title: Proteomic analysis of rosiglitazone and guggulsterone treated 3T3-L1 preadipocytes.
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 3T3-L1 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 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.
authors.affiliation: Drug Target Discovery and Development Division, CSIR-Central Drug Research Institute, Jankipuram Extension, Lucknow, UP, India.
authors.forename: Pooja
authors.initials: P
authors.lastname: Pal
authors.forename: Jitendra K
authors.initials: JK
authors.lastname: Kanaujiya
authors.forename: Savita
authors.initials: S
authors.lastname: Lochab
authors.forename: Shashi B
authors.initials: SB
authors.lastname: Tripathi
authors.forename: Sabyasachi
authors.initials: S
authors.lastname: Sanyal
authors.forename: Gerhard
authors.initials: G
authors.lastname: Behre
authors.forename: Arun K
authors.initials: AK
authors.lastname: Trivedi
mesh_terms: D041721:3T3-L1 Cells / Q000187:drug effects / Q000378:metabolism; D017667:Adipocytes / Q000187:drug effects* / Q000378:metabolism*; D050156:Adipogenesis / Q000187:drug effects; D000818:Animals; D017306:Annexin A2 / Q000235:genetics / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D037483:Galectin 1 / Q000235:genetics / Q000378:metabolism; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D051379:Mice; D011282:Pregnenediones / Q000494:pharmacology*; D040901:Proteomics / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction; D000077154:Rosiglitazone; D045162:Thiazolidinediones / Q000494:pharmacology*; D000090324:Tumor Protein, Translationally-Controlled 1
chemical_list: D017306:Annexin A2; D014408:Biomarkers, Tumor; D037483:Galectin 1; D007004:Hypoglycemic Agents; D011282:Pregnenediones; D045162:Thiazolidinediones; C000715648:Tpt1 protein, mouse; D000090324:Tumor Protein, Translationally-Controlled 1; D000077154:Rosiglitazone; C023617:pregna-4,17-diene-3,16-dione