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Anesthetic propofol overdose causes vascular hyperpermeability by reducing endothelial glycocalyx and ATP production.

Lin MC, Lin CF, Li CF, Sun DP, Wang LY, Hsing CH - Int J Mol Sci (2015)

Bottom Line: In vivo, we intraperitoneally injected ICR mice with overdosed propofol, and the results showed that a propofol overdose significantly induced systemic vascular hyperpermeability and reduced the expression of endothelial glycocalyx, syndecan-1, syndecan-4, perlecan mRNA and heparan sulfate (HS) in the vessels of multiple organs.In vitro, a propofol overdose reduced the expression of syndecan-1, syndecan-4, perlecan, glypican-1 mRNA and HS and induced significant decreases in the nicotinamide adenine dinucleotide (NAD+)/NADH ratio and ATP concentrations in human microvascular endothelial cells (HMEC-1).Oligomycin treatment also induced significant decreases in the NAD+/NADH ratio, in ATP concentrations and in syndecan-4, perlecan and glypican-1 mRNA expression in HMEC-1 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesiology, Chi Mei Medical Center, Liouying, 201, Taikang, Taikang Village, Liuying District, Tainan 736, Taiwan. mygegon@gmail.com.

ABSTRACT
Prolonged treatment with a large dose of propofol may cause diffuse cellular cytotoxicity; however, the detailed underlying mechanism remains unclear, particularly in vascular endothelial cells. Previous studies showed that a propofol overdose induces endothelial injury and vascular barrier dysfunction. Regarding the important role of endothelial glycocalyx on the maintenance of vascular barrier integrity, we therefore hypothesized that a propofol overdose-induced endothelial barrier dysfunction is caused by impaired endothelial glycocalyx. In vivo, we intraperitoneally injected ICR mice with overdosed propofol, and the results showed that a propofol overdose significantly induced systemic vascular hyperpermeability and reduced the expression of endothelial glycocalyx, syndecan-1, syndecan-4, perlecan mRNA and heparan sulfate (HS) in the vessels of multiple organs. In vitro, a propofol overdose reduced the expression of syndecan-1, syndecan-4, perlecan, glypican-1 mRNA and HS and induced significant decreases in the nicotinamide adenine dinucleotide (NAD+)/NADH ratio and ATP concentrations in human microvascular endothelial cells (HMEC-1). Oligomycin treatment also induced significant decreases in the NAD+/NADH ratio, in ATP concentrations and in syndecan-4, perlecan and glypican-1 mRNA expression in HMEC-1 cells. These results demonstrate that a propofol overdose induces a partially ATP-dependent reduction of endothelial glycocalyx expression and consequently leads to vascular hyperpermeability due to the loss of endothelial barrier functions.

No MeSH data available.


Related in: MedlinePlus

Propofol overdose effect on the relative expression of SDC-1, SDC4, PLC and GPC-1 mRNA and on HS expression in HMEC-1 cells treated with or without propofol for 24 h. (A) Cell viability was measured using an MTT assay and cytotoxicity using an LDH assay; (B) Endothelial HS expression was detected using immunocytochemical (ICC) staining followed by light microscopy (B, bottom: arrowhead). A micrograph of an apoptotic cell in propofol-treated cells is shown; (C) Endothelial HS expression was detected using Western blotting; (D) The relative expression levels of SDC-1, SDC4, PLC and GPC-1 mRNA in HMEC-1 were detected using RT-PCR. For the MTT assay and RT-PCR, the ratios to the untreated control are shown (means ± standard deviation of triplicate cultures). * p < 0.05, ** p < 0.01 compared with the control group. LDH assay data are shown as OD values (means ± SD of triplicate cultures). * p < 0.05, ** p < 0.01 compared with the untreated group.
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ijms-16-12092-f004: Propofol overdose effect on the relative expression of SDC-1, SDC4, PLC and GPC-1 mRNA and on HS expression in HMEC-1 cells treated with or without propofol for 24 h. (A) Cell viability was measured using an MTT assay and cytotoxicity using an LDH assay; (B) Endothelial HS expression was detected using immunocytochemical (ICC) staining followed by light microscopy (B, bottom: arrowhead). A micrograph of an apoptotic cell in propofol-treated cells is shown; (C) Endothelial HS expression was detected using Western blotting; (D) The relative expression levels of SDC-1, SDC4, PLC and GPC-1 mRNA in HMEC-1 were detected using RT-PCR. For the MTT assay and RT-PCR, the ratios to the untreated control are shown (means ± standard deviation of triplicate cultures). * p < 0.05, ** p < 0.01 compared with the control group. LDH assay data are shown as OD values (means ± SD of triplicate cultures). * p < 0.05, ** p < 0.01 compared with the untreated group.

