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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) Effects of DBM on body weight in an HFD-induced obesity model.The three groups were as follows: LFD, low-fat (standard) diet; HFD, high-fat diet; and HFD supplemented with 100 mg/kg/d DBM. (B) Effects of DBM on epididymal fat in an HFD-induced obesity model. (C) Effects of DBM on peri-renal fat in an HFD-induced obesity model. (D) Effects of DBM on fatty liver in an HFD-induced obesity model. (E) Effects of DBM on insulin level of HFD-induced animals. * p < 0.05, as compared with basal condition. * + p < 0.05, as compared with HFD condition (F) Effects of DBM on fasting blood glucose (FBG) level of HFD-induced animals. * p < 0.05, as compared with basal condition. (G) Effects of DBM on leptin level of HFD-induced animals. * p < 0.05, as compared with basal condition. (H) 3T3-L1 pre-adipocyte cells were stimulated for indicated times with DBM. The cells were then lysed with lysis buffer, and the phosphorylation of acetyl CoA carboxylase (ACC) was assessed by western blot using antibodies specific for the phosphorylate protein. The levels of ACC were also assessed. The levels of β-actin were also measured as a control for protein loading. * p < 0.05, as compared with basal condition. The results are representative of three independent experiments. (I) Total mRNA was prepared from DBM-treated 3T3-L1 cells, and RT-PCR was conducted using specific indicated primers. The PCR products were then separated on 1% agarose gels and visualized under ultraviolet light. Beta-actin was used as a loading control. * p < 0.05, as compared with basal condition. (J) ST3-L1 cells were transiently transfected with AMPKα2 siRNA for 48 h. The cells were then stimulated with 30 μM DBM for 1 h. Total mRNA was prepared from DBM-treated 3T3-L1 cells and RT-PCR was conducted using specific fatty acid synthase (FAS) primers. The PCR products were then separated on 1% agarose gels and visualized under ultraviolet light. Beta-actin was used as a loading control. * p < 0.05, as compared with basal condition.
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pone.0120104.g006: (A) Effects of DBM on body weight in an HFD-induced obesity model.The three groups were as follows: LFD, low-fat (standard) diet; HFD, high-fat diet; and HFD supplemented with 100 mg/kg/d DBM. (B) Effects of DBM on epididymal fat in an HFD-induced obesity model. (C) Effects of DBM on peri-renal fat in an HFD-induced obesity model. (D) Effects of DBM on fatty liver in an HFD-induced obesity model. (E) Effects of DBM on insulin level of HFD-induced animals. * p < 0.05, as compared with basal condition. * + p < 0.05, as compared with HFD condition (F) Effects of DBM on fasting blood glucose (FBG) level of HFD-induced animals. * p < 0.05, as compared with basal condition. (G) Effects of DBM on leptin level of HFD-induced animals. * p < 0.05, as compared with basal condition. (H) 3T3-L1 pre-adipocyte cells were stimulated for indicated times with DBM. The cells were then lysed with lysis buffer, and the phosphorylation of acetyl CoA carboxylase (ACC) was assessed by western blot using antibodies specific for the phosphorylate protein. The levels of ACC were also assessed. The levels of β-actin were also measured as a control for protein loading. * p < 0.05, as compared with basal condition. The results are representative of three independent experiments. (I) Total mRNA was prepared from DBM-treated 3T3-L1 cells, and RT-PCR was conducted using specific indicated primers. The PCR products were then separated on 1% agarose gels and visualized under ultraviolet light. Beta-actin was used as a loading control. * p < 0.05, as compared with basal condition. (J) ST3-L1 cells were transiently transfected with AMPKα2 siRNA for 48 h. The cells were then stimulated with 30 μM DBM for 1 h. Total mRNA was prepared from DBM-treated 3T3-L1 cells and RT-PCR was conducted using specific fatty acid synthase (FAS) primers. The PCR products were then separated on 1% agarose gels and visualized under ultraviolet light. Beta-actin was used as a loading control. * p < 0.05, as compared with basal condition.

