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Propofol and aminophylline antagonize each other during the mobilization of intracellular calcium in human umbilical vein endothelial cells.

Son HJ, Lim YC, Ha KS, Kang SS, Cheong IY, Lee SJ, Park SW, Hwang BM - J. Korean Med. Sci. (2010)

Bottom Line: The results were expressed as relative fluorescence intensity and fold stimulation.Propofol further prevented the aminophylline-induced increase of intracellular calcium at clinically relevant concentrations.However, aminophylline reversed the inhibitory effect of propofol on the elevation of intracellular calcium by lysophosphatidic acid.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesiology and Pain Medicine, Kangwon National University Medical School, Chuncheon, Korea.

ABSTRACT
This study examined whether propofol and aminophylline affect the mobilization of intracellular calcium in human umbilical vein endothelial cells. Intracellular calcium was measured using laser scanning confocal microscopy. Cultured and serum-starved cells on round coverslips were incubated with propofol or aminophylline for 30 min, and then stimulated with lysophosphatidic acid, propofol and aminophylline. The results were expressed as relative fluorescence intensity and fold stimulation. Propofol decreased the concentration of intracellular calcium, whereas aminophylline caused increased mobilization of intracellular calcium in a concentration-dependent manner. Propofol suppressed the lysophosphatidic acid-induced mobilization of intracellular calcium in a concentration-dependent manner. Propofol further prevented the aminophylline-induced increase of intracellular calcium at clinically relevant concentrations. However, aminophylline reversed the inhibitory effect of propofol on the elevation of intracellular calcium by lysophosphatidic acid. Our results suggest that propofol and aminophylline antagonize each other on the mobilization of intracellular calcium in human umbilical vein endothelial cells at clinically relevant concentrations. Serious consideration should be given to how this interaction affects mobilization of intracellular calcium when these two drugs are used together.

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Levels of mean peak intracellular calcium ([Ca2+]i) generated by lysophosphatidic acid (LPA) or aminophylline treatment after incubation with propofol (10, 30 µM) for 30 min. (A) Levels of mean peak intracellular calcium ([Ca2+]i) generated by lysophosphatidic acid (LPA) or aminophylline treatment after incubation with propofol (10 µM) and for 30 min. LPA only: treatment with LPA (5 µg/mL); P10→LPA: incubation with propofol (10 µM) and treatment with LPA (5 µg/mLl); A100 only: treatment with aminophylline (100 µM); P10→A100: incubation with propofol (10 µM) and treatment with aminophylline (100 µM) Values represent the mean peak intracellular calcium response. Results are expressed as fold-stimulation, determined by comparing relative fluorescence intensities (RFI) before stimulation and expressed as mean±SD from three separate determinations. Each determination represents the mean of at least 10 cells. (B) Levels of mean peak intracellular calcium ([Ca2+]i) generated by lysophosphatidic acid (LPA) or aminophylline treatment after incubation with propofol (30 µM) and for 30 min. LPA only: treatment with LPA (5 µg/mL); P30→LPA: incubation with propofol (30 µM) and treatment with LPA (5 µg/mL); A100 only: treatment with aminophylline (100 µM); P30→A100: incubation with propofol (30 µM) and treatment with aminophylline (100 µM) Values represent the mean peak intracellular calcium response. Results are expressed as fold-stimulation, determined by comparing relative fluorescence intensities (RFI) before stimulation and expressed as mean±SD from three separate determinations. Each determination represents the mean of at least 10 cells.*P<0.05 compared with treatment with LPA (5 µg/mL); †P<0.05 compared with treatment with aminophylline (100 µM).
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Figure 4: Levels of mean peak intracellular calcium ([Ca2+]i) generated by lysophosphatidic acid (LPA) or aminophylline treatment after incubation with propofol (10, 30 µM) for 30 min. (A) Levels of mean peak intracellular calcium ([Ca2+]i) generated by lysophosphatidic acid (LPA) or aminophylline treatment after incubation with propofol (10 µM) and for 30 min. LPA only: treatment with LPA (5 µg/mL); P10→LPA: incubation with propofol (10 µM) and treatment with LPA (5 µg/mLl); A100 only: treatment with aminophylline (100 µM); P10→A100: incubation with propofol (10 µM) and treatment with aminophylline (100 µM) Values represent the mean peak intracellular calcium response. Results are expressed as fold-stimulation, determined by comparing relative fluorescence intensities (RFI) before stimulation and expressed as mean±SD from three separate determinations. Each determination represents the mean of at least 10 cells. (B) Levels of mean peak intracellular calcium ([Ca2+]i) generated by lysophosphatidic acid (LPA) or aminophylline treatment after incubation with propofol (30 µM) and for 30 min. LPA only: treatment with LPA (5 µg/mL); P30→LPA: incubation with propofol (30 µM) and treatment with LPA (5 µg/mL); A100 only: treatment with aminophylline (100 µM); P30→A100: incubation with propofol (30 µM) and treatment with aminophylline (100 µM) Values represent the mean peak intracellular calcium response. Results are expressed as fold-stimulation, determined by comparing relative fluorescence intensities (RFI) before stimulation and expressed as mean±SD from three separate determinations. Each determination represents the mean of at least 10 cells.*P<0.05 compared with treatment with LPA (5 µg/mL); †P<0.05 compared with treatment with aminophylline (100 µM).

