The direct effect of levobupivacaine in isolated rat aorta involves lipoxygenase pathway activation and endothelial nitric oxide release

Yun Suk Choi, Young Seok Jeong, Seong Ho Ok, Il Woo Shin, Seung Hwa Lee, Jae-Yong Park, Eun Mi Hwang, Young Sool Hah, Ju Tae Sohn

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Background: Levobupivacaine is a long-acting local anesthetic with a clinical profile similar to that of racemic bupivacaine but with a greater margin of safety. Levobupivacaine produces dose-dependent vasoconstriction in vivo. Our goal in this in vitro study was to investigate the role of pathways involved in arachidonic acid metabolism in the levobupivacaine-induced contraction of isolated rat aorta and to determine which endothelium-derived vasodilators are involved in the modulation of levobupivacaine-induced contraction. Methods: Rat thoracic aortic rings were isolated and suspended for isometric tension recording. Cumulative levobupivacaine dose-response curves over a range of 10 -6 to 3 × 10 -4 M were constructed in 1) aortic rings with no drug pretreatment; 2) endothelium-denuded rings pretreated with quinacrine dihydrochloride (nonspecific phospholipase A 2 inhibitor: 2 × 10 -5 , 4 × 10 -5 M), nordihydroguaiaretic acid (NDGA) (lipoxygenase inhibitor: 10 -5 , 3 × 10 M), indomethacin (nonspecific cyclooxygenase inhibitor: 10 M), AA-861 (5-lipoxygenase inhibitor: 10 -5 , 5 × 10 -5 M), fluconazole (cytochrome P450 epoxygenase inhibitor: 10 -5 M), verapamil (10 M), or calcium-free solution; and 3) endothelium-intact rings pretreated with N-nitro-l-arginine methyl ester (L-NAME) (nitric oxide synthase inhibitor: 5 × 10 -5 M), indomethacin, or fluconazole. Levobupivacaine-induced contractile response at each concentration (10 -4 , 3 × 10 -4 M) was assessed in endothelium-denuded rings. Dose-response curves for potassium chloride in endothelium-denuded rings were generated in the presence or absence of NDGA and AA-861. Intracellular Ca 2+ levels were monitored by Ca 2+ image analysis using Fluo-4 fluorescence in vascular smooth muscle cells treated with levobupivacaine alone or AA-861 plus levobupivacaine. Results: Levobupivacaine produced a tonic contraction in isolated rat aorta rings; this response was maximal at 10 -4 M levobupivacaine and gradually attenuated at 3 × 10 M levobupivacaine. Levobupivacaine-induced contractions of endothelium-denuded rings were larger than those of endothelium-intact rings. Levobupivacaine- induced contraction of endothelium-denuded rings was attenuated by quinacrine dihydrochloride, NDGA, AA-861, verapamil, and calcium-free solution and, to a lesser extent, by indomethacin. L-NAME enhanced levobupivacaine-induced contraction of endothelium-intact rings and indomethacin slightly attenuated this contraction. NDGA and AA-861 attenuated the potassium chloride-induced contraction. AA-861 attenuated the levobupivacaine-induced intracellular calcium increase in vascular smooth muscle cells. Conclusions: Our data indicate that levobupivacaine-induced contraction of rat aortic smooth muscle is mediated mainly by activation of the lipoxygenase pathway and in part by activation of the cyclooxygenase pathway. In addition, activation of the lipoxygenase pathway seems to facilitate calcium influx via L-type calcium channels. Endothelial nitric oxide attenuates levobupivacaine-induced contraction.

