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Propofol Treatment Inhibits Constitutive Apoptosis in Human Primary Neutrophils and Granulocyte-Differentiated Human HL60 Cells.

Hsing CH, Chen CL, Lin WC, Lin CF - PLoS ONE (2015)

Bottom Line: The inactivation of phosphatidylinositol 3-kinase (PI3-kinase)/AKT and the activation of GSK-3β results in myeloid cell leukemia 1 (Mcl-1) down-regulation, the loss of the mitochondrial transmembrane potential, and caspase-3 activation in these cells, which is accompanied by apoptosis.Notably, propofol treatment attenuates these effects in a PI3-kinase-regulated manner.We found that propofol initiates PI3-kinase/AKT-mediated GSK-3β inactivation and Mcl-1 stabilization, rescuing the constitutive apoptosis in primary neutrophils and granulocyte-differentiated acute promyelocytic leukemia HL60 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesiology, Chi Mei Medical Center, Tainan, Taiwan; Department of Anesthesiology, College of Medicine, Taipei Medical University, Taipei, Taiwan.

ABSTRACT
Apoptosis regulation is essential for neutrophil homeostasis. We previously demonstrated that a process involving glycogen synthase kinase (GSK)-3β determines neutrophil apoptosis. As for this apoptotic process, an overdose of propofol (2,6-Diisopropylphenol; 25 μg/ml or 140 μM) also causes GSK-3β-mediated macrophage apoptosis; however, the early deactivation of GSK-3β with low-dose propofol has been shown. Therefore, we hypothesize that low-dose propofol may induce neutrophil survival via GSK-3β inactivation. Following in vitro culture, the therapeutic concentration of propofol (10 μg/ml or 56 μM) treatment decreased constitutive apoptosis in isolated human primary neutrophils and in granulocyte-differentiated HL60 cells after all-trans retinoic acid (1 μM) treatment. The inactivation of phosphatidylinositol 3-kinase (PI3-kinase)/AKT and the activation of GSK-3β results in myeloid cell leukemia 1 (Mcl-1) down-regulation, the loss of the mitochondrial transmembrane potential, and caspase-3 activation in these cells, which is accompanied by apoptosis. Notably, propofol treatment attenuates these effects in a PI3-kinase-regulated manner. We found that propofol initiates PI3-kinase/AKT-mediated GSK-3β inactivation and Mcl-1 stabilization, rescuing the constitutive apoptosis in primary neutrophils and granulocyte-differentiated acute promyelocytic leukemia HL60 cells.

No MeSH data available.


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PI3-kinase-dependent propofol treatment abolishes AKT inactivation and GSK-3β activation in primary neutrophils and ATRA-differentiated granulocytic HL60 cells.(A) Isolated human primary neutrophils and (B) ATRA (1 μM)-differentiated granulocytic HL60 cells were treated with propofol with or without LY294002 treatment for the indicated time points. Western blot analysis showing the expression of phospho-AKT (Ser473), AKT, phospho-GSK-3β (Ser9), phospho-GSK-3β (Tyr216), and GSK3-β; β-actin was used as the loading control. One representative dataset obtained from three individual experiments is shown. The changes in the ratio of the phosphorylated protein and its total proteins are also shown. The data are shown as the means ± SD of three individual experiments. *P < 0.05 compared with untreated at 0 h. #P < 0.05 compared with untreated at 24 h or day 6. †P < 0.05 compared with propofol.
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pone.0129693.g003: PI3-kinase-dependent propofol treatment abolishes AKT inactivation and GSK-3β activation in primary neutrophils and ATRA-differentiated granulocytic HL60 cells.(A) Isolated human primary neutrophils and (B) ATRA (1 μM)-differentiated granulocytic HL60 cells were treated with propofol with or without LY294002 treatment for the indicated time points. Western blot analysis showing the expression of phospho-AKT (Ser473), AKT, phospho-GSK-3β (Ser9), phospho-GSK-3β (Tyr216), and GSK3-β; β-actin was used as the loading control. One representative dataset obtained from three individual experiments is shown. The changes in the ratio of the phosphorylated protein and its total proteins are also shown. The data are shown as the means ± SD of three individual experiments. *P < 0.05 compared with untreated at 0 h. #P < 0.05 compared with untreated at 24 h or day 6. †P < 0.05 compared with propofol.

Mentions: Because the activation of GSK-3β is required for neutrophils undergoing apoptosis [11], we next examined whether propofol inhibits GSK-3β activation by altering PI3-kinase/AKT signaling [7,8]. Western blot analysis revealed AKT dephosphorylation (Ser473), followed by GSK-3β dephosphorylation (Ser9, an inactive form of GSK-3β) and phosphorylation (Tyr216, an active form of GSK-3β) in 24 h post-culture while propofol significantly reduced such the effects in primary neutrophils (Fig 3A) and granulocyte-differentiated HL60 cells (Fig 3B). Importantly, PI3-kinase inhibitor LY294002 significantly reversed these effects induced by propofol. These findings demonstrate that propofol induces PI3-kinase/AKT-mediated GSK-3β inactivation.


