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Various Molecular Species of Diacylglycerol Hydroperoxide Activate Human Neutrophils via PKC Activation.

Kambayashi Y, Takekoshi S, Tanino Y, Watanabe K, Nakano M, Hitomi Y, Takigawa T, Ogino K, Yamamoto Y - J Clin Biochem Nutr (2007)

Bottom Line: We have proposed that diacylglycerol hydroperoxide-induced unregulated signal transduction causes oxidative stress-related diseases.Moreover, the time course of p47(phox) phosphorylation was comparable to that of superoxide production.These results suggest that PLG-OOH activated intracellular protein kinase C.

View Article: PubMed Central - PubMed

Affiliation: Department of Environmental and Preventive Medicine, Graduate School of Medical Science, Kanazawa University, 13-1 Takara-machi, Kanazawa 920-8640, Japan.

ABSTRACT
We have proposed that diacylglycerol hydroperoxide-induced unregulated signal transduction causes oxidative stress-related diseases. In this study, we investigated which molecular species of diacylglycerol hydroperoxide activated human peripheral neutrophils. All diacylglycerol hydroperoxides, diacylglycerol hydroxides, and diacyglycerols tested in the present study induced superoxide production by neutrophils. The ability to activate neutrophils among molecular species containing the same fatty acid composition was as follows; diacylglycerol hydroperoxide>diacylglycerol hydroxide>/=diacylglycerol. The diacylglycerol hydroperoxide composed of linoleate was a stronger activator for neutrophils than that composed of arachidonate. 1-Palmitoyl-2-linoleoylglycerol hydroperoxide (PLG-OOH) was the strongest stimulator for neutrophils. We reconfirmed that PLG-OOH activated protein kinase C (PKC) in neutrophils. PLG-OOH induced the phosphorylation of p47(phox), a substrate of PKC and a cytosolic component of NADPH oxidase, in neutrophils, as did N-formyl-methionyl-leucyl-phenylalanine or 4beta-phorbol-12beta-myristate-13alpha-acetate. Moreover, the time course of p47(phox) phosphorylation was comparable to that of superoxide production. These results suggest that PLG-OOH activated intracellular protein kinase C. PLG-OOH, produced via an uncontrolled process, can act as a biological second messenger to cause inflammatory disease from oxidative stress.

No MeSH data available.


Related in: MedlinePlus

Phosphorylation of p47phox in neutrophils (3 × 106 cells/ml) activated by various stimulators. Lane 1; Methanol (0.5% of solution) was used in the control experiment (2 min). Lane 2; Three nanomolar 4β-phorbol-12β-myristate-13α-acetate (PMA) was used, and stimulation was stopped at 5 min. Lane 3; One micromolar N-formyl-methionyl-1-leucyl-phenylalnine (fMLP) was used, and stimulation was stopped at 2 min. Lane 4; Eight micromolar 1-palmitoyl-2-linoleoylglycerol hydroperoxide (PLG-OOH) was used, and stimulation was stopped at 2 min.
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Figure 4: Phosphorylation of p47phox in neutrophils (3 × 106 cells/ml) activated by various stimulators. Lane 1; Methanol (0.5% of solution) was used in the control experiment (2 min). Lane 2; Three nanomolar 4β-phorbol-12β-myristate-13α-acetate (PMA) was used, and stimulation was stopped at 5 min. Lane 3; One micromolar N-formyl-methionyl-1-leucyl-phenylalnine (fMLP) was used, and stimulation was stopped at 2 min. Lane 4; Eight micromolar 1-palmitoyl-2-linoleoylglycerol hydroperoxide (PLG-OOH) was used, and stimulation was stopped at 2 min.

Mentions: The activation of PKC in neutrophils by DAG-OOH was reconfirmed. p47phox is a cytosolic factor of NADPH oxidase, and known as a substrate of PKC [21]. When only methanol (0.5% to solution) was added to the neutrophil suspension, p47phox was not phosphorylated after 2 min at 37°C (Fig. 4). p47phox was phosphorylated in 8 µM PLG-OOH-stimulated neutrophils, as in PMA (3 nM)-stimulated or fMLP (1 µM)-stimulated neutrophils (Fig. 4).


