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

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.

Indirubin, a small molecular deriving from connectivity map (CMAP) screening, ameliorates obesity-induced metabolic dysfunction by enhancing brown adipose thermogenesis and white adipose browning

“…It is interesting to notice that different compounds with even modest structure changes may lead to quite different physiological effects. One recent research has suggested that indirubin-3′-oxime (I3O), which is a synthesized analog of indirubin, has potential to prevent obesity and metabolic syndrome by inhibiting the differentiation of preadipocytes into mature adipocytes [86]. In our study, we noted that treatment of brown adipocytes with I3O in vitro can only displayed a slight but no significant increase in the expression of BAT marker gene Ucp1 ( Supplymentary Figure 2A), suggest different mechanism involved in the regulation of UCP1 expression compared with indirubin.…”

A new hope for obesity management: Boron inhibits adipogenesis in progenitor cells through the Wnt/β-catenin pathway

“…Although several FDA-approved anti-obesity drugs, including Orlistat, Belviq, and Qsymia, are currently available, their efficiencies remain insufficient (providing a mean weight loss of 3-10%), and they are associated with severe side effects, including birth defects, somnolence, and paraesthesia [25]. Drug development process is hard due to the complex nature of the obesity which is associated with metabolic problems, genetic abnormalities and lifestyle [26].…”

Shockwaves Suppress Adipocyte Differentiation via Decrease in PPARγ

“…The activated Wnt/β-catenin pathway leads to an increase of β-catenin and a decrease of C/EBPα and PPARγ, the main transcriptional factors in adipogenesis. Therefore, Wnt signaling has been suggested as an attractive target for anti-obesity drugs [65]. Inhibition of PPARγ by an antagonist prevented high fat diet (HFD)-induced obesity and improved glucose and lipid metabolism [66].…”