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Induction of Mineralocorticoid Receptor by Sodium Butyrate in Small Intestinal (IEC6) and Colonic (T84) Epithelial Cell Lines

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Abstract

Mineralocorticoid action is essential forcolonic sodium and water absorption and is mediated viamineralocorticoid receptors in the upper half of coloniccrypts. On the other hand, it has been established that sodium butyrate inducesdifferentiation-like phenomenon in vitro. The aim ofthis study is to investigate whether this bacterialproduct participates in the regulation of gene andprotein expression of mineralocorticoid receptor in vitro. IEC6and T84 cells were stimulated by sodium butyrate andRNAs extracted. Gene expression of mineralocorticoidreceptor was evaluated by northern blotting orsemiquantitative RT-PCR. Protein expression was determined inT84 cells using immunohistochemistry. To investigatewhether MR induction was associated with cellulardifferentiation, we also measured alkaline phosphatase in situ. The mineralocorticoid receptor genewas induced by sodium butyrate in both IEC6 and T84cells. Immunoreactivity increased in butyrate-treatedT84 cells, but receptorcontaining cells were notuniformly distributed and often formed clusters.Induction of alkaline phosphate activity was alsodemonstrated in both IEC6 and T84 cells. Double stainingby immunoreactivity and alkaline phosphatase activityclearly demonstrated the colocalization of both afterbutyrate treatment. In conclusion, sodium butyrateup-regulates gene and protein expression of thefunctionally important mineralocorticoid receptor inepithelial cells, after induction by differentiation-likecondition in vitro.

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REFERENCES

  1. Mirshai M, Pagano M, Razaghi A, Lazar G, Agarwal MK: Immunophotochemical analysis of mineralocorticoid receptor by polyclonal antibodies against the native receptor from rat kidney. Biochem Med Metab Biol 47:133–144, 1992

    Google Scholar 

  2. Sasano H, Fukushima K, Sasaki I, Matsuno S, Nagura H, Krozowski ZS: Immunolocalization of mine ralocorticoid receptor in human kidney, pancreas, salivary, mammary and sweat glands. J Endocrinol 132:305–310, 1992

    Google Scholar 

  3. Rundle SE, Smith AI, Stockman D, Funder JW: Immunocytochemical demonstration of mineralocorticoid receptors in rat and human kidney. J Steroid Biochem 33:1235–1242, 1989

    Google Scholar 

  4. Krozowski ZS, Rundle SE, Wallace C, Castell MJ, Shen JH, Dowling J, et al: Immunolocalization of renal mineralocorticoid receptors with an antiserum against peptide deduced from the complementary deoxyribonucleic acid sequence. Endocrinology 125:192–198, 1989

    Google Scholar 

  5. Hirasawa G, Sasano H, Takahashi K, Fukushima K, Suzuki T, Hiwatashi N, et al: Colocalization of 11β-hydroxysteroid dehydrogenase type II and mineralocorticoid receptor in human epithelia. J Clin Endocrinol Metab 82:3859–3863, 1997

    Google Scholar 

  6. Lever JE: Inducers of mammalian cell differentiation stimulate dome formation in a differentiated kidney epithelial cell line (MDCK). Proc Natl Acad Sci USA 76:1323–1327, 1979

    Google Scholar 

  7. Wice BM, Trugnam G, Pinto M, Rousset M, Dussaulx GC, Lacroix B, et al: The intracellular accumulation of UDP-Nacetylhe xosamines is concomitant with the inability of human colon cancer cells to differentiate. J Biol Chem 260:139–146, 1985

    Google Scholar 

  8. Augeron C, Laboisse CL: Emerge nce of permanently differentiated cell clones in a human colonic cancer cell line in culture after treatment with sodium butyrate. Cancer Res 43:3961–3969, 1984

    Google Scholar 

  9. Tsao D, Morita A, A BJ, Luu P, Kim YS: Differential effects of sodium butyrate, dime thyl sulfoxide, and retinoic acid on membrane-associated antigen, enzymes, and glycoproteins of human rectal adenocarcinoma cells. Cancer Res 42:1052–1058, 1982

    Google Scholar 

  10. Tsao D, Shi Z, Wong A, Kim YS: Effect of sodium butyrate on carcinoembryonic antigen production by human colonic adenocarcinoma cells in culture. Cancer Res 43:1217–1222, 1983

    Google Scholar 

  11. Imondi AR, Balis ME, Lipkin M: Changes in enzyme levels accompanying differentiation of intestinal epithelial cells. Exp Cell Res 58:323–330, 1969