Mentions: An MTT assay showed lower cell viability (Figure 4A, top) and an LDH assay showed greater cytotoxicity (Figure 4A, bottom) in HMEC-1 cells treated with 50 and with 100 μg/mL of propofol. HMEC-1 cells overdosed with propofol showed cell apoptosis (Figure 4B, bottom). To specifically clarify the effect of a propofol overdose on endothelial glycocalyx alone, we lowered the maximum dose of propofol to 20 μg/mL for HMEC-1 cells. Both the Western blot (Figure 4C) and the light microscope images of immunocytochemical (ICC) staining (Figure 4B) showed that endothelial HS was significantly lower in HMEC-1 cells treated with 20 μg/mL of propofol. The full film of another Western blot was shown (Supplementary Data, Figure S2). In addition, RT-PCR showed that the relative expression of syndecan-1, syndecan-4 and glypican-1 mRNA was significantly (p < 0.05) lower than in the control cells. The relative expression of perlecan mRNA was even markedly (p < 0.05) reduced by propofol at doses above 10 μg/mL (Figure 4D).


Anesthetic propofol overdose causes vascular hyperpermeability by reducing endothelial glycocalyx and ATP production.

Lin MC, Lin CF, Li CF, Sun DP, Wang LY, Hsing CH - Int J Mol Sci (2015)

Propofol overdose effect on the relative expression of SDC-1, SDC4, PLC and GPC-1 mRNA and on HS expression in HMEC-1 cells treated with or without propofol for 24 h. (A) Cell viability was measured using an MTT assay and cytotoxicity using an LDH assay; (B) Endothelial HS expression was detected using immunocytochemical (ICC) staining followed by light microscopy (B, bottom: arrowhead). A micrograph of an apoptotic cell in propofol-treated cells is shown; (C) Endothelial HS expression was detected using Western blotting; (D) The relative expression levels of SDC-1, SDC4, PLC and GPC-1 mRNA in HMEC-1 were detected using RT-PCR. For the MTT assay and RT-PCR, the ratios to the untreated control are shown (means ± standard deviation of triplicate cultures). * p < 0.05, ** p < 0.01 compared with the control group. LDH assay data are shown as OD values (means ± SD of triplicate cultures). * p < 0.05, ** p < 0.01 compared with the untreated group.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4490431&req=5

