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The small molecule indirubin-3'-oxime activates Wnt/β-catenin signaling and inhibits adipocyte differentiation and obesity.

Choi OM, Cho YH, Choi S, Lee SH, Seo SH, Kim HY, Han G, Min DS, Park T, Choi KY - Int J Obes (Lond) (2013)

Bottom Line: We investigated the effect of indirubin-3'-oxime (I3O), which was screened as an activator of the Wnt/β-catenin signaling, on inhibiting the preadipocyte differentiation in vitro and in vivo. 3T3L1 preadipocytes were differentiated with 0, 4 or 20 μM of I3O.The I3O effect on adipocyte differentiation was observed by Oil-red-O staining.I3O inhibited the differentiation of 3T3-L1 cells into mature adipocytes and decreased the expression of adipocyte markers, CCAAT/enhancer-binding protein α and peroxisome proliferator-activated receptor γ, at both mRNA and protein levels.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea [2] Translational Research Center for Protein Function Control, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea.

ABSTRACT

Objectives: Activation of the Wnt/β-catenin signaling pathway inhibits adipogenesis by maintaining preadipocytes in an undifferentiated state. We investigated the effect of indirubin-3'-oxime (I3O), which was screened as an activator of the Wnt/β-catenin signaling, on inhibiting the preadipocyte differentiation in vitro and in vivo.

Methods: 3T3L1 preadipocytes were differentiated with 0, 4 or 20 μM of I3O. The I3O effect on adipocyte differentiation was observed by Oil-red-O staining. Activation of Wnt/β-catenin signaling in I3O-treated 3T3L1 cells was shown using immunocytochemical and immunoblotting analyses for β-catenin. The regulation of adipogenic markers was analyzed via real-time reverse transcription-PCR (RT-PCR) and immunoblotting analyses. For the in vivo study, mice were divided into five different dietary groups: chow diet, high-fat diet (HFD), HFD supplemented with I3O at 5, 25 and 100 mg kg(-1). After 8 weeks, adipose and liver tissues were excised from the mice and subject to morphometry, real-time RT-PCR, immunoblotting and histological or immunohistochemical analyses. In addition, adipokine and insulin concentrations in serum of the mice were accessed by enzyme-linked immunosorbent assay.

Results: Using a cell-based approach to screen a library of pharmacologically active small molecules, we identified I3O as a Wnt/β-catenin pathway activator. I3O inhibited the differentiation of 3T3-L1 cells into mature adipocytes and decreased the expression of adipocyte markers, CCAAT/enhancer-binding protein α and peroxisome proliferator-activated receptor γ, at both mRNA and protein levels. In vivo, I3O inhibited the development of obesity in HFD-fed mice by attenuating HFD-induced body weight gain and visceral fat accumulation without showing any significant toxicity. Factors associated with metabolic disorders such as hyperlipidemia and hyperglycemia were also improved by treatment of I3O.

Conclusion: Activation of the Wnt/β-catenin signaling pathway can be used as a therapeutic strategy for the treatment of obesity and metabolic syndrome and implicates I3O as a candidate anti-obesity agent.

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Related in: MedlinePlus

Effects of I3O on HFD-induced obesity in C57BL/6N mice. Fifty male C57BL/6N mice were divided into groups (n=10 per group) and fed a chow diet, HFD, or HFD supplemented with I3O at 5, 25 and 100 mg kg−1. (a) Whole-body images of Representative control and I3O-treated mice. (b) Total body weight. (c) Representative images of visceral white adipose tissue and liver of mice. Plasma levels of triglyceride (d), total cholesterol (e), free fatty acid (f), and glucose levels (g) in mice after 8 weeks of feeding with chow, HFD, or I3O-supplemented HFD. Values are expressed as means±s.e.m. (n=10). *P<0.05, **P<0.01, ***P<0.001 (Kruskal-Wallis test).
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fig3: Effects of I3O on HFD-induced obesity in C57BL/6N mice. Fifty male C57BL/6N mice were divided into groups (n=10 per group) and fed a chow diet, HFD, or HFD supplemented with I3O at 5, 25 and 100 mg kg−1. (a) Whole-body images of Representative control and I3O-treated mice. (b) Total body weight. (c) Representative images of visceral white adipose tissue and liver of mice. Plasma levels of triglyceride (d), total cholesterol (e), free fatty acid (f), and glucose levels (g) in mice after 8 weeks of feeding with chow, HFD, or I3O-supplemented HFD. Values are expressed as means±s.e.m. (n=10). *P<0.05, **P<0.01, ***P<0.001 (Kruskal-Wallis test).