    Google Scholar 

  12. Hass R, Busche R, Luciano L, Reale E, Engelhardt RV: Lack of butyrate is associated with induction of Bax and subsequent apoptosis in the proximal colon of guinea pig. Gastroenterology 112:875–881, 1997

    Google Scholar 

  13. Utsunomiya J, Iwama T, Imajo M, Matsuo S, Sawai S, Yae-gashi K, et al: Total colectomy, mucosal proctectomy, and ileoanal anastomosis. Dis Colon Rectum 23:459–466, 1980

    Google Scholar 

  14. Will PC, Lebowitz JL, Hopfer U: Induction of amiloridesensitive sodium transport in the rat colon by mineralocorticoids. Am J Physiol 238:F261-F268, 1980

    Google Scholar 

  15. Will PC, Cortright RN, Groseclose RG, Hopfer U: Amiloridesensitive salt and fluid absorption in small intestine of sodium depleted rats. Am J Physiol 248:G133-G141, 1985

    Google Scholar 

  16. Rossier BC, Palmer LG: Mechanism of aldosterone action on sodium and potassium transport. In: The Kidney: Physiology and Pathophysiology. DW Seldin, G Giebisch (eds). New York, Raven Press, 1992, pp 1372–1409

    Google Scholar 

  17. Quaroni A, Wands J, Trelstad RL, Isselbacher KJ: Epithelioid cell cultures from rat small intestine: Characterization by morphologic and immunologic criteria. J Cell Biol 80:248–265, 1979

    Google Scholar 

  18. Mowles J, Moran S, Doyle A: Mycoplasma control. Nature 340:352, 1989 (letter, comment)

    Google Scholar 

  19. Fukushima K, Sasaki I, Takahashi K, Naito H, Ogawa H, Sato S, et al: Lipopolysaccharide exhibits synergistic enhancement of butyrate-induced and retinoic acid-mediated alkaline phosphatase activity on small intestinal epithelial cell line, IEC6. Digestion 59:683–688, 1998

    Google Scholar 

  20. Sambrook J, Fritsch EF, Maniatis T (eds): Molecular Cloning, A Laboratory Manual, 2nd ed. Plainview, New York, Spring Harbor Laboratory Press, 1989

    Google Scholar 

  21. Roland BL, Krozowski ZS, Funder JW: Glucocorticoid receptor, mineralocorticoid receptors, 11 beta-hydroxysteroid dehydrogenase-1 and-2 expression in rat brain and kidney: In situ studies. Mol Cell Endocrinol 111:R1-R7, 1995

    Google Scholar 

  22. Tetsuka T, Daphna-Iken D, Srivastava SK, Baier LD, Du-Maine J, Morrison AR: Cross-talk between cyclooxyge nase and nitric oxide pathways; Prostaglandin E2 negatively modulates induction of nitric oxide synthase by interleukin 1. Proc Natl Acad Sci USA 91:12168–12172, 1994

    Google Scholar 

  23. Arriza JL, Weinberger C, Cerelli G, Glaser TM, Handelin BL, Housman DE, et al: Cloning of human mine ralocorticoid receptor complementary DNA: Structural and functional kinship with the glucocorticoid receptor. Science 237:268–275, 1987

    Google Scholar 

  24. Gunning P, Ponte P, Okayama H, Engel J, Blau H, Kedes L: Isolation and characterizat ion of full-length cDNA clones for human a-, b-, and g-actin mRNAs: Skeletal but not cytoplasmic actins have an amino-terminal cysteine that is subsequently removed. Mol Cell Biol 3:787–795, 1983

    Google Scholar 

  25. Fukushima K, Masuda T, Ohtani H, Sasaki I, Funayama Y, Matsuno S, et al: Immunohistochemical characterization, distribution and ultrastructure of lymphocytes bearing the gamma/ delta T-cell receptor in the human gut. Virchows Arch (B) 60:7–13, 1991

    Google Scholar 

  26. Bradford MM: A rapid and sensitive me thod for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254, 1976

    Google Scholar 

  27. Tanaka Y, Bush KK, Klauck TM, Higgins PJ: Enhancement of butyrate-induced differentiation of HT-29 human colon carcinoma cells by 1,25-dihydroxyvitamin D3. Biochem Pharmacol 38:3859–3865, 1989

    Google Scholar 

  28. Gum JR, Kam WA, Byrd JC, Hicks JW, Sleisenger MH, Kim YS: Effects of sodium butyrate on human colonic adenocarcinoma cells: Induction of placental-like alkaline phosphatase. J Biol Chem 262:1092–1097, 1987

