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Dibenzoylmethane exerts metabolic activity through regulation of AMP-activated protein kinase (AMPK)-mediated glucose uptake and adipogenesis pathways.

Kim N, Kim HM, Lee ES, Lee JO, Lee HJ, Lee SK, Moon JW, Kim JH, Kim JK, Kim SJ, Park SH, Chung CH, Kim HS - PLoS ONE (2015)

Bottom Line: Dibenzoylmethane (DBM) has been shown to exert a variety of beneficial effects on human health.In pre-adipocyte cells, DBM decreased the activity of acetyl-CoA carboxylase (ACC), the rate-limiting enzyme of fatty acid synthesis.These results showed that the beneficial metabolic effects of DBM might be due to regulation of glucose uptake via AMPK in skeletal muscle and inhibition of adipogenesis in pre-adipocytes.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Korea University College of Medicine, Seoul 136-701, Korea.

ABSTRACT
Dibenzoylmethane (DBM) has been shown to exert a variety of beneficial effects on human health. However, the mechanism of action is poorly understood. In this study, DBM increased phosphorylation of AMP-activated protein kinase (AMPK) and stimulated glucose uptake in a skeletal muscle cell line. Both knockdown of AMPK with siRNA and inhibition with AMPK inhibitor blocked DBM-induced glucose uptake. DBM increased the concentration of intracellular calcium and glucose uptake due to DBM was abolished by STO-609 (a calcium/calmodulin-dependent protein kinase inhibitor). DBM stimulated phosphorylation of p38 mitogen-activated protein kinase (p38 MAPK), which was blocked by pretreatment with compound C, an AMPK inhibitor. The expression of glucose transporter type 4 (GLUT4) was increased by DBM. The translocation of GLUT4 to the plasma membrane was also increased by DBM in AMPK dependently. In addition, DBM suppressed weight gain and prevented fat accumulation in the liver and abdomen in mice fed a high-fat diet. In pre-adipocyte cells, DBM decreased the activity of acetyl-CoA carboxylase (ACC), the rate-limiting enzyme of fatty acid synthesis. Expression of the adipogenic gene, fatty acid synthase (FAS), was suppressed by DBM in an AMPK-dependent manner. These results showed that the beneficial metabolic effects of DBM might be due to regulation of glucose uptake via AMPK in skeletal muscle and inhibition of adipogenesis in pre-adipocytes.

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(A) C2C12 muscle cells pre-treated with Fluo-3, AM for 30 min were then treated with DBM (30 μM).The green fluorescent signal was detected using confocal microscopy. (B) C2C12 cells pre-treated with STO-609, a CaMKK inhibitor, for 30 min and then treated with 30 μM DBM for 1 h. The cells were then lysed with lysis buffer, and the phosphorylation of AMPKα2 was assessed by western blot using phosphorylation-specific antibodies. The level of total AMPKα2 was also assessed as a control for protein loading. * p < 0.05, as compared with basal condition. (C) Myoblast L6 cells were differentiated for 7 days and then pre-treated with STO-609 (1 μM) and then DBM (30 μM) for 1 h. Glucose uptake was then assayed for 2-DG uptake as described in the Methods. *p < 0.05, compared with control. **p < 0.05, compared with DBM-treated cells.
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pone.0120104.g003: (A) C2C12 muscle cells pre-treated with Fluo-3, AM for 30 min were then treated with DBM (30 μM).The green fluorescent signal was detected using confocal microscopy. (B) C2C12 cells pre-treated with STO-609, a CaMKK inhibitor, for 30 min and then treated with 30 μM DBM for 1 h. The cells were then lysed with lysis buffer, and the phosphorylation of AMPKα2 was assessed by western blot using phosphorylation-specific antibodies. The level of total AMPKα2 was also assessed as a control for protein loading. * p < 0.05, as compared with basal condition. (C) Myoblast L6 cells were differentiated for 7 days and then pre-treated with STO-609 (1 μM) and then DBM (30 μM) for 1 h. Glucose uptake was then assayed for 2-DG uptake as described in the Methods. *p < 0.05, compared with control. **p < 0.05, compared with DBM-treated cells.

Mentions: To further characterize the upstream components of the AMPK pathway, intracellular calcium levels were measured. To measure intracellular calcium, the fluorescence of the calcium-binding dye, fluo-3 AM, was measured. DBM increased fluorescence intensity in L6 cells (Fig. 3A), indicating that intracellular calcium concentration was increased by treatment with DBM. These results suggested CaMKK as a candidate for upstream signaling of AMPK, since CaMKK is activated by Ca2+/calmodulin binding. To test this hypothesis, C2C12 cells were pre-treated with STO-609, a CaMKK inhibitor, prior to the addition of DBM. STO-609 blocked DBM-induced AMPK phosphorylation (Fig. 3B). Moreover, pre-treatment with STO-609 blocked DBM-induced 2-DG uptake (Fig. 3C), confirming that DBM increased glucose uptake via a calcium-mediated CaMKK/AMPK pathway.