Mentions: Aminophylline induced a very rapid, dose-dependent increase in [Ca2+]i (Fig. 3), and propofol (300 µM) treatment decreased the concentration of [Ca2+]i. Aminophylline (1,000 µM) rapidly increased Fluo-4 fluorescence in cells to a maximum three to four-fold higher than the control (Fig. 3). Propofol (10 µM) treatment for 30 min decreased [Ca2+]i induced by LPA (5 µg/mL) and aminophylline (100 µM) (Fig. 4A). Propofol (30 µM) showed similar activity (Fig. 4B). Following incubation with propofol (30 µM) and aminophylline (100; 1,000 µM) for 30 min, the peak level of [Ca2+]i following LPA (5 µg/mL) treatment was higher than propofol (30 µM) only (Fig. 5). Furthermore, HUVECs incubated with propofol (30 µM) and aminophylline (1,000 µM) for 30 min had peak [Ca2+]i levels higher than propofol (30 µM) and aminophylline (100 µM) (Fig. 5).


Propofol and aminophylline antagonize each other during the mobilization of intracellular calcium in human umbilical vein endothelial cells.

Son HJ, Lim YC, Ha KS, Kang SS, Cheong IY, Lee SJ, Park SW, Hwang BM - J. Korean Med. Sci. (2010)

