Clonidine enhances type-2 cationic amino acid transporter transcription in endotoxin-activated murine macrophages

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Abstract

Background: We sought to evaluate the effects of clonidine on type-2 cationic amino acid transporter (CAT-2) transcription in endotoxin-activated murine macrophages. Methods: To determine the effects of clonidine on CAT-2 transcription, confluent murine macrophages (RAW264.7 cells) were treated with 1x phosphate buffered saline, clonidine (1000μM), lipopolysaccharide (LPS, 100ng/mL), or LPS plus clonidine (10, 100, or 1000μM). After reacting with LPS for 18 hours or a comparable duration in groups without LPS, cell cultures were harvested and the CAT-2 mRNA concentration was assayed. To determine the stability of CAT-2 mRNA, confluent macrophages were treated with LPS or LPS plus clonidine (100μM). After reacting with LPS for 6 hours, CAT-2 transcription was terminated and the stability of CAT-2 mRNA was determined. Results: The CAT-2 mRNA concentration of cell cultures receiving LPS plus clonidine (100μM) or LPS plus clonidine (1000μM) were significantly higher than that of the cell cultures receiving LPS alone, whereas the CAT-2 mRNA concentrations of cell cultures receiving LPS plus clonidine (10μM) was comparable to that of cell cultures receiving LPS alone. The data indicated that clonidine significantly enhanced LPS-induced CAT-2 transcription. The estimated half-life of CAT-2 mRNA of cell cultures receiving LPS was similar to that of cell cultures receiving LPS plus clonidine. These results indicated that clonidine did not affect CAT-2 mRNA stability. Conclusion: Clonidine enhances CAT-2 transcription in endotoxin-activated murine macrophages.

Original languageEnglish
Pages (from-to)118-123
Number of pages6
JournalActa Anaesthesiologica Taiwanica
Volume46
Issue number3
DOIs
Publication statusPublished - Sep 2008

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Cationic Amino Acid Transporter 2
Clonidine
Endotoxins
Macrophages
Cell Culture Techniques
Messenger RNA
RNA Stability

Keywords

  • Cationic amino acid transporter 2
  • Clonidine
  • Macrophages
  • Tipopolysaccharides

ASJC Scopus subject areas

  • Anesthesiology and Pain Medicine

Cite this

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title = "Clonidine enhances type-2 cationic amino acid transporter transcription in endotoxin-activated murine macrophages",
abstract = "Background: We sought to evaluate the effects of clonidine on type-2 cationic amino acid transporter (CAT-2) transcription in endotoxin-activated murine macrophages. Methods: To determine the effects of clonidine on CAT-2 transcription, confluent murine macrophages (RAW264.7 cells) were treated with 1x phosphate buffered saline, clonidine (1000μM), lipopolysaccharide (LPS, 100ng/mL), or LPS plus clonidine (10, 100, or 1000μM). After reacting with LPS for 18 hours or a comparable duration in groups without LPS, cell cultures were harvested and the CAT-2 mRNA concentration was assayed. To determine the stability of CAT-2 mRNA, confluent macrophages were treated with LPS or LPS plus clonidine (100μM). After reacting with LPS for 6 hours, CAT-2 transcription was terminated and the stability of CAT-2 mRNA was determined. Results: The CAT-2 mRNA concentration of cell cultures receiving LPS plus clonidine (100μM) or LPS plus clonidine (1000μM) were significantly higher than that of the cell cultures receiving LPS alone, whereas the CAT-2 mRNA concentrations of cell cultures receiving LPS plus clonidine (10μM) was comparable to that of cell cultures receiving LPS alone. The data indicated that clonidine significantly enhanced LPS-induced CAT-2 transcription. The estimated half-life of CAT-2 mRNA of cell cultures receiving LPS was similar to that of cell cultures receiving LPS plus clonidine. These results indicated that clonidine did not affect CAT-2 mRNA stability. Conclusion: Clonidine enhances CAT-2 transcription in endotoxin-activated murine macrophages.",
keywords = "Cationic amino acid transporter 2, Clonidine, Macrophages, Tipopolysaccharides",
author = "Lai, {Yen Chun} and Pei-Shan Tsai and Huang, {Chun Jen}",
year = "2008",
month = "9",
doi = "10.1016/S1875-4597(08)60005-3",
language = "English",
volume = "46",
pages = "118--123",
journal = "Asian Journal of Anesthesiology",
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TY - JOUR