Mentions: To determine whether DBM exerts metabolic effects in vivo, we evaluated its effects on HFD-induced changes in body weight. The administration of DBM suppressed HFD-induced increases in body weight (Fig. 6A). To observe the effect by DBM on lipid deposition, images of abdominal fat were also captured in HFD-fed animals. The amounts of epididymal and peri-renal fat were significantly lower in DBM-fed mice than in HFD-fed mice (Fig. 6B and C). The degree of fat deposition in liver due to the HFD was suppressed by DBM administration (Fig. 6D). The color, swelling, size, and fatty surface area of livers in the DBM treatment group were improved compared to controls. The images of organs of low fat diet-animals were also provided. We also analyzed blood samples of DBM-administrated mice. The increased level of insulin in HFD animals down regulated in DBM-administrated mice (Fig. 6E). Blood glucose concentrations were slightly lower in DBM–fed mice than in HFD-fed mice (Fig. 6F). Leptin level was not significantly changed by DBM administration (Fig. 6G). To explore the anti-adipogenic effect of DBM in vitro, we assessed the phosphorylation of acetyl-CoA carboxylase (ACC), a key regulatory enzyme in fatty acid synthesis, in pre-adipocyte 3T3-L1 cells. The administration of DBM increased ACC phosphorylation (Fig. 6H). Phosphorylation of ACC is indicative of a decrease in ACC activity, suggesting that DBM has anti-adipogenic activity through this mechanism. To gain additional insight into the mechanism of DBM, we used reverse transcription-polymerase chain reaction (RT-PCR) to measure the expression of various adipogenesis-related genes. Among tested genes, FAS expression was suppressed dramatically by DBM treatment (Fig. 6I). The downregulation of FAS due to DBM was not observed in AMPKα2 knockdown conditions (Fig. 6J), indicating that DBM suppresses the adipogenic process through AMPK. These results suggested that DBM played an anti-adipogenic role via AMPK-mediated adipogenic gene suppression.


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) Effects of DBM on body weight in an HFD-induced obesity model.The three groups were as follows: LFD, low-fat (standard) diet; HFD, high-fat diet; and HFD supplemented with 100 mg/kg/d DBM. (B) Effects of DBM on epididymal fat in an HFD-induced obesity model. (C) Effects of DBM on peri-renal fat in an HFD-induced obesity model. (D) Effects of DBM on fatty liver in an HFD-induced obesity model. (E) Effects of DBM on insulin level of HFD-induced animals. * p < 0.05, as compared with basal condition. * + p < 0.05, as compared with HFD condition (F) Effects of DBM on fasting blood glucose (FBG) level of HFD-induced animals. * p < 0.05, as compared with basal condition. (G) Effects of DBM on leptin level of HFD-induced animals. * p < 0.05, as compared with basal condition. (H) 3T3-L1 pre-adipocyte cells were stimulated for indicated times with DBM. The cells were then lysed with lysis buffer, and the phosphorylation of acetyl CoA carboxylase (ACC) was assessed by western blot using antibodies specific for the phosphorylate protein. The levels of ACC were also assessed. The levels of β-actin were also measured as a control for protein loading. * p < 0.05, as compared with basal condition. The results are representative of three independent experiments. (I) Total mRNA was prepared from DBM-treated 3T3-L1 cells, and RT-PCR was conducted using specific indicated primers. The PCR products were then separated on 1% agarose gels and visualized under ultraviolet light. Beta-actin was used as a loading control. * p < 0.05, as compared with basal condition. (J) ST3-L1 cells were transiently transfected with AMPKα2 siRNA for 48 h. The cells were then stimulated with 30 μM DBM for 1 h. Total mRNA was prepared from DBM-treated 3T3-L1 cells and RT-PCR was conducted using specific fatty acid synthase (FAS) primers. The PCR products were then separated on 1% agarose gels and visualized under ultraviolet light. Beta-actin was used as a loading control. * p < 0.05, as compared with basal condition.
