Regulation and expression of endothelin-1 (ET-1) and ET-receptors in rat epithelial cells of renal and intestinal origin

Femke M van de Water; Frans G M Russel; R. Masereeuw

doi:10.1016/j.phrs.2006.08.006

Regulation and expression of endothelin-1 (ET-1) and ET-receptors in rat epithelial cells of renal and intestinal origin

Femke M van de Water, Frans G M Russel, R. Masereeuw

Pharmacology

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

The hormone endothelin-1 (ET-1) is involved in many functions of the kidney and intestine. In addition to its vasoactive and proliferative effects, ET-1 is involved in the maintenance of water and salt balance, and in drug excretion by influencing the activity of different transporters in the epithelial cells of these two organs. To study ET-1 function and its role in pathophysiological processes in epithelial cells in vitro, we investigated ET-1 and ET-receptor expression and inducibility of ET-1 excretion by cytokines in three rat cell lines of intestinal (IEC-6) and renal (NRK-52E and GERP) origin. Immunocytochemistry showed that all three cell lines express ET-1 and the ET-A and ET-B receptor. ET-1 was expressed intracellularly, and also the ET-A receptor showed a punctate intracellular staining pattern. The ET-B receptor was localized in the membrane, which was confirmed by Western blot analysis. Real-time RT-PCR and ELISA showed that exposure of IEC-6 cells to the cytokines, interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNFalpha), induced ET-1 mRNA expression and excretion, while IL-2 was ineffective. In NRK-52E cells, IL-1beta and TNFalpha induced ET-1 excretion as well. In GERP cells, adequate measurement of cytokine effects on ET-1 excretion was not possible, since ET-1 excretion under non-stimulated conditions was around the lowest level of detection. In conclusion, we showed ET-1 and ET-receptor expression, and inducibility of ET-1 by cytokines in IEC-6, NRK-52E, and GERP cells. These rat intestinal and renal cell lines appear to be suitable for further characterisation of ET-1 function and its role in pathophysiological processes in epithelial cells.

Original language	English
Pages (from-to)	429-35
Number of pages	7
Journal	Pharmacological Research
Volume	54
Issue number	6
DOIs	https://doi.org/10.1016/j.phrs.2006.08.006
Publication status	Published - Dec 2006

Keywords

Animals
Blotting, Western
Cell Line
Cytokines
Endothelin-1
Epithelial Cells
Fluorescent Antibody Technique
Immunohistochemistry
Intestines
Kidney
RNA, Messenger
Rats
Receptors, Endothelin
Reverse Transcriptase Polymerase Chain Reaction

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10.1016/j.phrs.2006.08.006

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@article{84bbc93e9f46431181c53f97027c0df9,

title = "Regulation and expression of endothelin-1 (ET-1) and ET-receptors in rat epithelial cells of renal and intestinal origin",

abstract = "The hormone endothelin-1 (ET-1) is involved in many functions of the kidney and intestine. In addition to its vasoactive and proliferative effects, ET-1 is involved in the maintenance of water and salt balance, and in drug excretion by influencing the activity of different transporters in the epithelial cells of these two organs. To study ET-1 function and its role in pathophysiological processes in epithelial cells in vitro, we investigated ET-1 and ET-receptor expression and inducibility of ET-1 excretion by cytokines in three rat cell lines of intestinal (IEC-6) and renal (NRK-52E and GERP) origin. Immunocytochemistry showed that all three cell lines express ET-1 and the ET-A and ET-B receptor. ET-1 was expressed intracellularly, and also the ET-A receptor showed a punctate intracellular staining pattern. The ET-B receptor was localized in the membrane, which was confirmed by Western blot analysis. Real-time RT-PCR and ELISA showed that exposure of IEC-6 cells to the cytokines, interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNFalpha), induced ET-1 mRNA expression and excretion, while IL-2 was ineffective. In NRK-52E cells, IL-1beta and TNFalpha induced ET-1 excretion as well. In GERP cells, adequate measurement of cytokine effects on ET-1 excretion was not possible, since ET-1 excretion under non-stimulated conditions was around the lowest level of detection. In conclusion, we showed ET-1 and ET-receptor expression, and inducibility of ET-1 by cytokines in IEC-6, NRK-52E, and GERP cells. These rat intestinal and renal cell lines appear to be suitable for further characterisation of ET-1 function and its role in pathophysiological processes in epithelial cells.",

keywords = "Animals, Blotting, Western, Cell Line, Cytokines, Endothelin-1, Epithelial Cells, Fluorescent Antibody Technique, Immunohistochemistry, Intestines, Kidney, RNA, Messenger, Rats, Receptors, Endothelin, Reverse Transcriptase Polymerase Chain Reaction",

author = "{van de Water}, {Femke M} and Russel, {Frans G M} and R. Masereeuw",

year = "2006",

month = dec,

doi = "10.1016/j.phrs.2006.08.006",

language = "English",

volume = "54",

pages = "429--35",

journal = "Pharmacological Research",

issn = "1043-6618",

publisher = "Academic Press",

number = "6",

}

TY - JOUR

T1 - Regulation and expression of endothelin-1 (ET-1) and ET-receptors in rat epithelial cells of renal and intestinal origin

AU - van de Water, Femke M

AU - Russel, Frans G M

AU - Masereeuw, R.

