCs was assessed by measuring changes in the membrane potential of the intestinal epithelial cells following incubation with lidocaine, veratridine, or both. Results: HT-29 cells were shown to express the VGSC α protein. mRNA analysis revealed the expression of nine of ten VGSC α isoforms. Immunohistochemical staining of normal colonic tissue confirmed the expression of VGSCs in colonic epithelial cells, smooth muscle cells, and nerves. Lidocaine induced membrane depolarization of HT-29 cells and its effect was blocked by veratridine. Conclusion: Malignant and normal intestinal epithelial cells express functional VGSCs. These molecules may play a role in the regulation of inflammation in gut physiology and pathology.
Journal Section:
Original Paper
References
1.
Uhlig HH, Powrie F: The role of mucosal T lymphocytes in regulating intestinal inflammation. Springer Semin Immunopathol 2005;27:167–180.
[PubMed]
2.
Inagaki A, Yamaguchi S, Ishikawa T: Amiloride-sensitive epithelial Na+ channel currents in surface cells of rat rectal colon. Am J Physiol Cell Physiol 2004;286:C380–C390.
[PubMed]
3.
Daig R, Rogler G, Aschenbrenner E, Vogl D, Falk W, Gross V, Scholmerich J, Andus T: Human intestinal epithelial cells secrete interleukin-1 receptor antagonist and interleukin-8 but not interleukin-1 or interleukin-6. Gut 2000;46:350–358.
[PubMed]
4.
Bjorck S, Jennische E, Dahlstrom A, Ahlman H: Influence of topical rectal application of drugs on dextran sulfate-induced colitis in rats. Dig Dis Sci 1997;42:824–832.
[PubMed]
5.
Lahav M, Levite M, Bassani L, Lang A, Fidder H, Tal R, Bar-Meir S, Mayer L, Chowers Y: Lidocaine inhibits secretion of IL-8 and IL-1beta and stimulates secretion of IL-1 receptor antagonist by epithelial cells. Clin Exp Immunol 2002;127:226–233.
[PubMed]
6.
Yu FH, Catterall WA: The VGL-chanome: a protein superfamily specialized for electrical signaling and ionic homeostasis. Sci STKE 2004;2004:re15.
7.
Li RA, Ennis IL, Tomaselli GF, Marban E: Structural basis of differences in isoform-specific gating and lidocaine block between cardiac and skeletal muscle sodium channels. Mol Pharmacol 2002;61:136–141.
[PubMed]
8.
Goldin AL: Diversity of mammalian voltage-gated sodium channels. Ann NY Acad Sci 1999;868:38–50.
[PubMed]
9.
Catterall WA: Structure and function of voltage-gated ion channels. Trends Neurosci 1993;16:500–506.
[PubMed]
10.
Ragsdale DS, McPhee JC, Scheuer T, Catterall WA: Common molecular determinants of local anesthetic, antiarrhythmic, and anticonvulsant block of voltage-gated Na+ channels. Proc Natl Acad Sci USA 1996;93:9270–9275.
[PubMed]
11.
Jo T, Nagata T, Iida H, Imuta H, Iwasawa K, Ma J, Hara K, Omata M, Nagai R, Takizawa H, Nagase T, Nakajima T: Voltage-gated sodium channel expressed in cultured human smooth muscle cells: involvement of SCN9A. FEBS Lett 2004;567:339–343.
[PubMed]
12.
Levite M, Cahalon L, Peretz A, Hershkoviz R, Sobko A, Ariel A, Desai R, Attali B, Lider O: Extracellular K(+) and opening of voltage-gated potassium channels activate T cell integrin function: physical and functional association between Kv1.3 channels and beta1 integrins. J Exp Med 2000;191:1167–1176.
[PubMed]
13.
Diebel LN, Liberati DM, Baylor AE 3rd, Brown WJ, Diglio CA: The pivotal role of tumor necrosis factor-alpha in signaling apoptosis in intestinal epithelial cells under shock conditions. J Trauma 2005;58:995–1001.
[PubMed]
14.
Wittmack EK, Rush AM, Hudmon A, Waxman SG, Dib-Hajj SD: Voltage-gated sodium channel Nav1.6 is modulated by p38 mitogen-activated protein kinase. J Neurosci 2005;25:6621–6630.
[PubMed]
15.
Catterall WA: From ionic currents to molecular mechanisms: the structure and function of voltage-gated sodium channels. Neuron 2000;26:13–25.
[PubMed]
16.
Catterall WA: Neurotoxins that act on voltage-sensitive sodium channels in excitable membranes. Annu Rev Pharmacol Toxicol 1980;20:15–43.
[PubMed]
17.
