Ketamine inhibits agonist-induced cAMP accumulation increase in human airway smooth muscle cells

Gary E. Hill, Jodi L. Anderson, Charles W. Whitten

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Purpose: Cyclic 3',5' adenosine monophosphate (cAMP) is a second messenger of the beta adrenergic receptor (BAR). Ketamine causes an increase in the intracellular accumulation of cAMP in several non-human tissue preparations. A 'species effect' may explain the differing results of ketamine on βAR mediated responses, thus reports of a ketamine-induced increase in cAMP in other species may not be applicable to humans. Methods: The effect of ketamine (10-3, 10-4, or 10-5 M) pretreatment (60 and 120 min) on isoproterenol [ISO, a beta adrenergic receptor (AR) agonist] or forskolin [FSK, an activator of adenylylcyclase (AC)]-induced intracellular accumulation of cAMP in a human airway smooth muscle (tracheal) cell line (HASM) was evaluated. In an in vitro HASM culture, cells with or without pretreatment were labeled with [3H]adenine to produce [3H]ATP, and following stimulation with ISO or FSK to convert the [3H]ATP to [3H]cAMP, the intracellular accumulation of [3H]cAMP was measured by sequential chromatography over Dowex and alumina columns. Results: Pretreatment of the HASM cells with ketamine (10-3 and 10-4 M) caused a reduction (P < 0.05, when compared to untreated cells) in ISO-induced cAMP accumulation, but did not effect cAMP accumulation following FSK stimulation. This effect of ketamine was greater at 120 min of pretreatment than at 60 min (10-3 M ketamine only)(P < 0.05). No effect was found at either time period following pretreatment of the HASM cells with ketamine 10-5M. Conclusions: These results demonstrate that pretreatment of the HASM cells with ketamine reduces ISO-induced cAMP accumulation. Since only ISO-induced cAMP was effected by ketamine, these data suggest that ketamine inhibits production of CAMP proximal to AC in the cAMP production pathway. These results also demonstrate that a mechanism other than that involving the βAR and intracellular cAMP accumulation is responsible for the ketamine induced bronchodilation in humans.

Original languageEnglish (US)
Pages (from-to)1172-1177
Number of pages6
JournalCanadian Journal of Anaesthesia
Volume46
Issue number12
StatePublished - 1999

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Ketamine
Cyclic AMP
Smooth Muscle Myocytes
Adrenergic Receptors
Adenosine Triphosphate
Adrenergic beta-Agonists
Aluminum Oxide
Receptors, Adrenergic, beta
Second Messenger Systems
Adenine
Colforsin
Isoproterenol
Chromatography
Cell Culture Techniques

ASJC Scopus subject areas

  • Anesthesiology and Pain Medicine

Cite this

Ketamine inhibits agonist-induced cAMP accumulation increase in human airway smooth muscle cells. / Hill, Gary E.; Anderson, Jodi L.; Whitten, Charles W.

In: Canadian Journal of Anaesthesia, Vol. 46, No. 12, 1999, p. 1172-1177.

Research output: Contribution to journalArticle

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abstract = "Purpose: Cyclic 3',5' adenosine monophosphate (cAMP) is a second messenger of the beta adrenergic receptor (BAR). Ketamine causes an increase in the intracellular accumulation of cAMP in several non-human tissue preparations. A 'species effect' may explain the differing results of ketamine on βAR mediated responses, thus reports of a ketamine-induced increase in cAMP in other species may not be applicable to humans. Methods: The effect of ketamine (10-3, 10-4, or 10-5 M) pretreatment (60 and 120 min) on isoproterenol [ISO, a beta adrenergic receptor (AR) agonist] or forskolin [FSK, an activator of adenylylcyclase (AC)]-induced intracellular accumulation of cAMP in a human airway smooth muscle (tracheal) cell line (HASM) was evaluated. In an in vitro HASM culture, cells with or without pretreatment were labeled with [3H]adenine to produce [3H]ATP, and following stimulation with ISO or FSK to convert the [3H]ATP to [3H]cAMP, the intracellular accumulation of [3H]cAMP was measured by sequential chromatography over Dowex and alumina columns. Results: Pretreatment of the HASM cells with ketamine (10-3 and 10-4 M) caused a reduction (P < 0.05, when compared to untreated cells) in ISO-induced cAMP accumulation, but did not effect cAMP accumulation following FSK stimulation. This effect of ketamine was greater at 120 min of pretreatment than at 60 min (10-3 M ketamine only)(P < 0.05). No effect was found at either time period following pretreatment of the HASM cells with ketamine 10-5M. Conclusions: These results demonstrate that pretreatment of the HASM cells with ketamine reduces ISO-induced cAMP accumulation. Since only ISO-induced cAMP was effected by ketamine, these data suggest that ketamine inhibits production of CAMP proximal to AC in the cAMP production pathway. These results also demonstrate that a mechanism other than that involving the βAR and intracellular cAMP accumulation is responsible for the ketamine induced bronchodilation in humans.",
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