Various Molecular Species of Diacylglycerol Hydroperoxide Activate Human Neutrophils via PKC Activation.

Kambayashi Y, Takekoshi S, Tanino Y, Watanabe K, Nakano M, Hitomi Y, Takigawa T, Ogino K, Yamamoto Y - J Clin Biochem Nutr (2007)

Phosphorylation of p47phox in neutrophils (3 × 106 cells/ml) activated by various stimulators. Lane 1; Methanol (0.5% of solution) was used in the control experiment (2 min). Lane 2; Three nanomolar 4β-phorbol-12β-myristate-13α-acetate (PMA) was used, and stimulation was stopped at 5 min. Lane 3; One micromolar N-formyl-methionyl-1-leucyl-phenylalnine (fMLP) was used, and stimulation was stopped at 2 min. Lane 4; Eight micromolar 1-palmitoyl-2-linoleoylglycerol hydroperoxide (PLG-OOH) was used, and stimulation was stopped at 2 min.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2274990&req=5

Figure 4: Phosphorylation of p47phox in neutrophils (3 × 106 cells/ml) activated by various stimulators. Lane 1; Methanol (0.5% of solution) was used in the control experiment (2 min). Lane 2; Three nanomolar 4β-phorbol-12β-myristate-13α-acetate (PMA) was used, and stimulation was stopped at 5 min. Lane 3; One micromolar N-formyl-methionyl-1-leucyl-phenylalnine (fMLP) was used, and stimulation was stopped at 2 min. Lane 4; Eight micromolar 1-palmitoyl-2-linoleoylglycerol hydroperoxide (PLG-OOH) was used, and stimulation was stopped at 2 min.
Mentions: The activation of PKC in neutrophils by DAG-OOH was reconfirmed. p47phox is a cytosolic factor of NADPH oxidase, and known as a substrate of PKC [21]. When only methanol (0.5% to solution) was added to the neutrophil suspension, p47phox was not phosphorylated after 2 min at 37°C (Fig. 4). p47phox was phosphorylated in 8 µM PLG-OOH-stimulated neutrophils, as in PMA (3 nM)-stimulated or fMLP (1 µM)-stimulated neutrophils (Fig. 4).

Bottom Line: We have proposed that diacylglycerol hydroperoxide-induced unregulated signal transduction causes oxidative stress-related diseases.Moreover, the time course of p47(phox) phosphorylation was comparable to that of superoxide production.These results suggest that PLG-OOH activated intracellular protein kinase C.

View Article: PubMed Central - PubMed

Affiliation: Department of Environmental and Preventive Medicine, Graduate School of Medical Science, Kanazawa University, 13-1 Takara-machi, Kanazawa 920-8640, Japan.

ABSTRACT
We have proposed that diacylglycerol hydroperoxide-induced unregulated signal transduction causes oxidative stress-related diseases. In this study, we investigated which molecular species of diacylglycerol hydroperoxide activated human peripheral neutrophils. All diacylglycerol hydroperoxides, diacylglycerol hydroxides, and diacyglycerols tested in the present study induced superoxide production by neutrophils. The ability to activate neutrophils among molecular species containing the same fatty acid composition was as follows; diacylglycerol hydroperoxide>diacylglycerol hydroxide>/=diacylglycerol. The diacylglycerol hydroperoxide composed of linoleate was a stronger activator for neutrophils than that composed of arachidonate. 1-Palmitoyl-2-linoleoylglycerol hydroperoxide (PLG-OOH) was the strongest stimulator for neutrophils. We reconfirmed that PLG-OOH activated protein kinase C (PKC) in neutrophils. PLG-OOH induced the phosphorylation of p47(phox), a substrate of PKC and a cytosolic component of NADPH oxidase, in neutrophils, as did N-formyl-methionyl-leucyl-phenylalanine or 4beta-phorbol-12beta-myristate-13alpha-acetate. Moreover, the time course of p47(phox) phosphorylation was comparable to that of superoxide production. These results suggest that PLG-OOH activated intracellular protein kinase C. PLG-OOH, produced via an uncontrolled process, can act as a biological second messenger to cause inflammatory disease from oxidative stress.

No MeSH data available.


Related in: MedlinePlus