Rando TA, Wang GK, Strichartz GR: The interaction between the activator agents batrachotoxin and veratridine and the gating processes of neuronal sodium channels. Mol Pharmacol 1986;29:467–477.
[PubMed]
18.
Marban E, Yamagishi T, Tomaselli GF: Structure and function of voltage-gated sodium channels. J Physiol 1998;508:647–657.
[PubMed]
19.
Diss JK, Fraser SP, Djamgoz MB: Voltage-gated Na+ channels: multiplicity of expression, plasticity, functional implications and pathophysiological aspects. Eur Biophys J 2004;33:180–193.
[PubMed]
20.
Cummins TR, Howe JR, Waxman SG: Slow closed-state inactivation: a novel mechanism underlying ramp currents in cells expressing the hNE/PN1 sodium channel. J Neurosci 1998;18:9607–9619.
[PubMed]
21.
Cummins TR, Aglieco F, Dib-Hajj SD: Critical molecular determinants of voltage-gated sodium channel sensitivity to mu-conotoxins GIIIA/B. Mol Pharmacol 2002;61:1192–1201.
[PubMed]
22.
Akopian AN, Souslova V, England S, Okuse K, Ogata N, Ure J, Smith A, Kerr BJ, McMahon SB, Boyce S, Hill R, Stanfa LC, Dickenson AH, Wood JN: The tetrodotoxin-resistant sodium channel SNS has a specialized function in pain pathways. Nat Neurosci 1999;2:541–548.
[PubMed]
23.
Burgess DL, Kohrman DC, Galt J, Plummer NW, Jones JM, Spear B, Meisler MH: Mutation of a new sodium channel gene, Scn8a, in the mouse mutant ‘motor endplate disease’. Nat Genet 1995;10:461–465.
[PubMed]
24.
Planells-Cases R, Caprini M, Zhang J, Rockenstein EM, Rivera RR, Murre C, Masliah E, Montal M: Neuronal death and perinatal lethality in voltage-gated sodium channel alpha(II)- deficient mice. Biophys J 2000;78:2878–2891.
[PubMed]
25.
Black JA, Dib-Hajj S, McNabola K, Jeste S, Rizzo MA, Kocsis JD, Waxman SG: Spinal sensory neurons express multiple sodium channel alpha-subunit mRNAs. Brain Res Mol Brain Res 1996;43:117–131.
[PubMed]
26.
Boiko T, Van Wart A, Caldwell JH, Levinson SR, Trimmer JS, Matthews G: Functional specialization of the axon initial segment by isoform-specific sodium channel targeting. J Neurosci 2003;23:2306–2313.
[PubMed]
27.
Fraser SP, Salvador V, Manning EA, Mizal J, Altun S, Raza M, Berridge RJ, Djamgoz MB: Contribution of functional voltage-gated Na+ channel expression to cell behaviors involved in the metastatic cascade in rat prostate cancer. I. Lateral motility. J Cell Physiol 2003;195:479–487.
[PubMed]
28.
Breese EJ, Michie CA, Nicholls SW, Murch SH, Williams CB, Domizio P, Walker-Smith JA, MacDonald TT: Tumor necrosis factor alpha-producing cells in the intestinal mucosa of children with inflammatory bowel disease. Gastroenterology 1994;106:1455–1466.
[PubMed]
29.
Jung HC, Eckmann L, Yang SK, Panja A, Fierer J, Morzycka-Wroblewska E, Kagnoff MF: A distinct array of proinflammatory cytokines is expressed in human colon epithelial cells in response to bacterial invasion. J Clin Invest 1995;95:55–65.
[PubMed]
30.
Black JA, Liu S, Tanaka M, Cummins TR, Waxman SG: Changes in the expression of tetrodotoxin-sensitive sodium channels within dorsal root ganglia neurons in inflammatory pain. Pain 2004;108:237–247.
[PubMed]
31.
Gordienko DV, Tsukahara H: Tetrodotoxin-blockable depolarization-activated Na+ currents in a cultured endothelial cell line derived from rat interlobar arter and human umbilical vein. Pflügers Arch 1994;428:91–93.
32.
DeCoursey TE, Chandy KG, Gupta S, Cahalan MD: Voltage-dependent ion channels in T-lymphocytes. J Neuroimmunol 1985;10:71–95.
[PubMed]
33.
Bennett ES, Smith BA, Harper JM: Voltage-gated Na+ channels confer invasive properties on human prostate cancer cells. Pflügers Arch 2004;447:908–914.
34.
Holm AN, Rich A, Miller SM, Strege P, Ou Y, Gibbons S, Sarr MG, Szurszewski JH, Rae JL, Farrugia G: Sodium current in human jejunal circular smooth muscle cells. Gastroenterology 2002;122:178–187.
[PubMed]
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