Propofol Treatment Inhibits Constitutive Apoptosis in Human Primary Neutrophils and Granulocyte-Differentiated Human HL60 Cells.

Hsing CH, Chen CL, Lin WC, Lin CF - PLoS ONE (2015)

PI3-kinase-dependent propofol treatment abolishes AKT inactivation and GSK-3β activation in primary neutrophils and ATRA-differentiated granulocytic HL60 cells.(A) Isolated human primary neutrophils and (B) ATRA (1 μM)-differentiated granulocytic HL60 cells were treated with propofol with or without LY294002 treatment for the indicated time points. Western blot analysis showing the expression of phospho-AKT (Ser473), AKT, phospho-GSK-3β (Ser9), phospho-GSK-3β (Tyr216), and GSK3-β; β-actin was used as the loading control. One representative dataset obtained from three individual experiments is shown. The changes in the ratio of the phosphorylated protein and its total proteins are also shown. The data are shown as the means ± SD of three individual experiments. *P < 0.05 compared with untreated at 0 h. #P < 0.05 compared with untreated at 24 h or day 6. †P < 0.05 compared with propofol.
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pone.0129693.g003: PI3-kinase-dependent propofol treatment abolishes AKT inactivation and GSK-3β activation in primary neutrophils and ATRA-differentiated granulocytic HL60 cells.(A) Isolated human primary neutrophils and (B) ATRA (1 μM)-differentiated granulocytic HL60 cells were treated with propofol with or without LY294002 treatment for the indicated time points. Western blot analysis showing the expression of phospho-AKT (Ser473), AKT, phospho-GSK-3β (Ser9), phospho-GSK-3β (Tyr216), and GSK3-β; β-actin was used as the loading control. One representative dataset obtained from three individual experiments is shown. The changes in the ratio of the phosphorylated protein and its total proteins are also shown. The data are shown as the means ± SD of three individual experiments. *P < 0.05 compared with untreated at 0 h. #P < 0.05 compared with untreated at 24 h or day 6. †P < 0.05 compared with propofol.
Mentions: Because the activation of GSK-3β is required for neutrophils undergoing apoptosis [11], we next examined whether propofol inhibits GSK-3β activation by altering PI3-kinase/AKT signaling [7,8]. Western blot analysis revealed AKT dephosphorylation (Ser473), followed by GSK-3β dephosphorylation (Ser9, an inactive form of GSK-3β) and phosphorylation (Tyr216, an active form of GSK-3β) in 24 h post-culture while propofol significantly reduced such the effects in primary neutrophils (Fig 3A) and granulocyte-differentiated HL60 cells (Fig 3B). Importantly, PI3-kinase inhibitor LY294002 significantly reversed these effects induced by propofol. These findings demonstrate that propofol induces PI3-kinase/AKT-mediated GSK-3β inactivation.

Bottom Line: The inactivation of phosphatidylinositol 3-kinase (PI3-kinase)/AKT and the activation of GSK-3β results in myeloid cell leukemia 1 (Mcl-1) down-regulation, the loss of the mitochondrial transmembrane potential, and caspase-3 activation in these cells, which is accompanied by apoptosis.Notably, propofol treatment attenuates these effects in a PI3-kinase-regulated manner.We found that propofol initiates PI3-kinase/AKT-mediated GSK-3β inactivation and Mcl-1 stabilization, rescuing the constitutive apoptosis in primary neutrophils and granulocyte-differentiated acute promyelocytic leukemia HL60 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesiology, Chi Mei Medical Center, Tainan, Taiwan; Department of Anesthesiology, College of Medicine, Taipei Medical University, Taipei, Taiwan.

ABSTRACT
Apoptosis regulation is essential for neutrophil homeostasis. We previously demonstrated that a process involving glycogen synthase kinase (GSK)-3β determines neutrophil apoptosis. As for this apoptotic process, an overdose of propofol (2,6-Diisopropylphenol; 25 μg/ml or 140 μM) also causes GSK-3β-mediated macrophage apoptosis; however, the early deactivation of GSK-3β with low-dose propofol has been shown. Therefore, we hypothesize that low-dose propofol may induce neutrophil survival via GSK-3β inactivation. Following in vitro culture, the therapeutic concentration of propofol (10 μg/ml or 56 μM) treatment decreased constitutive apoptosis in isolated human primary neutrophils and in granulocyte-differentiated HL60 cells after all-trans retinoic acid (1 μM) treatment. The inactivation of phosphatidylinositol 3-kinase (PI3-kinase)/AKT and the activation of GSK-3β results in myeloid cell leukemia 1 (Mcl-1) down-regulation, the loss of the mitochondrial transmembrane potential, and caspase-3 activation in these cells, which is accompanied by apoptosis. Notably, propofol treatment attenuates these effects in a PI3-kinase-regulated manner. We found that propofol initiates PI3-kinase/AKT-mediated GSK-3β inactivation and Mcl-1 stabilization, rescuing the constitutive apoptosis in primary neutrophils and granulocyte-differentiated acute promyelocytic leukemia HL60 cells.

No MeSH data available.


Related in: MedlinePlus