Original languageEnglish
Pages (from-to)341-349
Number of pages9
JournalAnesthesia and Analgesia
Volume110
Issue number2
DOIs
Publication statusPublished - 2010 Jan 1
Externally publishedYes

Fingerprint

Lipoxygenase
Aorta
Nitric Oxide
Endothelium
Masoprocol
Indomethacin
Calcium
Quinacrine
Lipoxygenase Inhibitors
levobupivacaine
Potassium Chloride
Fluconazole
NG-Nitroarginine Methyl Ester
Verapamil
Vascular Smooth Muscle
Smooth Muscle Myocytes
L-Type Calcium Channels
Cyclooxygenase Inhibitors
Phospholipases A
Bupivacaine

ASJC Scopus subject areas

  • Anesthesiology and Pain Medicine

Cite this

The direct effect of levobupivacaine in isolated rat aorta involves lipoxygenase pathway activation and endothelial nitric oxide release. / Choi, Yun Suk; Jeong, Young Seok; Ok, Seong Ho; Shin, Il Woo; Lee, Seung Hwa; Park, Jae-Yong; Hwang, Eun Mi; Hah, Young Sool; Sohn, Ju Tae.

In: Anesthesia and Analgesia, Vol. 110, No. 2, 01.01.2010, p. 341-349.

Research output: Contribution to journalArticle

Choi, Yun Suk ; Jeong, Young Seok ; Ok, Seong Ho ; Shin, Il Woo ; Lee, Seung Hwa ; Park, Jae-Yong ; Hwang, Eun Mi ; Hah, Young Sool ; Sohn, Ju Tae. / The direct effect of levobupivacaine in isolated rat aorta involves lipoxygenase pathway activation and endothelial nitric oxide release. In: Anesthesia and Analgesia. 2010 ; Vol. 110, No. 2. pp. 341-349.
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TY - JOUR

T1 - The direct effect of levobupivacaine in isolated rat aorta involves lipoxygenase pathway activation and endothelial nitric oxide release

AU - Choi, Yun Suk

AU - Jeong, Young Seok

AU - Ok, Seong Ho

AU - Shin, Il Woo

AU - Lee, Seung Hwa

AU - Park, Jae-Yong

AU - Hwang, Eun Mi

AU - Hah, Young Sool

AU - Sohn, Ju Tae

PY - 2010/1/1

Y1 - 2010/1/1

N2 - Background: Levobupivacaine is a long-acting local anesthetic with a clinical profile similar to that of racemic bupivacaine but with a greater margin of safety. Levobupivacaine produces dose-dependent vasoconstriction in vivo. Our goal in this in vitro study was to investigate the role of pathways involved in arachidonic acid metabolism in the levobupivacaine-induced contraction of isolated rat aorta and to determine which endothelium-derived vasodilators are involved in the modulation of levobupivacaine-induced contraction. Methods: Rat thoracic aortic rings were isolated and suspended for isometric tension recording. Cumulative levobupivacaine dose-response curves over a range of 10 -6 to 3 × 10 -4 M were constructed in 1) aortic rings with no drug pretreatment; 2) endothelium-denuded rings pretreated with quinacrine dihydrochloride (nonspecific phospholipase A 2 inhibitor: 2 × 10 -5 , 4 × 10 -5 M), nordihydroguaiaretic acid (NDGA) (lipoxygenase inhibitor: 10 -5 , 3 × 10 M), indomethacin (nonspecific cyclooxygenase inhibitor: 10 M), AA-861 (5-lipoxygenase inhibitor: 10 -5 , 5 × 10 -5 M), fluconazole (cytochrome P450 epoxygenase inhibitor: 10 -5 M), verapamil (10 M), or calcium-free solution; and 3) endothelium-intact rings pretreated with N-nitro-l-arginine methyl ester (L-NAME) (nitric oxide synthase inhibitor: 5 × 10 -5 M), indomethacin, or fluconazole. Levobupivacaine-induced contractile response at each concentration (10 -4 , 3 × 10 -4 M) was assessed in endothelium-denuded rings. Dose-response curves for potassium chloride in endothelium-denuded rings were generated in the presence or absence of NDGA and AA-861. Intracellular Ca 2+ levels were monitored by Ca 2+ image analysis using Fluo-4 fluorescence in vascular smooth muscle cells treated with levobupivacaine alone or AA-861 plus levobupivacaine. Results: Levobupivacaine produced a tonic contraction in isolated rat aorta rings; this response was maximal at 10 -4 M levobupivacaine and gradually attenuated at 3 × 10 M levobupivacaine. Levobupivacaine-induced contractions of endothelium-denuded rings were larger than those of endothelium-intact rings. Levobupivacaine- induced contraction of endothelium-denuded rings was attenuated by quinacrine dihydrochloride, NDGA, AA-861, verapamil, and calcium-free solution and, to a lesser extent, by indomethacin. L-NAME enhanced levobupivacaine-induced contraction of endothelium-intact rings and indomethacin slightly attenuated this contraction. NDGA and AA-861 attenuated the potassium chloride-induced contraction. AA-861 attenuated the levobupivacaine-induced intracellular calcium increase in vascular smooth muscle cells. Conclusions: Our data indicate that levobupivacaine-induced contraction of rat aortic smooth muscle is mediated mainly by activation of the lipoxygenase pathway and in part by activation of the cyclooxygenase pathway. In addition, activation of the lipoxygenase pathway seems to facilitate calcium influx via L-type calcium channels. Endothelial nitric oxide attenuates levobupivacaine-induced contraction.

