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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 cell apoptosis 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. PI and PI/Annexin V-FITC staining followed by flow cytometric analysis determined the apoptosis levels. The percentages of apoptotic cells (sub-G1 and Annexin V+/PI-) are shown as the means ± SD of three individual experiments. **P < 0.01 and ***P < 0.001 compared with untreated at 0 h. #P < 0.05 and ##P < 0.01 compared with untreated at each time points. †P < 0.05 and ††P < 0.01 compared with propofol. ns, not significant compared with propofol plus LY294002.
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pone.0129693.g005: PI3-kinase-dependent propofol treatment abolishes cell apoptosis 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. PI and PI/Annexin V-FITC staining followed by flow cytometric analysis determined the apoptosis levels. The percentages of apoptotic cells (sub-G1 and Annexin V+/PI-) are shown as the means ± SD of three individual experiments. **P < 0.01 and ***P < 0.001 compared with untreated at 0 h. #P < 0.05 and ##P < 0.01 compared with untreated at each time points. †P < 0.05 and ††P < 0.01 compared with propofol. ns, not significant compared with propofol plus LY294002.

Mentions: To verify the effects of propofol on PI3-kinase activity, the generation of PIP3 confirmed the inhibitory effect of propofol on PI3-kinase inactivation in primary neutrophils (Fig 4A) and granulocyte-differentiated HL60 cells (Fig 4B). These results indicate that propofol may retard PI3-kinase down-regulation in neutrophils undergoing apoptosis. We next hypothesized that a pro-survival role of propofol prevents neutrophil apoptosis by sustaining PI3-kinase activity. Nuclear PI staining followed by flow cytometric analysis showed that the primary neutrophils and granuloctyte-differentiated HL60 cells underwent apoptosis (as shown in the sub-G1 phase), whereas propofol significantly prevented the apoptotic effects in a PI3-kinase-mediated manner (Fig 5A and 5B, top). PI/Annexin V-FITC staining confirmed the effects of propofol on preventing early apoptosis (Fig 5A and 5B, bottom). These results demonstrate the important role of propofol in maintaining PI3-kinase activity.


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 cell apoptosis 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. PI and PI/Annexin V-FITC staining followed by flow cytometric analysis determined the apoptosis levels. The percentages of apoptotic cells (sub-G1 and Annexin V+/PI-) are shown as the means ± SD of three individual experiments. **P < 0.01 and ***P < 0.001 compared with untreated at 0 h. #P < 0.05 and ##P < 0.01 compared with untreated at each time points. †P < 0.05 and ††P < 0.01 compared with propofol. ns, not significant compared with propofol plus LY294002.
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pone.0129693.g005: PI3-kinase-dependent propofol treatment abolishes cell apoptosis 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. PI and PI/Annexin V-FITC staining followed by flow cytometric analysis determined the apoptosis levels. The percentages of apoptotic cells (sub-G1 and Annexin V+/PI-) are shown as the means ± SD of three individual experiments. **P < 0.01 and ***P < 0.001 compared with untreated at 0 h. #P < 0.05 and ##P < 0.01 compared with untreated at each time points. †P < 0.05 and ††P < 0.01 compared with propofol. ns, not significant compared with propofol plus LY294002.
Mentions: To verify the effects of propofol on PI3-kinase activity, the generation of PIP3 confirmed the inhibitory effect of propofol on PI3-kinase inactivation in primary neutrophils (Fig 4A) and granulocyte-differentiated HL60 cells (Fig 4B). These results indicate that propofol may retard PI3-kinase down-regulation in neutrophils undergoing apoptosis. We next hypothesized that a pro-survival role of propofol prevents neutrophil apoptosis by sustaining PI3-kinase activity. Nuclear PI staining followed by flow cytometric analysis showed that the primary neutrophils and granuloctyte-differentiated HL60 cells underwent apoptosis (as shown in the sub-G1 phase), whereas propofol significantly prevented the apoptotic effects in a PI3-kinase-mediated manner (Fig 5A and 5B, top). PI/Annexin V-FITC staining confirmed the effects of propofol on preventing early apoptosis (Fig 5A and 5B, bottom). These results demonstrate the important role of propofol in maintaining PI3-kinase activity.

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