Levels of mean peak intracellular calcium ([Ca2+]i) generated by lysophosphatidic acid (LPA) or aminophylline treatment after incubation with propofol (10, 30 µM) for 30 min. (A) Levels of mean peak intracellular calcium ([Ca2+]i) generated by lysophosphatidic acid (LPA) or aminophylline treatment after incubation with propofol (10 µM) and for 30 min. LPA only: treatment with LPA (5 µg/mL); P10→LPA: incubation with propofol (10 µM) and treatment with LPA (5 µg/mLl); A100 only: treatment with aminophylline (100 µM); P10→A100: incubation with propofol (10 µM) and treatment with aminophylline (100 µM) Values represent the mean peak intracellular calcium response. Results are expressed as fold-stimulation, determined by comparing relative fluorescence intensities (RFI) before stimulation and expressed as mean±SD from three separate determinations. Each determination represents the mean of at least 10 cells. (B) Levels of mean peak intracellular calcium ([Ca2+]i) generated by lysophosphatidic acid (LPA) or aminophylline treatment after incubation with propofol (30 µM) and for 30 min. LPA only: treatment with LPA (5 µg/mL); P30→LPA: incubation with propofol (30 µM) and treatment with LPA (5 µg/mL); A100 only: treatment with aminophylline (100 µM); P30→A100: incubation with propofol (30 µM) and treatment with aminophylline (100 µM) Values represent the mean peak intracellular calcium response. Results are expressed as fold-stimulation, determined by comparing relative fluorescence intensities (RFI) before stimulation and expressed as mean±SD from three separate determinations. Each determination represents the mean of at least 10 cells.*P<0.05 compared with treatment with LPA (5 µg/mL); †P<0.05 compared with treatment with aminophylline (100 µM).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 4: Levels of mean peak intracellular calcium ([Ca2+]i) generated by lysophosphatidic acid (LPA) or aminophylline treatment after incubation with propofol (10, 30 µM) for 30 min. (A) Levels of mean peak intracellular calcium ([Ca2+]i) generated by lysophosphatidic acid (LPA) or aminophylline treatment after incubation with propofol (10 µM) and for 30 min. LPA only: treatment with LPA (5 µg/mL); P10→LPA: incubation with propofol (10 µM) and treatment with LPA (5 µg/mLl); A100 only: treatment with aminophylline (100 µM); P10→A100: incubation with propofol (10 µM) and treatment with aminophylline (100 µM) Values represent the mean peak intracellular calcium response. Results are expressed as fold-stimulation, determined by comparing relative fluorescence intensities (RFI) before stimulation and expressed as mean±SD from three separate determinations. Each determination represents the mean of at least 10 cells. (B) Levels of mean peak intracellular calcium ([Ca2+]i) generated by lysophosphatidic acid (LPA) or aminophylline treatment after incubation with propofol (30 µM) and for 30 min. LPA only: treatment with LPA (5 µg/mL); P30→LPA: incubation with propofol (30 µM) and treatment with LPA (5 µg/mL); A100 only: treatment with aminophylline (100 µM); P30→A100: incubation with propofol (30 µM) and treatment with aminophylline (100 µM) Values represent the mean peak intracellular calcium response. Results are expressed as fold-stimulation, determined by comparing relative fluorescence intensities (RFI) before stimulation and expressed as mean±SD from three separate determinations. Each determination represents the mean of at least 10 cells.*P<0.05 compared with treatment with LPA (5 µg/mL); †P<0.05 compared with treatment with aminophylline (100 µM).
Mentions: Aminophylline induced a very rapid, dose-dependent increase in [Ca2+]i (Fig. 3), and propofol (300 µM) treatment decreased the concentration of [Ca2+]i. Aminophylline (1,000 µM) rapidly increased Fluo-4 fluorescence in cells to a maximum three to four-fold higher than the control (Fig. 3). Propofol (10 µM) treatment for 30 min decreased [Ca2+]i induced by LPA (5 µg/mL) and aminophylline (100 µM) (Fig. 4A). Propofol (30 µM) showed similar activity (Fig. 4B). Following incubation with propofol (30 µM) and aminophylline (100; 1,000 µM) for 30 min, the peak level of [Ca2+]i following LPA (5 µg/mL) treatment was higher than propofol (30 µM) only (Fig. 5). Furthermore, HUVECs incubated with propofol (30 µM) and aminophylline (1,000 µM) for 30 min had peak [Ca2+]i levels higher than propofol (30 µM) and aminophylline (100 µM) (Fig. 5).

Bottom Line: The results were expressed as relative fluorescence intensity and fold stimulation.Propofol further prevented the aminophylline-induced increase of intracellular calcium at clinically relevant concentrations.However, aminophylline reversed the inhibitory effect of propofol on the elevation of intracellular calcium by lysophosphatidic acid.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesiology and Pain Medicine, Kangwon National University Medical School, Chuncheon, Korea.

ABSTRACT
This study examined whether propofol and aminophylline affect the mobilization of intracellular calcium in human umbilical vein endothelial cells. Intracellular calcium was measured using laser scanning confocal microscopy. Cultured and serum-starved cells on round coverslips were incubated with propofol or aminophylline for 30 min, and then stimulated with lysophosphatidic acid, propofol and aminophylline. The results were expressed as relative fluorescence intensity and fold stimulation. Propofol decreased the concentration of intracellular calcium, whereas aminophylline caused increased mobilization of intracellular calcium in a concentration-dependent manner. Propofol suppressed the lysophosphatidic acid-induced mobilization of intracellular calcium in a concentration-dependent manner. Propofol further prevented the aminophylline-induced increase of intracellular calcium at clinically relevant concentrations. However, aminophylline reversed the inhibitory effect of propofol on the elevation of intracellular calcium by lysophosphatidic acid. Our results suggest that propofol and aminophylline antagonize each other on the mobilization of intracellular calcium in human umbilical vein endothelial cells at clinically relevant concentrations. Serious consideration should be given to how this interaction affects mobilization of intracellular calcium when these two drugs are used together.

Show MeSH