AB - Background: Levobupivacaine is a long-acting local anesthetic with a clinical profile similar to that of racemic bupivacaine but with a greater margin of safety. Levobupivacaine produces dose-dependent vasoconstriction in vivo. Our goal in this in vitro study was to investigate the role of pathways involved in arachidonic acid metabolism in the levobupivacaine-induced contraction of isolated rat aorta and to determine which endothelium-derived vasodilators are involved in the modulation of levobupivacaine-induced contraction. Methods: Rat thoracic aortic rings were isolated and suspended for isometric tension recording. Cumulative levobupivacaine dose-response curves over a range of 10 -6 to 3 × 10 -4 M were constructed in 1) aortic rings with no drug pretreatment; 2) endothelium-denuded rings pretreated with quinacrine dihydrochloride (nonspecific phospholipase A 2 inhibitor: 2 × 10 -5 , 4 × 10 -5 M), nordihydroguaiaretic acid (NDGA) (lipoxygenase inhibitor: 10 -5 , 3 × 10 M), indomethacin (nonspecific cyclooxygenase inhibitor: 10 M), AA-861 (5-lipoxygenase inhibitor: 10 -5 , 5 × 10 -5 M), fluconazole (cytochrome P450 epoxygenase inhibitor: 10 -5 M), verapamil (10 M), or calcium-free solution; and 3) endothelium-intact rings pretreated with N-nitro-l-arginine methyl ester (L-NAME) (nitric oxide synthase inhibitor: 5 × 10 -5 M), indomethacin, or fluconazole. Levobupivacaine-induced contractile response at each concentration (10 -4 , 3 × 10 -4 M) was assessed in endothelium-denuded rings. Dose-response curves for potassium chloride in endothelium-denuded rings were generated in the presence or absence of NDGA and AA-861. Intracellular Ca 2+ levels were monitored by Ca 2+ image analysis using Fluo-4 fluorescence in vascular smooth muscle cells treated with levobupivacaine alone or AA-861 plus levobupivacaine. Results: Levobupivacaine produced a tonic contraction in isolated rat aorta rings; this response was maximal at 10 -4 M levobupivacaine and gradually attenuated at 3 × 10 M levobupivacaine. Levobupivacaine-induced contractions of endothelium-denuded rings were larger than those of endothelium-intact rings. Levobupivacaine- induced contraction of endothelium-denuded rings was attenuated by quinacrine dihydrochloride, NDGA, AA-861, verapamil, and calcium-free solution and, to a lesser extent, by indomethacin. L-NAME enhanced levobupivacaine-induced contraction of endothelium-intact rings and indomethacin slightly attenuated this contraction. NDGA and AA-861 attenuated the potassium chloride-induced contraction. AA-861 attenuated the levobupivacaine-induced intracellular calcium increase in vascular smooth muscle cells. Conclusions: Our data indicate that levobupivacaine-induced contraction of rat aortic smooth muscle is mediated mainly by activation of the lipoxygenase pathway and in part by activation of the cyclooxygenase pathway. In addition, activation of the lipoxygenase pathway seems to facilitate calcium influx via L-type calcium channels. Endothelial nitric oxide attenuates levobupivacaine-induced contraction.

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