PY - 2006/12

Y1 - 2006/12

N2 - The hormone endothelin-1 (ET-1) is involved in many functions of the kidney and intestine. In addition to its vasoactive and proliferative effects, ET-1 is involved in the maintenance of water and salt balance, and in drug excretion by influencing the activity of different transporters in the epithelial cells of these two organs. To study ET-1 function and its role in pathophysiological processes in epithelial cells in vitro, we investigated ET-1 and ET-receptor expression and inducibility of ET-1 excretion by cytokines in three rat cell lines of intestinal (IEC-6) and renal (NRK-52E and GERP) origin. Immunocytochemistry showed that all three cell lines express ET-1 and the ET-A and ET-B receptor. ET-1 was expressed intracellularly, and also the ET-A receptor showed a punctate intracellular staining pattern. The ET-B receptor was localized in the membrane, which was confirmed by Western blot analysis. Real-time RT-PCR and ELISA showed that exposure of IEC-6 cells to the cytokines, interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNFalpha), induced ET-1 mRNA expression and excretion, while IL-2 was ineffective. In NRK-52E cells, IL-1beta and TNFalpha induced ET-1 excretion as well. In GERP cells, adequate measurement of cytokine effects on ET-1 excretion was not possible, since ET-1 excretion under non-stimulated conditions was around the lowest level of detection. In conclusion, we showed ET-1 and ET-receptor expression, and inducibility of ET-1 by cytokines in IEC-6, NRK-52E, and GERP cells. These rat intestinal and renal cell lines appear to be suitable for further characterisation of ET-1 function and its role in pathophysiological processes in epithelial cells.

AB - The hormone endothelin-1 (ET-1) is involved in many functions of the kidney and intestine. In addition to its vasoactive and proliferative effects, ET-1 is involved in the maintenance of water and salt balance, and in drug excretion by influencing the activity of different transporters in the epithelial cells of these two organs. To study ET-1 function and its role in pathophysiological processes in epithelial cells in vitro, we investigated ET-1 and ET-receptor expression and inducibility of ET-1 excretion by cytokines in three rat cell lines of intestinal (IEC-6) and renal (NRK-52E and GERP) origin. Immunocytochemistry showed that all three cell lines express ET-1 and the ET-A and ET-B receptor. ET-1 was expressed intracellularly, and also the ET-A receptor showed a punctate intracellular staining pattern. The ET-B receptor was localized in the membrane, which was confirmed by Western blot analysis. Real-time RT-PCR and ELISA showed that exposure of IEC-6 cells to the cytokines, interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNFalpha), induced ET-1 mRNA expression and excretion, while IL-2 was ineffective. In NRK-52E cells, IL-1beta and TNFalpha induced ET-1 excretion as well. In GERP cells, adequate measurement of cytokine effects on ET-1 excretion was not possible, since ET-1 excretion under non-stimulated conditions was around the lowest level of detection. In conclusion, we showed ET-1 and ET-receptor expression, and inducibility of ET-1 by cytokines in IEC-6, NRK-52E, and GERP cells. These rat intestinal and renal cell lines appear to be suitable for further characterisation of ET-1 function and its role in pathophysiological processes in epithelial cells.

KW - Animals

KW - Blotting, Western

KW - Cell Line

KW - Cytokines

KW - Endothelin-1

KW - Epithelial Cells

KW - Fluorescent Antibody Technique

KW - Immunohistochemistry

KW - Intestines

KW - Kidney

KW - RNA, Messenger

KW - Rats

KW - Receptors, Endothelin

KW - Reverse Transcriptase Polymerase Chain Reaction

U2 - 10.1016/j.phrs.2006.08.006

DO - 10.1016/j.phrs.2006.08.006

M3 - Article

C2 - 17000116

SN - 1043-6618

VL - 54

SP - 429

EP - 435

JO - Pharmacological Research

JF - Pharmacological Research

IS - 6

ER -

Regulation and expression of endothelin-1 (ET-1) and ET-receptors in rat epithelial cells of renal and intestinal origin

Abstract

Keywords

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