Mentions: Male C57BL/6N mice were fed a HFD supplemented with I3O (Supplementary Figure 5) for 8 weeks. Fifty mice were divided into five groups (n=10 for each group): chow diet, HFD and I3O-supplemented HFD at 5, 25 and 100 mg kg−1 for a day. After 8 weeks, mice fed a HFD had significantly higher body weights and more abdominal fat than mice fed a chow diet (Figures 3a and b). There were no deaths or obvious signs of toxicity during the 8 weeks of feeding. Quantitative analysis of total body weight and weight gain indicated a significant reduction in obesity in the mice fed an I3O-supplemented HFD compared with mice fed a regular HFD. The average final weight of the mice was reduced by 3.3, 30 and 41% in the mice treated with 5, 25 and 100 mg kg−1 I3O, respectively (Figure 3b). The body weight gain was also dose-dependently reduced by treatment of I3O, with a 75% reduction in weight gain in the 100 mg kg−1 I3O group compared with the HFD mice (Supplementary Figure 6A). We also observed a slight reduction in daily food intake in I3O-supplemented HFD mice compared with HFD mice (Supplementary Figure 6B). To determine whether food intake had a significant role in the reduction of body weight gain, the food efficiency ratio was calculated for each group of mice (Supplementary Figure 6C). Based on the food efficiency ratio, we concluded that the food intake did not have a significant role in the reduction of body weight gain by I3O.


The small molecule indirubin-3'-oxime activates Wnt/β-catenin signaling and inhibits adipocyte differentiation and obesity.

Choi OM, Cho YH, Choi S, Lee SH, Seo SH, Kim HY, Han G, Min DS, Park T, Choi KY - Int J Obes (Lond) (2013)

Effects of I3O on HFD-induced obesity in C57BL/6N mice. Fifty male C57BL/6N mice were divided into groups (n=10 per group) and fed a chow diet, HFD, or HFD supplemented with I3O at 5, 25 and 100 mg kg−1. (a) Whole-body images of Representative control and I3O-treated mice. (b) Total body weight. (c) Representative images of visceral white adipose tissue and liver of mice. Plasma levels of triglyceride (d), total cholesterol (e), free fatty acid (f), and glucose levels (g) in mice after 8 weeks of feeding with chow, HFD, or I3O-supplemented HFD. Values are expressed as means±s.e.m. (n=10). *P<0.05, **P<0.01, ***P<0.001 (Kruskal-Wallis test).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4125748&req=5