    Google Scholar 

  29. Chung YS, Song IS, Erickson RH, Sleisenger MH, Kim YS: Effect of growth and sodium butyrate on brush border membrane-associated hydrolases in human colorectal cancer cell lines. Cancer Res 45:2976–2982, 1985

    Google Scholar 

  30. Herz F, Halwer M: Preferential alkaline phosphatase isoenzyme induction by sodium butyrate. Biochem Biophys Acta 499:382–391, 1983

    Google Scholar 

  31. Fukushima K, Sasaki I, Hasegawa H, Takahashi K, Naito H, Funayama Y, et al: Sodium butyrate induced-livertype alkaline phosphatase activity in a small intestinal epithelial cell line, IEC6. Dig Dis Sci 43:1116–1123, 1998

    Google Scholar 

  32. Nordstrom C, Dahiqvist A, Josefson L: Quantitative determination of enzymes in different parts of the villi and crypts of rat small intestine. J Histochem Cytochem 15:713–721, 1967

    Google Scholar 

  33. Bell L, Williams L: Histichemical demonstration of alkaline phosphatase in human large intestine, normal and diseased. Histochemistry 60:84–89, 1979

    Google Scholar 

  34. Binder HJ, Sandle GI: Electrolyte transport in the mammalian colon. In Physiology of the Gastrointestinal Tract. LR Johnson (ed). New York, Raven Press, 1994, pp 2133–2171

    Google Scholar 

  35. Weyer A, Huott P, Liu W, McRoberts JA, Dharmsathaphorn K: Chloride secretory me chanism induced by prostaglandin E1 in a colonic epithelial cell line. J Clin Invest 76:1828–1836, 1985

    Google Scholar 

  36. Dharmsathaphorn K, Pandol SJ: Mechanism of chloride secretion induced by carbachol in a colonic epithelial cell line. J Clin Invest 77:348–354, 1987

    Google Scholar 

  37. Wahurst G, Higgs NB, Tonge A, Turnberg LA: Stimulatory and inhibitory actions of carbachol on chloride secretory responses in human colonic cell line T84. Am J Physiol 261:G220-G228, 1991

    Google Scholar 

  38. Scheppach W, Bartram P, Richter A, Richter F, Liepold H, Dusel G, et al: Effect of short chain fatty acids on the human Colonic mucosa in vitro. J Parenter Enter Nutr 16:43–48, 1992

    Google Scholar 

  39. Roediger WEW: The colonic epithelium in ulcerative colitis: An energy-deficiency disease? Lancet 2:712–715, 1980

    Google Scholar 

  40. Kvietys PR, Granger DN: Effect of volatile fatty acids on blood flow and oxygen uptake by the dog colon. Gastroenterology 80:962–969, 1981

    Google Scholar 

  41. Fukushima K, Sasano H, Sasaki I, Nagura H, Funayama Y, Matsuno S: Increased expression of mineralocorticoid receptor in human ileum after total colectomy: Immunohistochemical and immunoblotting studies. Tohoku J Exp Med 173:383–390, 1994

    Google Scholar 

  42. Nasmyth DG, Godwin PGR, Dixon MF, Williams NS, Johnston D: Ileal ecology after pouch-anal anastomosis or ileostomy: A study of mucosal morphology, fecal bacteriology, fecal volatile fatty acids, and their interrelationship. Gastroenterology 96:817–824, 1989

    Google Scholar 

  43. Ambroze WL, Pemberton JH, Phillips SF, Bell AM, Haddad AC: Fecal short-chain fatty acid concentrations and effect on ileal pouch function. Dis Colon Rectum 36:235–239, 1993

    Google Scholar 

  44. Koyama K, Sasaki I, Unno M, Funayama Y, Naito H, Fukushima K, et al: Activation of electrogenic Na absorption is critical for adaptation of rats after total colectomy. Gastroenterology 110:A339, 1996

    Google Scholar 

  45. Gallagher ND, Harrison DD, Skyring AP: Fluid and electrolyte disturbances in patients with long-established ileostomies. Gut 3:219–223, 1962

    Google Scholar 

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Authors
  1. Kouhei Fukushima
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  2. Iwao Sasaki
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  3. Shun Sato
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  4. Hironobu Sasano
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  5. Z. Krozowski
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  6. Seiki Matsuno
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Fukushima, K., Sasaki, I., Sato, S. et al. Induction of Mineralocorticoid Receptor by Sodium Butyrate in Small Intestinal (IEC6) and Colonic (T84) Epithelial Cell Lines. Dig Dis Sci 44, 1571–1578 (1999). https://doi.org/10.1023/A:1026606809171

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  • DOI: https://doi.org/10.1023/A:1026606809171

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