Proteomic analysis of rosiglitazone and guggulsterone treated 3T3-L1 preadipocytes
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Proteomic analysis of rosiglitazone and guggulsterone treated 3T3-L1 preadipocytes Pooja Pal • Jitendra K. Kanaujiya • Savita Lochab Shashi B. Tripathi • Sabyasachi Sanyal • Gerhard Behre • Arun K. Trivedi
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Received: 21 August 2012 / Accepted: 14 December 2012 / Published online: 30 December 2012 Ó Springer Science+Business Media New York 2012
Abstract Adipogenesis is the differentiation of preadipocytes to adipocytes which is marked by the accumulation of lipid droplets. Adipogenic differentiation of 3T3-L1 cells is achieved by exposing the cells to Insulin, Dexamethasone and IBMX for 5–7 days. Thiazolidinedione drugs, like rosiglitazone are potent insulin sensitizing agents and have been shown to enhance lipid droplet formation in 3T3-L1 cells, a model cell line for preadipocyte differentiation. Guggulsterone is a natural drug extracted from the gum resin of tree Commiphora mukul. Guggulsterone has been shown to inhibit adipogenesis and induce apoptosis in 3T3-L1 cells. In this study we treated the 3T3L1 preadipocytes with rosiglitazone and guggulsterone and assessed the protein expression profile using 2D gel electrophoresis-based proteomics to find out differential target proteins of these drugs. The proteins that were identified upon rosiglitazone treatment generally regulate cell proliferation and/or exhibit anti-inflammatory effect which strengthens its differentiation-inducing property. Guggulsterone treatment resulted in the identification of the apoptosis-inducing proteins to be up regulated which rightly is
Electronic supplementary material The online version of this article (doi:10.1007/s11010-012-1551-0) contains supplementary material, which is available to authorized users. P. Pal J. K. Kanaujiya S. Lochab S. B. Tripathi S. Sanyal A. K. Trivedi (&) Drug Target Discovery and Development Division, CSIR-Central Drug Research Institute, New Campus Sector 10, Jankipuram Extension, Sitapur Road, Lucknow 226021, UP, India e-mail: [email protected] G. Behre Division of Hematology and Oncology, University Hospital of Leipzig, Johannissallee 32A, 04103 Leipzig, Germany
in agreement with the apoptosis-inducing property of guggulsterone in 3T3-L1 cells. Some of the proteins identified in our proteomic screen such as Galectin1, AnnexinA2 & TCTP were further confirmed by Real Time qPCR. Thus, the present study provides a better outlook of proteins being differentially regulated/expressed upon treatment with rosiglitazone and guggulsterone. The detailed study of the differentially expressed proteins identified in this proteomic screen may further provide the better molecular insight into the mode of action of these anti-diabetic drugs rosiglitazone and guggulsterone. Keywords Rosiglitazone Guggulsterone Adipogenesis Proteomics Differentiation
Introduction Adipogenesis is the differentiation of preadipocytes to adipocytes with extensive lipid accumulation. It involves the interplay and expression of a wide array of genes and transcription factors including PPARc and C/EBP family proteins [
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