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pone.0120104.g006: (A) Effects of DBM on body weight in an HFD-induced obesity model.The three groups were as follows: LFD, low-fat (standard) diet; HFD, high-fat diet; and HFD supplemented with 100 mg/kg/d DBM. (B) Effects of DBM on epididymal fat in an HFD-induced obesity model. (C) Effects of DBM on peri-renal fat in an HFD-induced obesity model. (D) Effects of DBM on fatty liver in an HFD-induced obesity model. (E) Effects of DBM on insulin level of HFD-induced animals. * p < 0.05, as compared with basal condition. * + p < 0.05, as compared with HFD condition (F) Effects of DBM on fasting blood glucose (FBG) level of HFD-induced animals. * p < 0.05, as compared with basal condition. (G) Effects of DBM on leptin level of HFD-induced animals. * p < 0.05, as compared with basal condition. (H) 3T3-L1 pre-adipocyte cells were stimulated for indicated times with DBM. The cells were then lysed with lysis buffer, and the phosphorylation of acetyl CoA carboxylase (ACC) was assessed by western blot using antibodies specific for the phosphorylate protein. The levels of ACC were also assessed. The levels of β-actin were also measured as a control for protein loading. * p < 0.05, as compared with basal condition. The results are representative of three independent experiments. (I) Total mRNA was prepared from DBM-treated 3T3-L1 cells, and RT-PCR was conducted using specific indicated primers. The PCR products were then separated on 1% agarose gels and visualized under ultraviolet light. Beta-actin was used as a loading control. * p < 0.05, as compared with basal condition. (J) ST3-L1 cells were transiently transfected with AMPKα2 siRNA for 48 h. The cells were then stimulated with 30 μM DBM for 1 h. Total mRNA was prepared from DBM-treated 3T3-L1 cells and RT-PCR was conducted using specific fatty acid synthase (FAS) primers. The PCR products were then separated on 1% agarose gels and visualized under ultraviolet light. Beta-actin was used as a loading control. * p < 0.05, as compared with basal condition.
Mentions: To determine whether DBM exerts metabolic effects in vivo, we evaluated its effects on HFD-induced changes in body weight. The administration of DBM suppressed HFD-induced increases in body weight (Fig. 6A). To observe the effect by DBM on lipid deposition, images of abdominal fat were also captured in HFD-fed animals. The amounts of epididymal and peri-renal fat were significantly lower in DBM-fed mice than in HFD-fed mice (Fig. 6B and C). The degree of fat deposition in liver due to the HFD was suppressed by DBM administration (Fig. 6D). The color, swelling, size, and fatty surface area of livers in the DBM treatment group were improved compared to controls. The images of organs of low fat diet-animals were also provided. We also analyzed blood samples of DBM-administrated mice. The increased level of insulin in HFD animals down regulated in DBM-administrated mice (Fig. 6E). Blood glucose concentrations were slightly lower in DBM–fed mice than in HFD-fed mice (Fig. 6F). Leptin level was not significantly changed by DBM administration (Fig. 6G). To explore the anti-adipogenic effect of DBM in vitro, we assessed the phosphorylation of acetyl-CoA carboxylase (ACC), a key regulatory enzyme in fatty acid synthesis, in pre-adipocyte 3T3-L1 cells. The administration of DBM increased ACC phosphorylation (Fig. 6H). Phosphorylation of ACC is indicative of a decrease in ACC activity, suggesting that DBM has anti-adipogenic activity through this mechanism. To gain additional insight into the mechanism of DBM, we used reverse transcription-polymerase chain reaction (RT-PCR) to measure the expression of various adipogenesis-related genes. Among tested genes, FAS expression was suppressed dramatically by DBM treatment (Fig. 6I). The downregulation of FAS due to DBM was not observed in AMPKα2 knockdown conditions (Fig. 6J), indicating that DBM suppresses the adipogenic process through AMPK. These results suggested that DBM played an anti-adipogenic role via AMPK-mediated adipogenic gene suppression.

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