fig3: Effects of I3O on HFD-induced obesity in C57BL/6N mice. Fifty male C57BL/6N mice were divided into groups (n=10 per group) and fed a chow diet, HFD, or HFD supplemented with I3O at 5, 25 and 100 mg kg−1. (a) Whole-body images of Representative control and I3O-treated mice. (b) Total body weight. (c) Representative images of visceral white adipose tissue and liver of mice. Plasma levels of triglyceride (d), total cholesterol (e), free fatty acid (f), and glucose levels (g) in mice after 8 weeks of feeding with chow, HFD, or I3O-supplemented HFD. Values are expressed as means±s.e.m. (n=10). *P<0.05, **P<0.01, ***P<0.001 (Kruskal-Wallis test).
Mentions: Male C57BL/6N mice were fed a HFD supplemented with I3O (Supplementary Figure 5) for 8 weeks. Fifty mice were divided into five groups (n=10 for each group): chow diet, HFD and I3O-supplemented HFD at 5, 25 and 100 mg kg−1 for a day. After 8 weeks, mice fed a HFD had significantly higher body weights and more abdominal fat than mice fed a chow diet (Figures 3a and b). There were no deaths or obvious signs of toxicity during the 8 weeks of feeding. Quantitative analysis of total body weight and weight gain indicated a significant reduction in obesity in the mice fed an I3O-supplemented HFD compared with mice fed a regular HFD. The average final weight of the mice was reduced by 3.3, 30 and 41% in the mice treated with 5, 25 and 100 mg kg−1 I3O, respectively (Figure 3b). The body weight gain was also dose-dependently reduced by treatment of I3O, with a 75% reduction in weight gain in the 100 mg kg−1 I3O group compared with the HFD mice (Supplementary Figure 6A). We also observed a slight reduction in daily food intake in I3O-supplemented HFD mice compared with HFD mice (Supplementary Figure 6B). To determine whether food intake had a significant role in the reduction of body weight gain, the food efficiency ratio was calculated for each group of mice (Supplementary Figure 6C). Based on the food efficiency ratio, we concluded that the food intake did not have a significant role in the reduction of body weight gain by I3O.

Bottom Line: We investigated the effect of indirubin-3'-oxime (I3O), which was screened as an activator of the Wnt/β-catenin signaling, on inhibiting the preadipocyte differentiation in vitro and in vivo. 3T3L1 preadipocytes were differentiated with 0, 4 or 20 μM of I3O.The I3O effect on adipocyte differentiation was observed by Oil-red-O staining.I3O inhibited the differentiation of 3T3-L1 cells into mature adipocytes and decreased the expression of adipocyte markers, CCAAT/enhancer-binding protein α and peroxisome proliferator-activated receptor γ, at both mRNA and protein levels.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea [2] Translational Research Center for Protein Function Control, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea.

ABSTRACT

Objectives: Activation of the Wnt/β-catenin signaling pathway inhibits adipogenesis by maintaining preadipocytes in an undifferentiated state. We investigated the effect of indirubin-3'-oxime (I3O), which was screened as an activator of the Wnt/β-catenin signaling, on inhibiting the preadipocyte differentiation in vitro and in vivo.

Methods: 3T3L1 preadipocytes were differentiated with 0, 4 or 20 μM of I3O. The I3O effect on adipocyte differentiation was observed by Oil-red-O staining. Activation of Wnt/β-catenin signaling in I3O-treated 3T3L1 cells was shown using immunocytochemical and immunoblotting analyses for β-catenin. The regulation of adipogenic markers was analyzed via real-time reverse transcription-PCR (RT-PCR) and immunoblotting analyses. For the in vivo study, mice were divided into five different dietary groups: chow diet, high-fat diet (HFD), HFD supplemented with I3O at 5, 25 and 100 mg kg(-1). After 8 weeks, adipose and liver tissues were excised from the mice and subject to morphometry, real-time RT-PCR, immunoblotting and histological or immunohistochemical analyses. In addition, adipokine and insulin concentrations in serum of the mice were accessed by enzyme-linked immunosorbent assay.

Results: Using a cell-based approach to screen a library of pharmacologically active small molecules, we identified I3O as a Wnt/β-catenin pathway activator. I3O inhibited the differentiation of 3T3-L1 cells into mature adipocytes and decreased the expression of adipocyte markers, CCAAT/enhancer-binding protein α and peroxisome proliferator-activated receptor γ, at both mRNA and protein levels. In vivo, I3O inhibited the development of obesity in HFD-fed mice by attenuating HFD-induced body weight gain and visceral fat accumulation without showing any significant toxicity. Factors associated with metabolic disorders such as hyperlipidemia and hyperglycemia were also improved by treatment of I3O.

Conclusion: Activation of the Wnt/β-catenin signaling pathway can be used as a therapeutic strategy for the treatment of obesity and metabolic syndrome and implicates I3O as a candidate anti-obesity agent.

Show MeSH
Related in: MedlinePlus