Dibenzoylmethane exerts metabolic activity through regulation of AMP-activated protein kinase (AMPK)-mediated glucose uptake and adipogenesis pathways.

Kim N, Kim HM, Lee ES, Lee JO, Lee HJ, Lee SK, Moon JW, Kim JH, Kim JK, Kim SJ, Park SH, Chung CH, Kim HS - PLoS ONE (2015)

(A) C2C12 muscle cells pre-treated with Fluo-3, AM for 30 min were then treated with DBM (30 μM).The green fluorescent signal was detected using confocal microscopy. (B) C2C12 cells pre-treated with STO-609, a CaMKK inhibitor, for 30 min and then treated with 30 μM DBM for 1 h. The cells were then lysed with lysis buffer, and the phosphorylation of AMPKα2 was assessed by western blot using phosphorylation-specific antibodies. The level of total AMPKα2 was also assessed as a control for protein loading. * p < 0.05, as compared with basal condition. (C) Myoblast L6 cells were differentiated for 7 days and then pre-treated with STO-609 (1 μM) and then DBM (30 μM) for 1 h. Glucose uptake was then assayed for 2-DG uptake as described in the Methods. *p < 0.05, compared with control. **p < 0.05, compared with DBM-treated cells.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4355612&req=5

pone.0120104.g003: (A) C2C12 muscle cells pre-treated with Fluo-3, AM for 30 min were then treated with DBM (30 μM).The green fluorescent signal was detected using confocal microscopy. (B) C2C12 cells pre-treated with STO-609, a CaMKK inhibitor, for 30 min and then treated with 30 μM DBM for 1 h. The cells were then lysed with lysis buffer, and the phosphorylation of AMPKα2 was assessed by western blot using phosphorylation-specific antibodies. The level of total AMPKα2 was also assessed as a control for protein loading. * p < 0.05, as compared with basal condition. (C) Myoblast L6 cells were differentiated for 7 days and then pre-treated with STO-609 (1 μM) and then DBM (30 μM) for 1 h. Glucose uptake was then assayed for 2-DG uptake as described in the Methods. *p < 0.05, compared with control. **p < 0.05, compared with DBM-treated cells.
Mentions: To further characterize the upstream components of the AMPK pathway, intracellular calcium levels were measured. To measure intracellular calcium, the fluorescence of the calcium-binding dye, fluo-3 AM, was measured. DBM increased fluorescence intensity in L6 cells (Fig. 3A), indicating that intracellular calcium concentration was increased by treatment with DBM. These results suggested CaMKK as a candidate for upstream signaling of AMPK, since CaMKK is activated by Ca2+/calmodulin binding. To test this hypothesis, C2C12 cells were pre-treated with STO-609, a CaMKK inhibitor, prior to the addition of DBM. STO-609 blocked DBM-induced AMPK phosphorylation (Fig. 3B). Moreover, pre-treatment with STO-609 blocked DBM-induced 2-DG uptake (Fig. 3C), confirming that DBM increased glucose uptake via a calcium-mediated CaMKK/AMPK pathway.

Bottom Line: Dibenzoylmethane (DBM) has been shown to exert a variety of beneficial effects on human health.In pre-adipocyte cells, DBM decreased the activity of acetyl-CoA carboxylase (ACC), the rate-limiting enzyme of fatty acid synthesis.These results showed that the beneficial metabolic effects of DBM might be due to regulation of glucose uptake via AMPK in skeletal muscle and inhibition of adipogenesis in pre-adipocytes.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Korea University College of Medicine, Seoul 136-701, Korea.

ABSTRACT
Dibenzoylmethane (DBM) has been shown to exert a variety of beneficial effects on human health. However, the mechanism of action is poorly understood. In this study, DBM increased phosphorylation of AMP-activated protein kinase (AMPK) and stimulated glucose uptake in a skeletal muscle cell line. Both knockdown of AMPK with siRNA and inhibition with AMPK inhibitor blocked DBM-induced glucose uptake. DBM increased the concentration of intracellular calcium and glucose uptake due to DBM was abolished by STO-609 (a calcium/calmodulin-dependent protein kinase inhibitor). DBM stimulated phosphorylation of p38 mitogen-activated protein kinase (p38 MAPK), which was blocked by pretreatment with compound C, an AMPK inhibitor. The expression of glucose transporter type 4 (GLUT4) was increased by DBM. The translocation of GLUT4 to the plasma membrane was also increased by DBM in AMPK dependently. In addition, DBM suppressed weight gain and prevented fat accumulation in the liver and abdomen in mice fed a high-fat diet. In pre-adipocyte cells, DBM decreased the activity of acetyl-CoA carboxylase (ACC), the rate-limiting enzyme of fatty acid synthesis. Expression of the adipogenic gene, fatty acid synthase (FAS), was suppressed by DBM in an AMPK-dependent manner. These results showed that the beneficial metabolic effects of DBM might be due to regulation of glucose uptake via AMPK in skeletal muscle and inhibition of adipogenesis in pre-adipocytes.

Show MeSH
Related in: MedlinePlus