T1 - Clonidine enhances type-2 cationic amino acid transporter transcription in endotoxin-activated murine macrophages

AU - Lai, Yen Chun

AU - Tsai, Pei-Shan

AU - Huang, Chun Jen

PY - 2008/9

Y1 - 2008/9

N2 - Background: We sought to evaluate the effects of clonidine on type-2 cationic amino acid transporter (CAT-2) transcription in endotoxin-activated murine macrophages. Methods: To determine the effects of clonidine on CAT-2 transcription, confluent murine macrophages (RAW264.7 cells) were treated with 1x phosphate buffered saline, clonidine (1000μM), lipopolysaccharide (LPS, 100ng/mL), or LPS plus clonidine (10, 100, or 1000μM). After reacting with LPS for 18 hours or a comparable duration in groups without LPS, cell cultures were harvested and the CAT-2 mRNA concentration was assayed. To determine the stability of CAT-2 mRNA, confluent macrophages were treated with LPS or LPS plus clonidine (100μM). After reacting with LPS for 6 hours, CAT-2 transcription was terminated and the stability of CAT-2 mRNA was determined. Results: The CAT-2 mRNA concentration of cell cultures receiving LPS plus clonidine (100μM) or LPS plus clonidine (1000μM) were significantly higher than that of the cell cultures receiving LPS alone, whereas the CAT-2 mRNA concentrations of cell cultures receiving LPS plus clonidine (10μM) was comparable to that of cell cultures receiving LPS alone. The data indicated that clonidine significantly enhanced LPS-induced CAT-2 transcription. The estimated half-life of CAT-2 mRNA of cell cultures receiving LPS was similar to that of cell cultures receiving LPS plus clonidine. These results indicated that clonidine did not affect CAT-2 mRNA stability. Conclusion: Clonidine enhances CAT-2 transcription in endotoxin-activated murine macrophages.

AB - Background: We sought to evaluate the effects of clonidine on type-2 cationic amino acid transporter (CAT-2) transcription in endotoxin-activated murine macrophages. Methods: To determine the effects of clonidine on CAT-2 transcription, confluent murine macrophages (RAW264.7 cells) were treated with 1x phosphate buffered saline, clonidine (1000μM), lipopolysaccharide (LPS, 100ng/mL), or LPS plus clonidine (10, 100, or 1000μM). After reacting with LPS for 18 hours or a comparable duration in groups without LPS, cell cultures were harvested and the CAT-2 mRNA concentration was assayed. To determine the stability of CAT-2 mRNA, confluent macrophages were treated with LPS or LPS plus clonidine (100μM). After reacting with LPS for 6 hours, CAT-2 transcription was terminated and the stability of CAT-2 mRNA was determined. Results: The CAT-2 mRNA concentration of cell cultures receiving LPS plus clonidine (100μM) or LPS plus clonidine (1000μM) were significantly higher than that of the cell cultures receiving LPS alone, whereas the CAT-2 mRNA concentrations of cell cultures receiving LPS plus clonidine (10μM) was comparable to that of cell cultures receiving LPS alone. The data indicated that clonidine significantly enhanced LPS-induced CAT-2 transcription. The estimated half-life of CAT-2 mRNA of cell cultures receiving LPS was similar to that of cell cultures receiving LPS plus clonidine. These results indicated that clonidine did not affect CAT-2 mRNA stability. Conclusion: Clonidine enhances CAT-2 transcription in endotoxin-activated murine macrophages.

KW - Cationic amino acid transporter 2

KW - Clonidine

KW - Macrophages

KW - Tipopolysaccharides

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