ijms-16-12092-f004: Propofol overdose effect on the relative expression of SDC-1, SDC4, PLC and GPC-1 mRNA and on HS expression in HMEC-1 cells treated with or without propofol for 24 h. (A) Cell viability was measured using an MTT assay and cytotoxicity using an LDH assay; (B) Endothelial HS expression was detected using immunocytochemical (ICC) staining followed by light microscopy (B, bottom: arrowhead). A micrograph of an apoptotic cell in propofol-treated cells is shown; (C) Endothelial HS expression was detected using Western blotting; (D) The relative expression levels of SDC-1, SDC4, PLC and GPC-1 mRNA in HMEC-1 were detected using RT-PCR. For the MTT assay and RT-PCR, the ratios to the untreated control are shown (means ± standard deviation of triplicate cultures). * p < 0.05, ** p < 0.01 compared with the control group. LDH assay data are shown as OD values (means ± SD of triplicate cultures). * p < 0.05, ** p < 0.01 compared with the untreated group.
Mentions: An MTT assay showed lower cell viability (Figure 4A, top) and an LDH assay showed greater cytotoxicity (Figure 4A, bottom) in HMEC-1 cells treated with 50 and with 100 μg/mL of propofol. HMEC-1 cells overdosed with propofol showed cell apoptosis (Figure 4B, bottom). To specifically clarify the effect of a propofol overdose on endothelial glycocalyx alone, we lowered the maximum dose of propofol to 20 μg/mL for HMEC-1 cells. Both the Western blot (Figure 4C) and the light microscope images of immunocytochemical (ICC) staining (Figure 4B) showed that endothelial HS was significantly lower in HMEC-1 cells treated with 20 μg/mL of propofol. The full film of another Western blot was shown (Supplementary Data, Figure S2). In addition, RT-PCR showed that the relative expression of syndecan-1, syndecan-4 and glypican-1 mRNA was significantly (p < 0.05) lower than in the control cells. The relative expression of perlecan mRNA was even markedly (p < 0.05) reduced by propofol at doses above 10 μg/mL (Figure 4D).

Bottom Line: In vivo, we intraperitoneally injected ICR mice with overdosed propofol, and the results showed that a propofol overdose significantly induced systemic vascular hyperpermeability and reduced the expression of endothelial glycocalyx, syndecan-1, syndecan-4, perlecan mRNA and heparan sulfate (HS) in the vessels of multiple organs.In vitro, a propofol overdose reduced the expression of syndecan-1, syndecan-4, perlecan, glypican-1 mRNA and HS and induced significant decreases in the nicotinamide adenine dinucleotide (NAD+)/NADH ratio and ATP concentrations in human microvascular endothelial cells (HMEC-1).Oligomycin treatment also induced significant decreases in the NAD+/NADH ratio, in ATP concentrations and in syndecan-4, perlecan and glypican-1 mRNA expression in HMEC-1 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesiology, Chi Mei Medical Center, Liouying, 201, Taikang, Taikang Village, Liuying District, Tainan 736, Taiwan. mygegon@gmail.com.

ABSTRACT
Prolonged treatment with a large dose of propofol may cause diffuse cellular cytotoxicity; however, the detailed underlying mechanism remains unclear, particularly in vascular endothelial cells. Previous studies showed that a propofol overdose induces endothelial injury and vascular barrier dysfunction. Regarding the important role of endothelial glycocalyx on the maintenance of vascular barrier integrity, we therefore hypothesized that a propofol overdose-induced endothelial barrier dysfunction is caused by impaired endothelial glycocalyx. In vivo, we intraperitoneally injected ICR mice with overdosed propofol, and the results showed that a propofol overdose significantly induced systemic vascular hyperpermeability and reduced the expression of endothelial glycocalyx, syndecan-1, syndecan-4, perlecan mRNA and heparan sulfate (HS) in the vessels of multiple organs. In vitro, a propofol overdose reduced the expression of syndecan-1, syndecan-4, perlecan, glypican-1 mRNA and HS and induced significant decreases in the nicotinamide adenine dinucleotide (NAD+)/NADH ratio and ATP concentrations in human microvascular endothelial cells (HMEC-1). Oligomycin treatment also induced significant decreases in the NAD+/NADH ratio, in ATP concentrations and in syndecan-4, perlecan and glypican-1 mRNA expression in HMEC-1 cells. These results demonstrate that a propofol overdose induces a partially ATP-dependent reduction of endothelial glycocalyx expression and consequently leads to vascular hyperpermeability due to the loss of endothelial barrier functions.

No MeSH data available.


Related in: MedlinePlus