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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 Mcl-1 down-regulation 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 Mcl-1; β-actin was used as the loading control. One representative dataset obtained from three individual experiments is shown. The changes in the ratio of Mcl-1 and β-actin 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. ns, not significant compared with untreated at 24 h or day 6.
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pone.0129693.g006: PI3-kinase-dependent propofol treatment abolishes Mcl-1 down-regulation 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 Mcl-1; β-actin was used as the loading control. One representative dataset obtained from three individual experiments is shown. The changes in the ratio of Mcl-1 and β-actin 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. ns, not significant compared with untreated at 24 h or day 6.

Mentions: GSK-3β activation causes Mcl-1 downregulation, facilitating neutrophil apoptosis, as demonstrated in our previous studies. We next investigated the effects of propofol on the expression of anti-apoptotic Mcl-1. Western blot analysis showed that propofol treatment effectively reversed the destabilization of Mcl-1 in primary neutrophils (Fig 6A) and granulocyte-differentiated HL60 cells (Fig 6B). Furthermore, pharmacologically inhibiting PI3-kinase reversed the effects of propofol. These results indicate that the PI3-kinase/AKT/GSK-3β-regulated Mcl-1 determines the constitutive apoptosis of neutrophils.


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 Mcl-1 down-regulation 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 Mcl-1; β-actin was used as the loading control. One representative dataset obtained from three individual experiments is shown. The changes in the ratio of Mcl-1 and β-actin 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. ns, not significant compared with untreated at 24 h or day 6.
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pone.0129693.g006: PI3-kinase-dependent propofol treatment abolishes Mcl-1 down-regulation 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 Mcl-1; β-actin was used as the loading control. One representative dataset obtained from three individual experiments is shown. The changes in the ratio of Mcl-1 and β-actin 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. ns, not significant compared with untreated at 24 h or day 6.
Mentions: GSK-3β activation causes Mcl-1 downregulation, facilitating neutrophil apoptosis, as demonstrated in our previous studies. We next investigated the effects of propofol on the expression of anti-apoptotic Mcl-1. Western blot analysis showed that propofol treatment effectively reversed the destabilization of Mcl-1 in primary neutrophils (Fig 6A) and granulocyte-differentiated HL60 cells (Fig 6B). Furthermore, pharmacologically inhibiting PI3-kinase reversed the effects of propofol. These results indicate that the PI3-kinase/AKT/GSK-3β-regulated Mcl-1 determines the constitutive apoptosis of neutrophils.

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