Aims
There are many obstacles to overcome in the development of new drugs for metabolic diseases, including efficacy and toxicity problems in later stages of drug development. To overcome these problems and predict efficacy and toxicity in early stages, we constructed a new model of insulin resistance in terms of communication between 3T3‐L1 adipocytes and RAW264.7 macrophages by three‐dimensional (3D) culture.
Results
In this study, results focused on the functional resemblance between 3D co–culture of adipocytes and macrophages and adipose tissue in diabetic mice. The 3D mono–culture preadipocytes showed good cell viability and induced cell differentiation to adipocytes, without cell confluence or cell–cell contact and interaction. The 3D co‐cultured preadipocytes with RAW264.7 macrophages induced greater insulin resistance than two‐dimensional and 3D mono‐cultured adipocytes. Additionally, we demonstrated that 3D co‐culture model had functional metabolic similarity to adipose tissue in diabetic mice. We utilized this 3D co‐culture system to screen PPARγ antagonists that might have potential as therapeutic agents for diabetes as demonstrated by an in vivo assay.
Conclusion
This in vitro 3D co‐culture system could serve as a next‐generation platform to accelerate the development of therapeutics for metabolic diseases.
Fractionation of a 70% ethanolic extract from twigs of Lindera obtusiloba BLUME (Japanese spicebush, Tohaku) yielded fi ve fractions of different polarity. The antifi brotic activity within the chloroform phase was best assessed by an in vitro bioassay using rat hepatic stellate cell (HSC) proliferation and their autocrine transforming growth factor beta (TGF-β) expression as sensitive fi brosis-associated read out. Chromatography of the chloroform extract on Sephadex LH-20 or liquid-liquid extractions yielded a crystalline compound as an active principle, which was identifi ed from NMR and ESI-MS analyses, its melting point, and its optical rotation as (7S,7´R,8R,8´R)-3,4:3´,4´-bis(methylenedioxy)-7,9´:7´,9-diepoxy-lignane [(+)-episesamin]. X-Ray diffraction confi rmed the structure and provided, for the fi rst time, directly its absolute confi guration. (+)-Episesamin blocked proliferation and the profi brotic autocrine TGF-β expression HSC without signifi cant cytotoxicity.
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